Genetically modified Lactobacillus and uses thereof

The present invention relates to efficient delivery of anti-infective activity, immunomodulatory factors, or growth-promoting biomolecules directly to the digestive tract of an animal via a live delivery platform. The live delivery platform can be a genetically modified microorganism. Delivery can be accomplished with a Lactobacillus sp which colonizes the gastrointestinal tract. The anti-infective activity can be a bacteriocidal or bacteriostatic peptide, an antibody or fragment thereof which specifically recognizes a pathogen, or a phage, or a lytic peptide from a phage which specifically targets a certain pathogen.

1. 11. An expression cassette comprising a promoter for transcriptional expression and at least one heterologous coding region encoding a biomolecule; wherein the biomolecule is a bacterial peptide, an enzyme, a lysin, or a single chain antibody and wherein: (a) the bacterial peptide is selected from mersacidin-E1 and E-2 molecules comprising sequences of SEQ ID NO: 2 and SEQ ID NO: 4, or Bacillus bacteriocin comprising a sequence selected from SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO:47, and SEQ ID NO: 48; (b) the enzyme is selected from SEQ ID NO: 5 or SEQ ID NO: 6; (c) the lysin is selected from SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 28; or (d) the single chain antibody is directed against pathogenic bacterium Clostridium perfringens, specifically recognizes the bacterial protein C. perfringens alpha toxin or C. perfringens NetB toxin, and selected from SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 34, and SEQ ID NO: 50; wherein the promoter comprises the sequence of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
2. 22. The expression cassette of claim 1, wherein the cassette comprises an origin of replication comprising the sequence of SEQ ID NO: 8.
3. 33. The expression cassette of claim 1, wherein the expression cassette is located on a plasmid or suicide vector or is located on or integrated into a bacterial chromosome.
4. 44. The expression cassette of claim 3, wherein the expression cassette is integrated into a bacterial chromosome and is inserted into a transposase locus an Uracil phosphoribosyl (URP) transferase locus, or a pyrE locus of a bacterial chromosome.
5. 55. The expression cassette of claim 1, wherein the single chain antibody directed against pathogenic bacterium Clostridium perfringens comprises one or more sequence selected from SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 49, and SEQ ID NO: 50.
6. 66. The expression cassette of claim 1, wherein the promoter comprises the sequence of SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.

CROSS-REFERENCE TO RELATED APPLICATIONS
The present application is a National Stage Application claiming priority from co-pending PCT Application No. PCT/US2020/016522 filed Feb. 4, 2020, which in turn claims benefit of priority to U.S. Provisional Application Ser. No. 62/801,307, filed Feb. 5, 2019, all of which are is hereby incorporated by reference in their its entirety.


FIELD OF THE INVENTION
The present invention relates to a live delivery platform, such as a genetically modified bacterium, to deliver preventative or therapeutic anti-infective activity, immunomodulatory factors, or growth-promoting biomolecules directly to the mucosa of an animal in need thereof.
BACKGROUND OF THE INVENTION
Direct fed microbials (DFMs), often also called probiotics, are microorganisms which colonize the gastrointestinal tract of an animal and provide some beneficial effect to that animal. The microorganisms can be bacterial species, for example those from the genera Bacillus, Lactobacillus, Lactococcus, and Enterococcus. The microorganisms can also be yeast or even molds. The microorganisms can be provided to an animal orally or mucosally or, in the case of birds, provided to a fertilized egg, i.e. in ovo.
The beneficial activity provided by a DFM can be the synthesis of vitamins or other nutritional molecules needed for a healthy metabolism of the host animal. A DFM can also protect the host animal from disease, disorders, or clinical symptoms caused by other, pathogenic microorganisms. For example, the DFM may naturally produce factors having inhibitory or cytotoxic activity against certain species of pathogens, such as deleterious or disease-causing bacteria. However, the DFM may not be able to produce such factors in sufficient quantity to reduce infection of the host with the pathogen, or the factors may affect only a limited set of pathogens, leaving the host vulnerable to other pathogens.
Stronger or more broad-based antibiotics can be administered to the host animal, for example orally or parenterally, but these would have a limited duration in the host, and thus may require repeated administration. Oral delivery may also result in the degradation of the antibiotics or failure to deliver the antibiotic to the particular anatomical site where the therapeutic effect is most needed. Development of antibiotic resistance by pathogens is another important concern.
What is needed is a delivery system which can constantly deliver anti-infective molecules directly to the gastrointestinal or respiratory tract where pathogenic bacteria are replicating in the host. The gastrointestinal and respiratory systems are also often a point of entry of the pathogen into the host. Preferably, the delivery system is a live genetically modified microorganism, such as a bacterium, which can colonize the gastrointestinal or respiratory tract of a host and directly deliver antibiotic factors to reduce the number of, or block the entry of, a pathogen. For example, in ovo delivery of a live delivery platform could prevent early colonization of an embryo by pathogens, possibly through competitive exclusion or direct or indirect anti-infective effects. In ovo delivery has the further advantage of bypassing any limitations of colonization by the genetically-modified microorganism due to maternal antibody interference. Preferably, the live bacterial delivery system synthesizes the anti-infective factor in sufficient quantity to have the desired effect on a pathogen. A targeted pathogen may be, without limitation, a bacterium of the genera Salmonella, Clostridium, Campylobacter, Fusobacterium, Staphylococcus, or Streptococcus, or an E. coli bacterium, or a parasite such as an Eimeria species. Preferably, the live bacterial system persists in the host gastrointestinal tract for a period of time. Preferably, the live bacterial delivery system produces a broad-spectrum anti-infective factor or multiple anti-infective factors, such that a variety of pathogens are targeted. Alternatively, a combination of live delivery systems could be administered to a single animal, with genetically modified bacteria producing multiple anti-infective factors, immunomodulatory molecules, or growth-promoting biomolecules, or any combination thereof. Thus, more than one disease state is prevented or reduced, or diseases and syndromes having multiple causes can be effectively treated.
Provided herein is disclosure of anti-infective peptides, including new mersacidin-like peptides, which target multiple bacterial species. Also disclosed are antibodies, including single chain antibodies, which target specific pathogens and pathogenic molecules. Also disclosed are phage or phage lytic peptides which target pathogenic species. Provided also is a Lactobacillus expression system which can produce high levels of at least one or a multiplicity of the above molecules, preferably as surface-displayed or secreted molecules.
SUMMARY OF THE INVENTION
The present invention provides a live delivery platform comprising a genetically modified microorganism. The genetically modified microorganism comprises an expression cassette containing one or more of: a promoter for transcriptional expression, a nucleic acid sequence encoding a signal sequence for secretion, a nucleic acid sequence encoding a cell-wall anchor, at least one heterologous coding region encoding a desired biomolecule, a nucleic acid sequence encoding an expressed peptide tag for detection, and terminators for translation and transcription termination. The genetically modified microorganism may be a bacterium, a yeast, or a fungus. A genetically modified bacterium is preferably a Bacillus, Lactobacillus, Lactococcus, or an Enterococcus. The genetically modified bacterium may also preferably be an E. coli bacterium. The genetically modified bacterium may also preferably be a Lactobacillus reuteri strain. The present invention provides an expression cassette within a genetically modified microorganism that may include a promoter for transcriptional expression. The promoter for transcriptional expression may comprise a nucleic acid sequence of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43.
The present invention provides an expression cassette within a genetically modified microorganism that may include a nucleic acid sequence encoding a signal sequence for secretion. The signal sequence for secretion may be at least 20 amino acids, at least 25 amino acids, at least 30 amino acids, at least 35 amino acids, at least 40 amino acids, at least 44 amino acids, at least 50 amino acids, at least 55 amino acids, at least 60 amino acids, or at least 65 amino acids. The signal sequence for secretion may also be 20-65 amino acids, 20-60 amino acids; 20-55 amino acids; 20-50 amino acids, 20-45 amino acids, 20-40 amino acids, 20-35 amino acids, 20-30 amino acids, 25-65 amino acids, 25-60 amino acids; 25-55 amino acids; 25-50 amino acids, 25-45 amino acids, 25-40 amino acids, 25-35 amino acids, 25-30 amino acids, 30-65 amino acids, 30-60 amino acids; 30-55 amino acids; 30-50 amino acids, 30-45 amino acids, 30-40 amino acids, or 30-35 amino acids. The signal sequence for secretion may comprise an amino acid sequence that is a fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein. The signal sequence for secretion may comprise an amino acid sequence that is an amino-terminal fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein.
The present invention provides an expression cassette within a genetically modified microorganism that may include a nucleic acid sequence encoding a cell wall anchor peptide. The cell wall anchor peptide may have a length of 100-250 amino acids, 100-225 amino acids, 100-200 amino acids, 100-175 amino acids, 100-150 amino acids, 125-250 amino acids, 125-225 amino acids, 125-200 amino acids, 125-175 amino acids, or 125-150 amino acids. The cell wall anchor may comprise an amino acid sequence that is a fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein. The cell wall anchor may comprise an amino acid sequence that is a carboxy-terminal fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein.
The present invention provides an expression cassette within a genetically modified microorganism that includes a heterologous coding region encoding a desired biomolecule. The desired biomolecule may be a biomolecule having anti-infective activity, a probiotic factor, an immunomodulatory factor, or a growth-promoting biomolecule. The biomolecule may have anti-infective activity active against a pathogenic bacterium or a parasite. The parasite may preferably be an Eimeria species. The pathogenic bacterium may preferably be a Salmonella, Clostridium, Campylobacter, Staphylococcus, Streptococcus, Fusobacterium, and an E. coli bacterium.
The present invention provides an expression cassette within a genetically modified microorganism that includes a heterologous coding region encoding a desired biomolecule having anti-infective activity. The anti-infective biomolecule may be a bactericidal peptide, an enzyme, a lysin, a phage, or an antibody. The bactericidal peptide may be a mersacidin-like molecule. The mersacidin-like molecule may comprise a sequence disclosed herein as SEQ ID NO: 2 or SEQ ID NO: 4. The desired biomolecule may be an enzyme. The enzyme may comprise a sequence disclosed herein as SEQ ID NO: 5 or SEQ ID NO: 6. The desired biomolecule may be a lysin. The lysin may comprise a sequence disclosed herein as SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28. The desired biomolecule may be a phage, or a phage in a pro-phage form. The phage genetic material may comprise a sequence disclosed herein as SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13. The desired biomolecule may be derived from a Bacillus species. The desired biomolecule may be a Bacillus bacteriocin. A Bacillus anti-infective molecule may comprise a sequence disclosed herein as SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, or SEQ ID NO: 48.
The present invention provides an expression cassette within a genetically modified microorganism that includes a heterologous coding region encoding a desired biomolecule that is an antibody. The antibody may be a single chain antibody. The single chain antibody may be from a camelid. The single chain antibody may specifically recognize a pathogenic microorganism or a molecule produced by a pathogen. The single chain antibody may specifically recognize a bacterial protein, such as for example a toxin or an attachment molecule. The single chain antibody may specifically recognize a bacterial protein from Clostridium perfringens. The bacterial protein may be a protein produced by C. perfringens. The bacterial protein may be C. perfringens alpha toxin or C. perfringens NetB toxin.
The present invention provides an expression cassette within a genetically modified microorganism that includes a heterologous coding region encoding a desired biomolecule that is an antibody that recognizes C. perfringens alpha toxin or C. perfringens NetB toxin. The single chain antibody may comprise an amino acid sequence as disclosed herein as SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, or SEQ ID NO: 34.
The present invention provides an expression cassette within a genetically modified microorganism that includes a heterologous coding region encoding a desired biomolecule that is a probiotic factor. The probiotic factor may be an agglutinin receptor. The agglutinin receptor may comprise a sequence disclosed herein as SEQ ID NO: 9 or SEQ ID NO: 10.
The present invention provides a genetically modified microorganism comprising an expression cassette. The expression cassette may comprise a promoter for transcriptional expression, and at least one heterologous coding region encoding a desired biomolecule. The expression cassette may also optionally comprise one or more of a nucleic acid sequence encoding a signal sequence for secretion, a nucleic acid sequence encoding a cell-wall anchor, a nucleic acid sequence encoding an expressed peptide tag for detection, and terminators for translation and transcription termination. The expression cassette may comprise a promoter having a sequence disclosed herein as SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 41, SEQ ID NO: 42, or SEQ ID NO: 43. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule such as a bactericidal peptide, an enzyme, a lysin, a phage, and an antibody. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having anti-infective activity. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having a sequence disclosed herein as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO:34, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, or SEQ ID NO: 50.
The present invention provides a genetically modified microorganism comprising an expression cassette, where the genetically modified bacterium is a Bacillus, Lactobacillus, Lactococcus, or an Enterococcus. The genetically modified bacterium may be a Lactobacillus reuteri strain.
The present invention provides a genetically modified microorganism comprising an expression cassette, where the expression cassette is located on a plasmid. The plasmid may comprise a sequence disclosed herein as SEQ ID NO: 8. The present invention provides a genetically-modified microorganism comprising an expression cassette, where the expression cassette is located on a bacterial chromosome. The expression cassette located on a bacterial chromosome may be inserted into a transposase locus. The expression cassette located on a bacterial chromosome may be inserted into an Uracil phosphoribosyl (UPP) transferase locus. The expression cassette located on a bacterial chromosome may be inserted into a pyrE locus.
The present invention provides a method of reducing colonization of an animal by a pathogenic bacterium. The method may comprise treating an animal in need thereof with a live delivery platform. The live delivery platform comprises a genetically modified microorganism. The genetically modified microorganism comprises an expression cassette containing one or more of: a promoter for transcriptional expression, a nucleic acid sequence encoding a signal sequence for secretion, a nucleic acid sequence encoding a cell-wall anchor, at least one heterologous coding region encoding a desired biomolecule, a nucleic acid sequence encoding an expressed peptide tag for detection, and terminators for translation and transcription termination. The genetically modified microorganism may be a bacterium, a yeast, or a fungus. A genetically modified bacterium is preferably a Bacillus, Lactobacillus, Lactococcus, or an Enterococcus. The genetically modified bacterium may also preferably be an E. coli bacterium. The genetically modified bacterium may also preferably be a Lactobacillus reuteri strain.
The present invention provides a method of reducing colonization of an animal by a pathogenic bacterium, where the method comprises treating an animal in need thereof with a genetically modified microorganism. The genetically modified microorganism is modified to contain an expression cassette. The expression cassette may comprise a promoter for transcriptional expression having a sequence disclosed herein as SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 41, SEQ ID NO: 42, or SEQ ID NO: 43. The expression cassette may further comprise a signal sequence for secretion having an amino acid sequence that is a fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein. A signal sequence for secretion may comprise an amino acid sequence that is an amino-terminal fragment of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 40, where the fragment has a length as given herein. The expression cassette preferably also includes a heterologous coding region encoding a desired biomolecule. The desired biomolecule may be a biomolecule having anti-infective activity, a probiotic factor, an immunomodulatory factor, or a growth-promoting biomolecule. The biomolecule may have anti-infective activity active against a pathogenic bacterium or a parasite. The parasite may preferably be an Eimeria species. The pathogenic bacterium may preferably be a Salmonella, Clostridium, Campylobacter, Staphylococcus, Streptococcus, Fusobacterium, and an E. coli bacterium. The anti-infective biomolecule may be a bactericidal peptide, an enzyme, a lysin, a phage, or an antibody. The expression cassette may optionally contain a nucleic acid sequence encoding a cell-wall anchor, a nucleic acid sequence encoding an expressed peptide tag for detection, and/or terminators for translation and transcription termination.
The present invention provides a method of reducing colonization of an animal by a pathogenic bacterium, where the method comprises treating an animal in need thereof with a genetically modified microorganism. The animal may be a bird, a human, or a non-human mammal. The treatment may be administered orally, parentally, nasally, or mucosally. When the animal is a bird the treatment may be administered in ovo.
The present invention provides a use of any genetically modified microorganism disclosed herein in therapy. The present invention provides a use in therapy of a bacterium genetically modified to contain any expression cassette as disclosed herein. A therapy may be reducing colonization of an animal by a pathogenic bacterium.
The present invention provides a use of any expression cassette disclosed herein in therapy. The present invention provides a use in therapy of any expression cassette as disclosed herein. A therapy may be reducing colonization of an animal by a pathogenic bacterium. The expression cassette should comprise at least one heterologous coding region encoding a desired biomolecule. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule such as a bactericidal peptide, an enzyme, a lysin, a phage, and an antibody. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having anti-infective activity. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having a sequence disclosed herein as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO:34, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, or SEQ ID NO: 50.
The present invention provides a use of any genetically modified microorganism disclosed herein in the manufacture of a medicament for reducing colonization of an animal by a pathogenic bacterium. The present invention provides for use of a bacterium genetically modified to contain any expression cassette as disclosed herein in the manufacture of a medicament for reducing colonization of an animal by a pathogenic bacterium.
The present invention provides a use of any expression cassette disclosed herein in the manufacture of a medicament for reducing colonization of an animal by a pathogenic bacterium. The present invention provides a use in manufacture of a medicament of any expression cassette as disclosed herein. The medicament may be for reducing colonization of an animal by a pathogenic bacterium. The expression cassette should comprise at least one heterologous coding region encoding a desired biomolecule. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule such as a bactericidal peptide, an enzyme, a lysin, a phage, and an antibody. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having anti-infective activity. The at least one heterologous coding region encoding a desired biomolecule may encode a biomolecule having a sequence disclosed herein as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO:34, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, or SEQ ID NO: 50.
The present invention provides an antibody comprising an amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 50 or SEQ ID NO: 34. The antibody preferably binds a toxin produced by Clostridium perfringens. The antibody preferably binds C. perfringens alpha toxin or a C. perfringens NetB toxin. The present invention provides use of an antibody comprising an amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 50 or SEQ ID NO: 34 in therapy. The present invention provides use of an antibody comprising an amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 50 or SEQ ID NO: 34 in reducing colonization of an animal by a C. perfringens bacterium. The present invention provides use of an antibody comprising an amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 50 or SEQ ID NO: 34 in the manufacture of medicament to reduce colonization of an animal by a C. perfringens bacterium. The present invention provides a method of treating an animal for C. perfringens infection or colonization, where the method comprises administering an antibody comprising an amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 50 or SEQ ID NO: 34 to an animal in need thereof.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1. Antibiotic resistance or sensitivity of Lactobacillus reuteri strains, as determined with an AVIPRO® Plate.
FIG. 2. Pathogenic bacteria, but not nonpathogenic Lactobacillus acidophilus are killed by L. reuteri strain 3632, as evidenced by a clear “halo” surrounding strain 3632.
FIG. 3A and 3B. Chromosomal integration of an expression cassette. A. A non-limiting example of a suicide vector map with a representative expression cassette. B. A map of possible chromosomal integration sites.



DETAILED DESCRIPTION OF THE INVENTION
As used herein, a “genetically-modified microorganism” means any microorganism which has been altered from the natural state using molecular biological techniques. A genetic modification could be the deletion of a portion of the bacterial chromosome or a naturally-occurring plasmid. The genetic modification could also be the introduction of an artificial or exogenous nucleic acid into a portion of the chromosome. The introduction may or may not disturb or perturb the expression of a bacterial gene. The genetic modification could also be the introduction of an artificial plasmid. The genetically-modified microorganism may be a bacterium, a virus, a yeast, a mold, or a single-celled organism.
An “artificial nucleic acid” or “artificial plasmid” is any nucleic acid or plasmid which does not occur naturally, but rather has been constructed using molecular biological techniques. Portions of the nucleic acid or plasmid may occur naturally, but those portions are in an artificial relationship or organization.
As used herein, an “expression cassette” is an artificial nucleic acid constructed to result in the expression of a desired biomolecule by the genetically-modified microorganism. An expression cassette comprises one or more of a promoter for transcriptional expression, a nucleic acid sequence encoding a signal sequence for secretion, a nucleic acid sequence encoding a cell-wall anchor, at least one heterologous coding region encoding a desired biomolecule, a nucleic acid sequence encoding an expressed peptide tag for detection, and terminators for translation and transcription termination. A promoter directs the initiation of transcription of the coding regions into a messenger RNA and the translation of the mRNA into a peptide. A signal sequence for secretion, or a secretion signal sequence, directs the peptide to be located outside the cell membrane. The extracellular peptide could be a soluble, secreted protein or it may be cell-associated, particularly if the expression cassette contains a cell wall anchor sequence which attaches the extracellular peptide to a bacterial cell wall. An expressed peptide tag is any amino acid sequence which may be recognized by an antibody or other binding protein. The expressed peptide tag may also bind an inorganic substance, such as a six-histidine tag which binds to nickel molecules. Terminators for translation may be a stop codon or a spacer open reading frame containing a stop codon.
As used herein, a “heterologous coding region” is a nucleic acid sequence containing an open reading frame which encodes a peptide. The coding region is heterologous to the associated promoter, meaning the coding region and the promoter are not associated in their natural states.
As used herein, a “protein” is a sequence of amino acids which assumes a three-dimensional structure. A “peptide” can be used interchangeably with protein but may also be a short linear sequence of amino acids without a defined three-dimensional structure.
As used herein, a “desired biomolecule” is any peptide which may be advantageous to a host when administered via a live delivery platform. The desired biomolecule may be a peptide with anti-infective activity, a probiotic factor, an immunomodulatory factor, an anti-antinutritional factor, or a growth-promoting biomolecule. The desired biomolecule may also be an enzyme which produces a substance with anti-infective activity or a probiotic factor such as a vitamin.
As used herein, “anti-infective activity” includes any activity which prevents infection of a host with a pathogenic organism. The following molecules are examples of biomolecules possessing anti-infective activity: an antibacterial peptide; a lysin or lytic enzyme; a prophage, phage or virus; an enzyme, for example one that cleaves or disables a protein made by a pathogen; and an antibody which blocks, inhibits, or clears a pathogenic molecule. An anti-infective may have bacteriostatic activity, which slows, reduces, or prevents the growth of a pathogenic species. A non-limiting example of an antibacterial peptide is a member of the mersacidin family or a mersacidin-like molecule, such as those described in EP0700998. A non-limiting example of lysins are lytic molecules produced by phage. Lysins may have specificity for certain pathogenic species of bacteria and have been suggested for use in substitution for traditional antibiotics. V. A. Fischetti, Viruses, vol. 10, no. 310 (2018); and R. Vazquez et al. Frontiers in Immunology, vol. 9, article 2252 (2018).
As used herein, a “probiotic factor” is a substance which, when produced by a genetically-modified microorganism, proves beneficial to a host. The probiotic factor may be an attachment molecule or an agglutinizing molecule which promotes colonization of the host with the genetically modified microorganism and/or prolongs the period of time where the genetically modified microorganism colonizes the host. The longer the genetically-modified microorganism persists in the host the longer the beneficial effect is provided.
As used herein, an “immunomodulatory factor” could be a cytokine, lymphokine, chemokine, interleukin, interferon, a colony stimulating factor, or a growth factor. The immunomodulatory factor could provide nonspecific enhancement of an immune response or the immunomodulatory factor could increase the number or tissue distribution of immune cells present in the host. The immunomodulatory factor may also reduce an inappropriate immune response, such as without limitation an autoimmune response.
As used herein, a “growth-promoting biomolecule” could be a growth factor, a transfer factor (such as an iron-chelating molecule), a hormone, or any other factor which promotes healthy metabolic activity.
As used herein, an “anti-nutritional factor” could include protease inhibitors, for example a trypsin inhibitor.
As used herein, “delivery” or “administration” means the act of providing a beneficial activity to a host. The delivery may be direct or indirect. An administration could be by an oral, nasal, or mucosal route. For example without limitation, an oral route may be an administration through drinking water, a nasal route of administration may be through a spray or vapor, and a mucosal route of administration may be through direct contact with mucosal tissue. Mucosal tissue is a membrane rich in mucous glands such as those that line the inside surface of the nose, mouth, esophagus, trachea, lungs, stomach, gut, intestines, and anus. In the case of birds, administration may be in ovo, i.e. administration to a fertilized egg. In ovo administration can be via a liquid which is sprayed onto the egg shell surface, or an injected through the shell.
As used herein, the terms “treating”, “to treat”, or “treatment”, include restraining, slowing, stopping, reducing, ameliorating, or reversing the progression or severity of an existing symptom, disorder, condition, or disease. A treatment may also be applied prophylactically to prevent or reduce the incidence, occurrence, risk, or severity of a clinical symptom, disorder, condition, or disease. As used herein, the term “reducing” may apply to both prophylactic (e.g. preventative) treatments or therapeutic treatments.
The following experimental examples are illustrative of a live delivery system comprising Lactobacillus expression cassettes which can be delivered by the disclosed live delivery platform. It will be appreciated that other embodiments and uses will be apparent to those skilled in the art and that the invention is not limited to these specific illustrative examples or preferred embodiments. The descriptive headings of these Examples are for convenience only and should not influence interpretation of any of the results presented therein.
Example 1. L. reuteri Strain Identification
Seven Lactobacillus reuteri (L. reuteri) strains are isolated from older birds at the Research Center, Hannover from the cecal contents received from the Poultry Clinic, University of Hannover. All the seven strains are identified to be L. reuteri by 16S rRNA sequencing.
While only limited growth is observed for most strains under aerobic conditions in MRS broth and agar (de Man, J. D.; Rogosa, M.; and Sharpe, M. E. “A Medium for the Cultivation of Lactobacilli”. J. Appl Bact. 23: 130-135 (1960)), all isolates show very good growth on MRS agar and MRS broth under anaerobic conditions at 39° C. Culturing the bacterial strains on blood agar under anaerobic conditions results mostly in limited growth. None of the strains is able to grow in Mueller Hinton broth under anaerobic conditions. For all further analysis, bacterial strains are grown in MRS medium under anaerobic conditions at 39° C.
Antimicrobial susceptibility of bacterial isolates is tested using the AVIPRO® PLATE (FIG. 1). All strains are resistant against colistin, doxycycline, enrofloxacin, erythromycin, neomycin, oxacillin, penicillin G, trimethroprim-sulfamethoxazole, tetracycline, tilmicosin and tylosin. All strains are resistant to streptomycin except strain 3632, and to tiamulin except strain 2098. In addition, resistance to cefpodaxime-proxetil is observed with strains 2091, 2095, 2097 and 3630; resistance to cefotaxime is observed with strains 2091, 2095 and 2097; and resistance to lincomycin is observed with strains 3630 and 3632. No strain is found to be resistant against amoxicillin, ceftiofur, erythromycin D, lincomycin-spectinomycin and rifampicin under tested concentrations. Lactobacillus reuteri strain 3632 was deposited on 19 Jun. 2020 according to the Budapest Treaty in the American Type Culture Collection (ATCC), ATCC Patent Depository, 10801 University Boulevard, Manassas, Va., 20110, USA. The deposit was assigned ATCC Patent Deposit Number PTA-126788. Lactobacillus reuteri strain 3630was deposited on 19 Jun. 2020 according to the Budapest Treaty in the American Type Culture Collection (ATCC), ATCC Patent Depository, 10801 University Boulevard, Manassas, Va., 20110, USA. The deposit was assigned ATCC Patent Deposit Number PTA-126787.
Example 2. L. reuteri Strain Selection and Anti-Infective Activity
To select the best strain for further engineering, the L. reuteri isolates are tested for various desirable probiotic anti-infective properties, such as growth kinetics, ability to produce hydrogen peroxide, auto-aggregation, enzyme profile, survival in the presence of ox bile and pancreatic enzymes, and sensitivity to heat shock and pH changes. The L. reuteri strains are also tested for safety using a haemolytic assay.
In general, all strains behave very similar in terms of probiotic properties, including growth kinetics and ability to produce hydrogen peroxide, except for strain 3632, which shows some unique properties, including the ability to auto-aggregate in liquid media (comparable to that of the well-characterized human probiotic strain L. reuteri ATCC 23272). None of the strains is found to be hemolytic on blood agar plates, suggesting that these isolates are less likely to be pathogenic to humans.
Whole-genome sequencing is performed for L. reuteri strains 2091 and 3632, and an independently isolated L. reuteri strain 170331 of European origin, using PACBIO® sequencing (Amplicon Express). Sequencing, assembly and annotation statistics are summarized in TABLE 1. Genomic structures and organization differ among the tested strains.







TABLE 1







Genome sequencing, assembly and annotation statistics.












Strain
170331
2091
3632







Genome size
2,090,596 bp
2,231,245 bp
2,482,713 bp



Contigs
9
11
7



Coding Sequences
2,154
2,280
2,595



(ORFs)






Ribosomal
2,231
2.354
2,680



Binding Sites






Transcription
1,123
1,123
1,375



Terminators






Operons
452
472
541



tRNA
75
61
77



rRNA
18
19
13



Other RNAs
39
99
89



CRISPR
1
0
0



Prophages
7
1
8



Islands
30
22
24



Bacteriocins
0
1
4










Based on the genome sequencing data, strain 3632 encodes for two bacteriocins belonging to mersacidin family based on homology to the mersacidin conserved domain. These bacteriocins appear to be unique to strain 3632. A cDNA encoding one mersacidin (mersacidin-E1) could be:








(SEQ ID NO: 1)










  1
atggacaaag aagaattaga aaaaattgta ggtaataact ttgaggaaat gagtttacaa



 


 61
aaaatgacag aaattcaagg tatgggtgaa taccaagtgg attcaacacc agcagcttct


 


121
gcgatttcac gggcaacaat tcaagtatca cgtgcatctt ctggaaaatg tctaagttgg


 


181
ggtagtggtg cagcatttag tgcttatttt actcataaaa gatggtgcta g.






(SEQ ID NO: 1). This novel open reading frame would encode a polypeptide of mersacidin-E1:








(SEQ ID NO: 2)


MDKEELEKIVGNNFEEMSLQKMTEIQGMGEYQVDSTPAASAISRATIQVS


 


RASSGKCLSWGSGAAFSAYFTHKRWC.






Another cDNA encoding the second form of mersacidin (mersacidin-E2) could be:








(SEQ ID NO: 3)










  1
atggaagaaa aagaattaga aggtgtaata gggaattcgt ttgaaagtat gactgtagag



 


 61
gaaatgacaa aaattcaagg tatgggtgaa tatcaagtag attcgacgcc tggatatttt


 


121
atggaaagtg ctgccttttc agctcttaca gccaatataa caagacatgc tatgcatcat


 


181
cattaa.






This novel open reading frame would encode a polypeptide of mersacidin-E2:








(SEQ ID NO: 4)


MEEKELEGVIGNSFESMTVEEMTKIQGMGEYQVDSTPGYFMESAAFSALT


 


ANITRHAMHHH.






L. reuteri strain 3632 encodes other polypeptides which contribute anti-infective activity to this strain. Capreomycidine synthase, involved in capreomycin synthesis, is identified to be expressed in both the culture supernatant and pelleted cells.







(SEQ ID NO: 5)


MVEIAHFGVEAWLNKWEKSATYDISQSTIASLSMHDLLNLDGNNGEEFYE


 


MLDKQQMNYGWIEGSPEFKEEVAKLYHHVDPENILQTNGATGANILALYA


 


LINPGDHVIAEYPSYQQLYDIPKSLGADVDYWHIHEEDNWYPRIDDLKAM


 


VKPNTKMICLNNANNPTGTVLDKEFLEQVVEIAKSVDAYVLVDEVYLPLD


 


HPEKFAQIIDLYDKGISTNSLSKTYSVPGVRIGWTATNAEVADIFRKFRD


 


YTMICGGVFNDQLATYVLRHRDQVLARNRKLVLGNLAIYKDWIDHEDRAS


 


VIMPQAVSTSFPKLDVPVDIHTFCENLLHDEGVLLVPGDAFDTPGHVRLG


 


YCAPEATLKEGLKRLSKYMHQYD.






Colicin V production protein is also expressed in pelleted cells of strain 3632:







(SEQ ID NO: 6)


MILTTFIILILMGCFINGHRRGLLTMTLMLGTYIVAWIVARQGAQLIGG


 


WLKSLLPSIGTPATFSESLLANVNSNLFFYNGIAFMIIFTIVSILCHWG


 


IRQLNWIKRIPVVGTVDKIAGGLISFLIGYLITYVVLLIMQLFPAGWWQ


 


MQIANSELARFMINQTPGIAHLVIDTLVQGG.






A person of skill in the art would recognize that, because of the redundancy of the genetic code, multiple nucleic acid sequences could encode the above peptides.

The functionality of these putative bacteriocins is tested by co-plating pathogenic bacteria with strain 3632. As shown in FIG. 2, the pathogenic bacteria Clostridium perfringens, Staphylococcus aureus, Streptococcus suis, Streptococcus uberis, Escherichia coli (APEC), Salmonella typhimurium, Fusobacterium necrophorum, and Campylobacter jejuni are killed by L. reuteri 3632. A non-pathogenic control, Lactobacillus acidophilus, is not killed. DFM strains such as 3632 may be used to control, limit, prevent, or eliminate infection by pathogenic bacteria in swine (S. suis), poultry (C. perfringens, E. coli, S. typhimurium, and C. jejuni), and cattle (S. aureus, S. uberis, and F. necrophorum). DFM strains such as 3632 may be used to control, limit, prevent, or eliminate infection by food-borne pathogenic bacteria in humans.
Example 3. Other Genetic Elements of L. reuteri Strain 3632
In order to identify any potential plasmids containing antimicrobial resistance genes, native plasmids are isolated and identified by high throughput sequencing (ACGT, Inc). The 3632 strain contains three native plasmids but none with any known antimicrobial resistance genes. However, one of the plasmids is maintained in high copy numbers:








(SEQ ID NO: 7)










1
TTGGCAATCT TTCACCTATC AGCAAAAATC ATTAGTCGAG GAAAAGGGCA ATCAGCAATT



 


61
GCCTCGGCTG CTTATCGTTC AGGGAACAAA CTTCACGATG AACGATACGA CGAAACACAA


 


121
GATTACACAA ACAAACGTTT CATCGAACAC TCAGAAATCC AACTTCCAGA GAATGCCCCA


 


181
GCTAAGTATC AAGATAGGGC AACCCTTTGG AACAGCGTAG AAAAGGCAGA AAAAGCTAAG


 


241
AATTCGCAGT TAGCTAGAGA AATTGAAATA GCCTTACCAC GAGAGCTAAC ACCAGAACAA


 


301
CGAGTTAAAT TAGTCCACGA TTACGTTCAG AAAACCTTTG TCGATAAGGG AATGGTAGCC


 


361
GATTGGTCTA TTCACAACCC ACAACCAGAT AAGGATAATC CAGAAAAGCC AGCAAACCCG


 


421
CACGCTCATA TCATGCTCAC ACTGCGTAGC TTGCGTTCTA ACGGCTCTTG GGCACCAAAG


 


481
AAGACAAGTC ACTATGAACT AGACGAAAAC GGCCAGAAGG TGCCTGTAAT CGATCCTGAG


 


541
ACAGGCAAAC AGAAGTTAGG GGCGAAAAAC CAAAAAATCT GGAAACGAGT AATTACCCCA


 


601
ACTAATGACT GGAATAACCC CAAAAACGTT GAAAAATGGC GGGCTGAATG GGCGAAGACT


 


661
TGTAATAAGT ACTTGGCTCC TGACCACCAG ATTGACCACC GCAGTTATAA ACGGCAGGGC


 


721
AAAAAACAAA TTCCAACAAT CCATGAGGGC TATGTTGCTC GTAAAATGGA ACGAGAAGCC


 


781
ACAGGAAGCT CAGAGAGGGC TTCTTTTAAC CAAATCGTAA AATATATCAA CAACGAGTTA


 


841
AAATCGCTTA GAAAGCAAAT TAGGGGCATT ATACGCGAAA TACGGAATCT AGAAAAAGGA


 


901
CGTGACCAAG ATGAGAAAGT TCGACTTCAG CAAGACACCC GAGCAAATCA AACAAGAACA


 


961
AGCAGAGATC AAGCGCTGGG AGGAGGAGTT CAACGCACGA TTACAGACAA TCGGGAGCAA


 


1021
TCCGGAAACG GTAGAGCCAA CGGAACTGCC GAAGAAGCCC AACGAACAGA TCCTAATCAA


 


1081
CTTCATCAAC AGTCTTTCGG AGAAACAGCA AGAAGAATTT TCCGAGATTT CCGGCTATTT


 


1141
GCGGAGCGTC AACGAGAAGC TCGACAGCGA CAACGCCGAC TTAGCCAAGG CTTACAACAT


 


1201
CATCAAGAAA CAGAAAAACG ACTTGAAGCA GGGCAACAAC AACTTACTCG AACAAAACAT


 


1261
ACAGCTGACG AACGAAAACA ACAGCTTACA GAACGACAAA TTCGAGTTAG AAAAGCTTTT


 


1321
AGCGACGCTT CTAAGTCCAA AGTCTTGGAA CGATACTACA CTCACTTCCG TCAAACAAAA


 


1381
ACTCAAGCAA ATCATGCAAA AGGATTAGAC TACCAAGGAC CAATTCTGGG ACATTCCCGA


 


1441
GGACGCTCAA GGTAGTTATG CTAGAATGAT TAAGCCGTCC GTGGCAACTC CTACGTAAAG


 


1501
GAGGTGGCAA CATGGCACAT ATACTCCTCT CAGCAGTTTT GAGCCTTCTG GGTTCCTTGA


 


1561
TAACTGCACT GTTCGCTGAT TGGCTTCGTC GGCGAAAATA GTTTTTGACG GCATGAAGCG


 


1621
CAAATTAAAA GCCACTCAAC TATTCGCACT AGTTGGGTGG CTTTTTTGCA ACGTGGCAAT


 


1681
ATACTTCTCT CGGTTAAATT ATAACATGGT TGTTCCAGAA GTGCATTAGG TAGGCGGTTT


 


1741
GTCCCCCCCC ATGGGTTTTG GCTTCGAAAA TCAATCGTTG AAGTGACCAG CGAAGCTCAG


 


1801
CTGATGGAGC CCTCATATTC AAGAAGCAGG TCAACGAATG CCAATTCCAA CATGAGGGCC


 


1861
ATGACTTAGG CCACTGATAC TCGCAAATGA CTGTCACACC AACGGTATTA GCCATTCTGA


 


1921
TAAGCGAGTC TAACGACTTT TGACGCATAA TCGACCACCT ACCTTATTTT CAAAGCGCCT


 


1981
CAGAGCTAAT TAGAGTATTC TTCAAGGAGC TTAGCGAGGA TTATAGACGC TTGGCGGACT


 


2041
TCACTTTCGG CTTGATTGAC CCGTTTAGCG ATCTTGTCAT CATGGCGAGC CACCTTTTCC


 


2101
AAGTCATGGA GATTATCAGC GATTTGGTCA GTCATGACGG AGTAGTTCTT AATACCCGAC


 


2161
AGCAGGCTTT CAACGTTAGT ATTCTCAAGG TTTTCGTTGA ATATCATGGT TATTCCTCCT


 


2221
TATCCTCAAA CTTAGTCATC TTCTGTCTCC TGTTTTAGAG AATTTTCAAT CATGTCTAAT


 


2281
AATGTTGACA TGTCAATGTT AAATGAATCA AGGCTACCAT ACCGCTTGAT TTGAGTTTTA


 


2341
ACACTATCCA ATCCATATTC TTCAAGTAAT GCTTTTAATT TACCAGTAAC TTTTTCACCT


 


2401
AAATGCCCAA AATACTGAAG CCAGCTCTTA AGTAATTCTC GGTACTGGTC AAATTTTTCT


 


2461
TTTTCAGTAA GCTGTTTCTC AACTTCTTCT TGGTCCTCAG AACCGTTAAC TACTTTCCAG


 


2521
AGGTCAGGGT CAGTAGTACC CAACTTATAA CCAAGTACCC GATCATATGC TCTGGCCTTT


 


2581
TCCTTTGGAG TCAAATCAGG ATTAAGGTCA ATATTATCTA GCTTCATACG TTGGTCAGCA


 


2641
TAACGACCTT TTTCAAAGTC ATCGGCATTC TTGGCTTCAG GTTGCCAAGT GAACTCATAG


 


2701
CCAACAACAG GACGTCCTCT ACCAGCTCCT CGAATAGTTT TTACGGTTAA GCCACGAATA


 


2761
ACAGGGGTTA GCTCTTCTTT AATTGGACGT AACACTCTTT TTCTTACGTT ACCCTGGTCT


 


2821
TTCTTGTAGC TCTTGGGTAG ATCTAGTTGC TTAAATAATT CGTCACTGGT TAACTTCAAC


 


2881
CGCCCTACTG TTCGATATTG TTTAATTAAC ACTCTTTTTC TTACGTTACC CTGGTCTTTC


 


2941
TTGTAGCTCT TGGGTAGATC TAGTTGCTTA AATAATTCGT CACTGGTTAA CTTCAACCGC


 


3001
CCTACTGTTC GATATTGTTT AATCAAGCGA AACATGTTCT TAGCATAAGC AGATTGAAGA


 


3061
CTATTAAACT GAGCTAGTTG AAATCTTGTC CAATGACTTA AATCATTGAA TAGCTTCTGA


 


3121
AATAGAGGGT TAACTTGAAC AGTCAGAATC TGTTTACTCC TCTGAATCTG AAATACATTC


 


3181
CAACAAGCAA ATTGAGTAAT GGTGTCTCCA TCATCGGTGT AGGCATTAAT CTCTAATAGC


 


3241
TTTTTATTAG TTTTCATCAA ATCTTTTACA AAATTAGATG TACTCCGTTC ATACTTACTA


 


3301
AGACTTTGTA GTTGCGTAAA TGAAAACTTT AATTTCTGAG TACCCTTTTG ATACGACTGC


 


3361
TGAACTAGTG TAATAAATAG ATTTAGCTCA TTAGAGTTCA GTCGGCCAAG TGGAATAGTA


 


3421
TTAAGCCGAT TACTATACTT CACAATTTCA TTACTCAATT TTCTCACCTC AAATATATTA


 


3481
TAACATTTTA TCGTACACAA TCAAGAGTAC GTACTTATAG CACACGTACA GTATTAGCGC


 


3541
GTATATTAAG ACAAAAATTT AGACAAAATG TACCTTATAA CGGACAAAAT GTACCTTATA


 


3601
AACCGGACAA AATGTACCTT ATAACGGACA AAATGTACCT TATAAAAAAC GAAACTCTTA


 


3661
CTCTCCCAAG GAGTTTCAAA CCCCTAAAGA GTATTTAAAG AATATATATA AAGATATTTA


 


3721
AAGAGGCACC ATACGGAAAA TCTCCTTTCT GATTTCAATT TCAGAGAGGA GATTATCACT


 


3781
AAATTTCAAA ATTCAATTTT TCGCCAAAAA CTTTTTTAGT AGTTTTCGGT AACTAGCAAA


 


3841
ATCAACTTCG TTGATTATTT TGACTATTTC GAAATTCATC AAAAACTGAA TGCCCGTCTT


 


3901
TATAGTGAAT TTGTTTAGTG AAGTTCACTA ACTGCTTCAG CATATTCAGT TGACCACGTA


 


3961
ACCATTGTTC ACTATCAGCA GGACTAATTT GTTCTGATAA TTGCTCTTTT TGGTGGTTAT


 


4021
AAGCCTTATA AGCCTGATAT TTAGCATAAC CTTGTGGGTC ATCTGATTTA GCCACTCTAG


 


4081
CCACTTTAGG TACTTTTCTA AGCCCCTTAG CTTGAGCTTG AATATCTTCT AAAACCGCCC


 


4141
CTTTTTCGAT GAGACGCTTA TTTCGTTGCT TTCTCTTTTC AGTATTGATG CGAGCCATTT


 


4201
GACGCTCTCG ACGAGCCTTA AACACTGCTT CCTTTTCTTG AAGGCGTTCA AGTTCTGCTT


 


4261
TCATTTTTTC AATATCAGCC ATAACAAATC CTCCTAATGT GAATAATGTT TTCTGAAATT


 


4321
CAAATGTTAT CTAGATTATA TGATGTTACA ATTGATTTAA CCACTACGTT TGGTCTCCCT


 


4381
GTTAAGGGCG CACTTATACA CATTCAAAGA ATGTAATTAC GCCTGACGGC GACGTGCGCT


 


4441
CTCCGAGGGG ACTTACCCAG TCGGGGCGAC TTCATCGAGA CCATGTCATG GAGATTCAGT


 


4501
TTCACCGAAA CGGACCACTA ACCCCGTGGA GGGGAGAAGT TTCACTTCTA GGGACCATTC


 


4561
CATGGAGGGT CGCTTTTTAC AAAAGCAACG CAAAACAAAA GACTAGCTCC CAAGCTTACG


 


4621
CAAGGTCGTT AGTCCCTAAG GAGGCAGCAC AGA.






A putative L. reuteri plasmid origin of replication identified based on BLAST analysis could be:









(SEQ ID NO: 8)










1
TTAGTCATCT TCTGTCTCCT GTTTTAGAGA ATTTTCAATC ATGTCTAATA ATGTTGACAT



 


61
GTCAATGTTA AATGAATCAA GGCTACCATA CCGCTTGATT TGAGTTTTAA CACTATCCAA


 


121
TCCATATTCT TCAAGTAATG CTTTTAATTT ACCAGTAACT TTTTCACCTA AATGCCCAAA


 


181
ATACTGAAGC CAGCTCTTAA GTAATTCTCG GTACTGGTCA AATTTTTCTT TTTCAGTAAG


 


241
CTGTTTCTCA ACTTCTTCTT GGTCCTCAGA ACCGTTAACT ACTTTCCAGA GGTCAGGGTC


 


301
AGTAGTACCC AACTTATAAC CAAGTACCCG ATCATATGCT CTGGCCTTTT CCTTTGGAGT


 


361
CAAATCAGGA TTAAGGTCAA TATTATCTAG CTTCATACGT TGGTCAGCAT AACGACCTTT


 


421
TTCAAAGTCA TCGGCATTCT TGGCTTCAGG TTGCCAAGTG AACTCATAGC CAACAACAGG


 


481
ACGTCCTCTA CCAGCTCCTC GAATAGTTTT TACGGTTAAG CCACGAATAA CAGGGGTTAG


 


541
CTCTTCTTTA ATTGGACGTA ACACTCTTTT TCTTACGTTA CCCTGGTCTT TCTTGTAGCT


 


601
CTTGGGTAGA TCTAGTTGCT TAAATAATTC GTCACTGGTT AACTTCAACC GCCCTACTGT


 


661
TCGATATTGT TTAATTAACA CTCTTTTTCT TACGTTACCC TGGTCTTTCT TGTAGCTCTT


 


721
GGGTAGATCT AGTTGCTTAA ATAATTCGTC ACTGGTTAAC TTCAACCGCC CTACTGTTCG


 


781
ATATTGTTTA ATCAAGCGAA ACATGTTCTT AGCATAAGCA GATTGAAGAC TATTAAACTG


 


841
AGCTAGTTGA AATCTTGTCC AATGACTTAA ATCATTGAAT AGCTTCTGAA ATAGAGGGTT


 


901
AACTTGAACA GTCAGAATCT GTTTACTCCT CTGAATCTGA AATACATTCC AACAAGCAAA


 


961
TTGAGTAATG GTGTCTCCAT CATCGGTGTA GGCATTAATC TCTAATAGCT TTTTATTAGT


 


1021
TTTCAT.






Analysis of the strain 3632 sequence reveals two agglutinin receptor precursors (ARP) that are uniquely found in L. reuteri strain 3632 compared to L. reuteri 2091, strain 170331 and L. reuteri ATCC 23272 (human). These agglutinin receptor precursors likely result in the increased level of auto-aggregation observed with this strain, and these receptors would contribute to the probiotic potential of a DFM genetically modified to express them. The amino acid sequence of ARP-1 is:







(SEQ ID NO: 9)


MNEYNAEMAKLNQGANAPVITTNSVNQALSLKPENNATVDIEALNPRIT


 


FKRVEEGTKYAGYQIFDKNNAYVNNIDGEFLRVTYTNLKNSTYKGSKIS


 


KIVVTYSDSTPTGNRITQSGLNAVTEGANDNFLVVFEDPVRGDMHSTTV


 


TATYQYYDANGNLIDFSGTNNAWLSVGSLNFDQGNDYQGGKNEGNPTSG


 


ISEGVKLISGAQIKQLAGSSISVHDDGWAYAGFNNYSGTGMNNGINTDN


 


GGSGWDMDGSPNAYYGAIVFQLTGSSVSLRQGLVSWGGADIASQYNNQF


 


LNNAWFTAGTTLPETQIKQPIRKTSETHYHYNPSVIRL.






The amino acid sequence of ARP-2 is:








(SEQ ID NO: 10)


MAQKLMSANSTDKNFKMYKSKKSWVFAYSTTLALAAVAGITLSTTNVHA


 


DTTNGGDNQVNATAVTQNTTSNTVDQIAANTAQTDNTSTSINIRSLMDD


 


LASGDDTSSSQNGQEQSQNYASSNQNSQTQQENGTTGQSTASQNGTTSD


 


QTNSDQSDKNYYVISTRDLDKNGNVNYLTQKNYTSIKGQEVADGTVVTW


 


PLSVSALPANRAQDLKSHVISETLDPHLEYLHYRAYLTNTDGTVTDVTN


 


HVNLNRSGQTLIFTDDNYLLSIYNNNRYRVQNLPVIKLVTKANGNGYII


 


PNAFKSSYVFNDGSHDVSFTTTSNNVQIKTFNPGNSKDVEIGGNVQGDP


 


SGTINGQVVADGSVVTWPMSVGDLPANRAQDVLSHIETDTLYNGLNYEG


 


YHAYLPQADGSFQDVSSHINVQQNGQDLTFIADDYLIGLYNQDKSTAFK


 


MPIIDLITSVHGTSIIAPNKFNSQLAFKDGNGQTVINNTSNQVQISTYH


 


PTNTKDVELGGNVQGDTPNSINDKVVANGAIVTWPMASSELPANRVQDL


 


QSRVISETLDSHLQYQGYKAWLQNADGKYTDVTSHVKLTQDGQNLTFAD


 


DEYLLNLYNSNKGTAYKLPIIDLVTKVNGAGITAPNSYTTKYVYSDGDG


 


NTTINVTSNTVKISTFNPTTNKDVELGDNIHGDTESSIAGKLVSEGTIV


 


TWPLSTSDLPANRAQDVVSHTAVDALEPTLQYISYTAWLPDSNGQLQDV


 


TSHVKMTRDGQKLTFTDDDYLIGLYNQNKDIALKMPIIDLVTKATGNTK


 


LLPNSFDSQFVYNDVDGNTIINVSSNKPTVETFDPTVHKDVELGGNNVQ


 


GDTPNSIDGKIVAQGTVVTWPMSTSDLPANRTQDVVSHSTSETLNQNLQ


 


YVGYHAYMPDANGKLQDVTSHVQLQQNGQNLVFTDDSYLINLYNQDKSI


 


AFKMPIIDLMTKAISDSATIPNTFESQYVFNDGNGNTTFKSTSNTVQII


 


TYKPKTTKDVELGDNIHGDTNASIAGQMITDGTVVTWPMSTSDLPANRT


 


QDLQQHVVTDNLNDNLIFQGYTAWLPTANGLVDVTNHIELTRDGQNLTF


 


TDDAYLLNLYNQNKDTAYKLPIIDLVTKANGNTKLIPNNFDSMFVYNDG


 


DQQTTVNVTSNTVNISTYDPTATKDVELGDDIEGDTADTINNLMVQIGT


 


KMTYPLTVSDLPANRADEITAHQSVDTLSDYLEYQGYKAYLPDADGKLQ


 


DITEHVNLKREGQKLSFNDDDYLINLYNNSKATKQALPVIDLVAKVTGS


 


NDGKKVHIIPNHFDSTITTKDGKINTTSNTVVINSNDPEAVKDVELGDN


 


VVGDTPNSVTGTTVADGTIVTWPMSVGSLGANRAQNVIKHTETENLDSG


 


LTYLSFKAYLPDADGKMQDITEHINIQQDGQKLVFTDDDYLISLYNKDK


 


SQRFALPVIDLVTRVNGDNKIIPNTFVSQFTFNDGKGNTITSVTSNQVN


 


VSTFKSNPEKHVTLGTDIEGDDAENADGTVVAQGSEVTWPLSDKSPLPA


 


NRSQDVKSHTLVDKLDDNLQYNSYKAYLKGTDGKLQDVTDHIKLTRDGQ


 


NLTFIDDDYLLDLYNKDKSTAFNLPIIDLVTTVVGNDKLIPNKFDSNFV


 


FSDGNKDTSMKTTSNEVSISTYTPVTNKDAELGDNVVGDTSDSIANETV


 


PDGTIVTWPLSVSSLPANRSQDVFKHVIEDILDGNLTYNSFKAYLKDAA


 


GNLQEVTDHVKLAQEGQHLTFTDDDYLINLYNSSKNKEQSLPIIDLVTT


 


VHGDSKLIPNEFDNVFVFKDGKGQTTVKTTSNKVTIKTASLPTPTKEET


 


DDQGNNINGNEVKAGEHVNYTLNWDLSNDKDVKATPEMIKKGFFFIDPI


 


DSRALSVDDLSKAKVVDQNGNKVDGISFHLYNSLSEVPEFIQEQVKANN


 


LQDKITGPFVVAQADDLQAFFDKYVKTGAKLKVTIPTIVKSGFTGEFSN


 


TAYQFGFGKATPTNTVTNYVKPMHKPASPETPAAIAPQVISATAQPMTS


 


DAPVTPSEKTAKLPQTGNADEGALLGLAAVSLVGSLGLAALGLKQNRND


 


D.






A person of skill in the art would recognize that, because of the redundancy of the genetic code, multiple nucleic acid sequences could encode the above peptides.

L. reuteri strain 3632 contains eight bacteriophage-encoding loci, but only three of these loci appear to encode productive prophages. Bacteriophage loci 1 has a sequence of:








(SEQ ID NO: 11)










1
CTATTCAAAT TTGAAATTTT GAACAACTTT ATCAAGGTCT TCTTGTTCAA TCCCTAGATA



 


61
GTGGCGCGTA TTTCGTTCAG AGGAGTGATT AAATAACTGC GAAATGATTT CAACGTTAAC


 


121
ACCTTTCTTA TAGAGTTGTC GACCAAAAGT TTTTCTAAAA GAGTGTGTTC CAATTTTAGC


 


181
AACGATTTTA TTATTTTCCG TTTTTCTAGC CATTCGTTGC AACATTTCGT AAAAGCCATG


 


241
TACTGAAAAA TGACCTTCTT GTTTCCCAGG GAAAAGATAA TCATTCTCGT CTTTATAATT


 


301
CAAGTCATTA AGATAATCAA TAATTTCGGC AAGACAATTA TTCCAAAAGA GCGTTTTAGC


 


361
CTTACCTGTT TTTTGCTCAA TAACACGGGT TTTAGTTTTA TTTAGAACAT GACCAACCCT


 


421
TAACGTTACA ACGTCTGATG CTCGCAATCC GTTATTAAGG GCAATTGCAA TTAGTAATAG


 


481
GTTTCGGTTA ACCAATTCAG GACGCGAATG CAGCTTAATT GTCTCACGTA ACTGATCATA


 


541
TTCTTCGAGA GTCAAGAATT TAACATTTTC TACCCATATT TTCCGTTTAC CCTTGATTTG


 


601
ACCATTAGTT TTATTTTTGG GAATATAAAC TCGTTTTTTA GTTGACATAA TATTTCTTCC


 


661
TTTGTTAATT TGCGAGGTAA TCAACTAAAG CAGGGACGAT TTCCTCTCCT TCTTTCCGCC


 


721
CAGTTTCATA AAGTGCTCGA ATTTTCTTAA TATTTCCTTC AATCCGCTTA ATTTTAATTG


 


781
GTTTAGAGGG AGCAATATTA AAGATCTTTC CTTCACAAGA CAAACGATTA ATTTCAGCAG


 


841
TCTGTTGGTT GTAGAGAAGG GGACGATCAA TCCCCGCTTT AGCAAATTCG GGATAATCTT


 


901
TATATACCAT GTCGTACAAC TTCTTTACAG CAGAACTAGA TGGCTTCTTT CGGTAGTTAA


 


961
TATCCCTGGT CCGTACAACA ACAATCTTGT CATAGCCTTG TCGTTGTGCA ATATCAAATG


 


1021
GAATTGAATC TGTGATCCCA CCATCAAGGC AAGGCCCTTG AGAAGTTTCC TGTGGATCTG


 


1081
AAAGAAATGG CATTGATGAG GAAGCTTTGA GGGCATTGGT TAGCTCCTCA CCAACAGGAT


 


1141
CCGTAAATAA AACTGTTTTT CCTGTTTTCA CAGAAGTAGC AACTGCTGTG AAATGAGATG


 


1201
CTGACCGTCG ATATGCAGCT TCATTAAAAT TCTGCCAAGA ATATCCGTGA TCTTCAAAAA


 


1261
GATAATCAAG ATTAATTATT TGCTTTTTAA ATACGCGAGC CATCGAGATA TATTGGCGAT


 


1321
CGTGCCGATG ATCAATATTA ATATTGGCAG CGCGTCCGTA TTGCTTAGAG ACGAAATTAA


 


1381
CTCCACAAAG TGAACCAGCA GAAACGCCAA TTACACTTCG AAATTCAATA TGGTGAGCGA


 


1441
GAAATGTATC AACAATTCCG GCAGTATATT GTCCACGGAA TGCGCCACCT TCTAAAACTA


 


1501
ATGCTGCATT ATATAACATA AAATGAAATA ACTCCTTTTT GTCCGCCAAA AATAGCAAAA


 


1561
ATGACCTAAT TTTTAGTGAG ACACTTTTTC AATAAAATTA TACAACAGTC TCACACGAAA


 


1621
ATATAGGATT TTAAAAGCTT GTTATATCAA CGTTTATTCT CGGTTACCGT AAGACTCTTT


 


1681
TATCAAAAGA GTCTTACTGA TTTTGAAATA ACGCAAAAGT AGGGGCAAAA ACGGATTTTG


 


1741
CCTCTCAGAC GAACTGAAAT TCATTTTGAT GAGTAGTAGG TCGTTACAAT TCGTCTCAGA


 


1801
AGAAAAACGC ATATTGATAT CAAAAAATGA GTAAATAGAC AGTAAATATT TAAATATGAA


 


1861
CTTTTTATGA AGAAACTATT GATAATTTAA GATAATCATA ATAAAATAGT GCACAGTTAA


 


1921
TTGTTAAACG TTTAGCAAAA AAGGTAAAGA AACGGGGTTA TTTCTATGCT AGAACGCAAG


 


1981
GAACATAAGA AAATGTATAA AAGTGGTAAA AATTGGGCAG TTGTTACACT TTCTACGGCT


 


2041
GCACTGGTGT TTGGTGTAAC AACTGTAAAC GCATCCGCAG ACACGAATAC TGAAAACAAT


 


2101
GATTCTTCTA CTGTACAGGT TACAACAGGG GATAATGATA TTGCTGTTAA AAGTGTGACA


 


2161
CTTGGTAGTG GTCAGGTTAG TGCAGCTAGT GATGCGACTA AAAATTCTGC TAATGCAAAT


 


2221
AGTGCTTCTT CTGCCGCTAA TACACAAAAT TCTAACAGTC AAGTAGCAAG TTCTGCTGCA


 


2281
ACAGCTTCAT CTACAAGTTC AGCATCTTCA TCAACTAACA CAGATAGTAA AGCAGCTACA


 


2341
GAAAATACTA ATGTAGCCAA AGAGGATGAT ACACAAAAAG CTGCACCCGC TAACGAATCC


 


2401
TCTGAAGCCA AAAAAGAACC AGCTGTAAAC ACTAATGATT CTTCAGCTGC CAAAAACGAT


 


2461
GATCAACAAT CCAGTAAAAA GAATACTGCC GCTAAGTTAA ACAAGGATGC TGAAAACGTT


 


2521
GTAAAAAAGG CGTGGATTGA TCCTAATAGT TTAACTGATG ACCAGATTAA AGCATTACAT


 


2581
TAAATAAGAT GAACTTCTCG AAAGCTGCAA AGTCTGGTAC ACAAATGACT TATAATGACC


 


2641
TTAAAAAGAT TGGTGAGGCT TTAGTTGATC AAGATCCTAA ATACGCTATT CCTTATTTTA


 


2701
ATGCAAGTCA AATAAAGAAT ATGCCCGCTG CATATACTAG GGATGCTGAA ACAGGTGAAT


 


2761
ATGCTGATTT AGATATTTGG GATTCATGGC CAGTTCAAGA TCCAGTGACT GGTTATGTTT


 


2821
CTAATTGGAA TGGCTATCAA CTTGTGATCG CTATGATGGG ACGGCCTCAT CACGAAGATA


 


2881
ATCATATTTA TTTACTTTAT AATAAGTATG GTGATAATGA TTTTTCACAC TGGCGAAATG


 


2941
CAGGATCTAT ATTTGGATAT AATGAATCTC CACTAACACA AGAATGGTCC GGGTCTGCTA


 


3001
TTGTTAATTC TGACAACTCT ATTCAACTCT TTTATACTAT CAATGATACA AATAATGCTA


 


3061
TTAACCATCA AAAATTAGCA AGTGCTACTA TGTACTTAAC AGCCGACAAT GATGGTGTCC


 


3121
ATATTAATAA TGTAGAGAAT AATCATGTGG TATTTGCAGG TGATGGTTAT CATTACCAAA


 


3181
CTTATGATCA ATGGAAAGCT GCAAATAGTT TTGCTGACAA CTACACTTTG CGGGATGGAC


 


3241
ATGTTGTACA AATGCCAAAT GGTGATCGGT ATTTAGTATT TGAAGGAAAT ACTGGAACTG


 


3301
AAAATTATCA AGGTGAAGAT CAATTATATA ATTGGTCAAA TTATGGTGGT AACGATCGCT


 


3361
TTAATATTGA AAGCTTGTTT CATCTTTTAA GTAGTGATGT TGATTATAAA AAAGCTATCT


 


3421
TTGCTAACGG GGCACTTGGA ATTATTAAAC TAACGAATGA TGAAAAGAAC CCGCAAGTAG


 


3481
AAGAAGTATA TACTCCATTA GTTACATCAA ATATGGTTAG TGATGAACTT GAACGTCCTA


 


3541
ATGTTGTTAA ACTTGGAGAT AAGTATTATC TATTCTCGGC TACACGTTTA AGCAGAGGAA


 


3601
CTAATATCGA TACTCTTAAT AAAGCTAATA AAGTTGTCGG AGATAATGTT GCAATGATTG


 


3661
GTTACGTAGC CGATAGTCTT ACAGGTCCAT ATAAACCATT AAATGGTTCA GGGGTTGTAG


 


3721
TGACAGCTTC TGTTCCTGCC AATTGGCGTA CCGCTACTTA TTCTTACTAT GCTGTTCCAG


 


3781
TAGAAGGAAA AGAAAATCAA TTACTCATTA CTTCGTATAT GACAAATAGA GGCGAAGTTG


 


3841
CTGGAAAAGG TATGAATTCA ACATGGGCAC CAAGTTTTAT TGTGCAAATT AATCCTGATG


 


3901
ATACAACTAT GGTATTAGCT AAAGTAACTA ACCAAGGGGA TTGGATTTGG GACGAATCAA


 


3961
GTAACAATAA TAATATGTTA GGAAACATTC AAACAGCTGC TTTACCTGGG GAATTTGGTA


 


4021
AACCAATTGA TTGGGATTTA ATTGGTGGTT ATGGATTAAA ACCGCATGAT CCTGCTACAC


 


4081
CAAATGATCC TGAAACGCCA ACTACACCAG AAACCCCTGA GACACCTAAT ACTCCCGAAA


 


4141
CACCAAAGAC TCCTGAAAAT CCTGGGACAC CTCAAACTCC CGATACACCT AATACTCCGG


 


4201
AAGTTCCTTT AACTCCAGAA ACGCCTAAGC AACCTGAAAC CCAAACTAAT AATCGTTTAC


 


4261
CACAAACTGG AAATAACGCC AATAAAGCCA TGATTGGCCT AGGTATGGGA ACATTGCTTA


 


4321
GTATGTTTGG TCTTGCAGGA ATTAACAAAC GTCGAGTTAA CTAAATACTT TAAAATAAAA


 


4381
CCGCTAAGCC TTAAATTCAG CTTAACGGTT TTTTATTTTG AAAGTTTTTA TTATCGAAAA


 


4441
AAACAAATCC TCGTTAATCC TTTAATGCAA TTGTTGTAAA ACCTTGCGAC AGTAATAACA


 


4501
GTGGATTTGC CCATCTTTGT CAGCTAACTT CCGTGCATGC ATTGCAGAAA AAGTATAGTG


 


4561
CTCATGACAA AATGGACAAA CATATTGCTT TTTCCCAAAT AGTGAGGTAA TAAAGCCCAA


 


4621
AATTTTCTCC TCCATAAAAA AATTATACAC CCCTTAACAT TATAGCGAAG ATTATCTTTA


 


4681
TAGAAAAGGC TTTTATTCAT TTGTTTTTTT ATATGAAGTT TCACTAATTT CTTTAAGAGC


 


4741
ATCACGCTGC TGTTTCTGGT CAACGTGAGT ATATAAATCG GTAGCAGATG TTCCTTTTTG


 


4801
TCCCAACTGT TGGGCCACTA ATACTTGATC TTTCGTTACC TCATACATTT CAGAAGCAAG


 


4861
GGTGTGCCGT AATTTATGGG GAGTAAGAGG ATGGCCAAAT GCGGTTGAAT ACTTTTTGAC


 


4921
CATTTTTTCA ATTGCATTAG CTGTCATCCG TCGAGTTTGC TTGTGATAAA CGGTTAGGAA


 


4981
GAAAGCAGTA TCCTTTTTTA AAGCGTGGTA TCGTTGTGCA CGAATCGCTT GGTAAGTTTG


 


5041
GATATAAGTA ATAGCCCAAG GAGCAATTGG TACCGAATCC CTCTGGCCAC CTTTTCTTGT


 


5101
TACATCTAGC AGCGATTGTT TTAAGTTTAA ATCACCAAGG TTTACATTTG CCGCCTCGGA


 


5161
TACCCGGACG CCTGTTCCTA AAATAAGAGC AATAATTGCA ATATCTCGTT CCTTATTGAC


 


5221
CTTATAAGAG GGGAGAGCTC TTTTATCACA TTTATTAGGG TATTCTTTTT CAATGAAAGT


 


5281
AATAAAATCA AACTTCATTT GTCCCCGGTA CATGTGTGAA GCCAGCGTAT GAGCCCGATA


 


5341
ATTTAATGTT TTTGTATCAT TAAGGGAGTC AATTTTCAGC ATTACATTAC GGTCAAAATA


 


5401
AGATTCACCA TTATTGTTAT CTGCGGTAAC TGTTAAAAAT TTATACAATG ACCTTAATGC


 


5461
ATTAATAGAT CGATTGATTG AAGTAGGAGA ATTTAACCGT CCTTGTGCAT TGGTAGTATG


 


5521
CTGCAAATAA TCGATATACA ACATTACATC GCTACGCCGT AGATTCGCCA GTGTATCAAT


 


5581
TGGTAAGTCC TTATTAGAAT TAACAGAGAC GAGTCCACTT GACCGTAACC AATCAAAGAA


 


5641
TCGACGGATT TCAGTTAAGT ACTGATAGGT TGTTGTGACT GCATGGTTGG TCCCTAAATA


 


5701
ATACTCGTTA ACATAATCAG GAAGGTTCTG AAGCTCCTCT TGAATTAGCT TTAAATATTT


 


5761
ATCTGCTTCC ATTTTTATAT CCTCCTTGAT ACTAAAAAGA GTCGCTTGAA AGAAAACTTT


 


5821
CAACGACTCT CTAAACCTAT CGGGAAAACA GGATTCGAAC CTGCGACCCC CTGGTCCCAA


 


5881
ACCAGGTGCT CTACCAAGCT GAGCTATTTC CCGTTAATAA CGAACAAATA TTATTCTACC


 


5941
AATTCTCAAT TAATTAGTAA AGGAATATTT TAATTTAAAG TGATTAATAG TTAATGATTA


 


6001
AATGACATAA TAGTAAAATT CCTATTAATT GCAAAAAAAT CATGATTAAC CGAAATCCTT


 


6061
TTCTAGCATC TAATGAAGAT GATAAATAAT TTTATTTTCG ACTAGTTATA AAAGATACCT


 


6121
TTCCGAAAAT AATTTGACAT TCAAAATACT TTTGAATATA ATTTGATTAT CGAATATTTT


 


6181
GATACTCGAA ATATTTTCCG AAGGCAGGTG AATCTTTTTG GCAATAATGA ATGCGCAGGA


 


6241
GATTATGGAA TTAATTCCTA ATCGATATCC GATTTGCTAT ATCGATTATG TAGATAAATT


 


6301
AGTTCCTGAA GAAAAGATTA CCGCAACAAA AAATGTCACA ATTAATGAAT CATTTTTTCG


 


6361
CGGTCATTTT CCTAATAATC CTGTTATGCC TGGAGTTTTA ATTATCGAAA CATTGGCCCA


 


6421
AGCTGCTTCA ATTTTGATAT TAAAATCACC GCATTTTTAT AAGAAAACAG CTTATCTTGG


 


6481
CGCAATTCAT AAAGCAAGGT TTCGACAAAT GGTCCGTCCT GGTGATGTAT TAAAACTAAA


 


6541
CGTTGTTATG AAAAAAGTTC GATCATCAAT GGGGATTGTA GAAACACAAG CGCTTGTGAA


 


6601
CGGCAAATTA GCTTGTAGTG CGGAGCTTGT CTTTATCGTT GCTGAACGAG AAGAAAAGAT


 


6661
TTAGCACGGT GAATCATTAT CATTTATAAT ATATTTTGAT AATCAAATTA TTCGGGCTAT


 


6721
AGAATAAAAT TGAAGGGAAG AAATATATTA TGAGTAAAGA TAATGATTAC GAAAAAATTA


 


6781
ATAAAGGATT AATCAAAGTT TATTCCGGAA TTCTATGGAT TGAAGAAAAT GAATTGCGGA


 


6841
AAAGTACATT CAATGATTTA ACAATTAAAG AGATGCATGC AATTGATGCA ATTACGATGT


 


6901
ATAACCATCA AACAATTTCT CAAGTAGCAG AAAAGCTTCA TCTAACTCCA GGAACAATGA


 


6961
CTTCAATGGC TGATCGTTTA ATTCGAAAAG GATATGTGGA AAGAATTCGT GATAAAGATG


 


7021
ATCGCCGCAT TGTTCGGTTA TGTTTAACCA AAAGAGGCCG GGTACTATAT CGTGCGCACC


 


7081
GGGCCTTTCA TAACATGATG GTTGAACGTT TTCTTAAAGG AATGGATGAT GAAGAAATGA


 


7141
AGGTTGTTAA AAAAGCCTTG CAAAACTTAG AAGATTTTGT GGATGAGCAT GCTTAGAGAT


 


7201
AAGGATTGAC AAACATTGCA AAATTTAAGA ATTACTAGTA CTGCGAGTTA TCATCCACCC


 


7261
CTTAAAATTA CTAACCAGCA ATTATCAACT ATTATGAATA CTTCAGATGA GTGGATTAAG


 


7321
ACGCGGACTG GAATTCATCA ACGTTATATC AGCAACACTG AAAATACTTC AGACTTAGCT


 


7381
GTCAATGTTG GTACCCAGTT ATTGACTAAT GCCAATTTAA AGGCAACTGA ACTTGATTTA


 


7441
ATCATTATTG CAACGATGTC TCCTGATGCC TATACTCCTT CAACTGCTGC TATTGTTCAA


 


7501
GGAAGATTAG GTGCGAAAAA TGCAATTGCG TTTGATATCT CAGCAGCTTG TACTGGTTTT


 


7561
ATCTATGCCA TTAATACAGC TGAATTAATG TTGAAAAGCT CTCATTGGAA AAATGCAATG


 


7621
GTAATTGGTG CAGAAGTATT ATCAAAACTG ATTGATTGGC AAGATCGAAG TACTGCCGTA


 


7681
TTGTTCGGCG ATGGGCTGGC GGAGTGTTAC TTCAAAAGAC AACTACAACA ACCCCTTTAA


 


7741
TTCTCGGCCG TGATCTCCAT ACATTTGGTG ACTTAGGAGA TAAAATTATT GCCGGAAAAA


 


7801
CAACGCCTAA AACTGGCTTC CCTAAACAAC TAACATCCCT TTCACCATTT ACGATGGCCG


 


7861
GCCGTGACGT ATACCGCTTT GCCACTCATG AAGTACCACG ATCAATCACT TCTGCCGTTC


 


7921
AACAAGCTAA TTTGAAATTA GACGATATCG ATTATTTTTT ATTACATCAA GCAAATGAAC


 


7981
GGATAATTAC CCAAATTGCA AAGAGACTGG CGCAACCAAT TACAAAGTTT CCAATGAATA


 


8041
TTAGTGAATA TGGGAATACA GCTGCCGCTA GTGAACCAAT TTTATTGACT CAAGCTATTG


 


8101
CTCATGAATT GATTAAACCA GGTAACATTA TTGCAATGAG TGGCTTTGGT GGCGGGTTAA


 


8161
GTACAGGAAC AATAATTTTA AACTATTAAT AGAGAAAGAA GAATGGATAT GACTAAAGAG


 


8221
GAAATTTTTA ATACTGTAAA AACTATTACT GTTGATGAAT TAGATGTTGA CGAAAATCGT


 


8281
GTAACAATGG ATGCACGAAT CAAAGATGAT CTTGATGCTG ATAGTCTTGA TGTTTTTGAA


 


8341
ATTATGAATG AGCTTGAAGA TAAGTTTGAG ATTGAATTAG ATGCCGATGA AGGAATCGAG


 


8401
ACTATTAGTG ACGTTGTTGA TTTCGTAAAG AAACAGTTGG ATGAAAAATA ATGTACTACG


 


8461
GAATATTATT TAGCGGTCAA GGTGCACAGC GATCGGGAAT GGGAGTCGAG CTTATGGCCG


 


8521
ACTCCCTTTT TTCAAGGATT GTTAGTCAGG CAAGCGCTGT TTGTGAACTT GATTTGCTCA


 


8581
AAATTATGAA AAACGAGCAT AAGGAGTTGA ATAAAACAGC ATATGTTCAA CCAGCAATTG


 


8641
TAACAGTTAG TTATGGAATC TACCGAATGT TAAAACGGGA TTTACCTCAA CTACCAATTA


 


8701
AAGGGATGAT TGGCCTATCT TTAGGCGAGT ACGCTGCTTT AATTGCTAGT AATGCACTTT


 


8761
CATTTGAAGA AGGAATTAAA TTAGTTGCGG ACCGTGCTCA CTTTATGCAA CAAGACGCGG


 


8821
ATAGAGAAAT AAGTACATTA GCAGCTGTCC TTGATCCTCA ACTTCAAGAG ATAAAAGAAC


 


8881
TAATCACCGC TCAACAAGAA AATGGTCAGC GAGTTTATAT TGCTAACTAT AATTCACCAC


 


8941
GACAAATCGT AGTGGGCGGG GCATTAAACG CTTTAAAGGC TACCCTTAAA AAGATTGAAG


 


9001
AGGACAAGCT TGCTAAAAGA ACGATCCTGC TCAAAGTTAA TGGTGCATTT CATACTCCCT


 


9061
TCTTTAACGG TGCACGTCAA CAGATGCATA ACCGGTTACA AGCAGTCGAC TTCCATGAGC


 


9121
CACAGATTGA AGTTATTAGC AATACTACTA ACAGCTTGTT TCATTGTGAG GATCTTCCAG


 


9181
GAATTCTTGA AAAACAATTA GCTGTTCCAA CACACTTTGG AGCTAATGTT AAGGAATTAG


 


9241
TCAAGCACGC GAAAATTGAC ACAATATTGG AAATTGGTCC TGGAAAGACG TTATCTCGCT


 


9301
TCGCTCACCA AGTTGACCAG CACTTAAATA CCCAACACAT TGAAAATCTT GCTGATTATG


 


9361
AAAAATTTAT AAAGGAGCAA AAAGATGGAA CTGACAGATA AAGTCGTTTT TATAAGCGGA


 


9421
AGCACACGCG GGATTGGGGC GGCTACTGCA TTAGAGTTTG CTAAGGCTGG TAGTCGGCTA


 


9481
ATCCTCAATG GGCGGCAGGA TAACTTACCA AAAACGTTTA AAGAAAAGCT AGATCTACTA


 


9541
GGGGCGGATT ATCACTATCT TAAGGGCGAT ATTGCAAATG AAGAATCAGT TAGTGAATTA


 


9601
GCAGCAGCGG CTTGGCAAAT ATACGAGAAG ATCGACATTC TTATCAATAA CGCGGGAATC


 


9661
ACGAATGATA AGTTAATGAT GGGAATGAAA GCGAGCGATT TTGACCAGGT CATCAATGTT


 


9721
AATTTACGCG GAACATTTAT GTTAACGCAA CCTATTTTTA AGAAGATGCT CAAAAAAAGA


 


9781
GTCGGTTGCA TTATCAACCT TGCTAGTATT GTGGGTCTCC ATGGTAATAC GGGACAAGCT


 


9841
AATTATGCGG CAAGCAAGGC AGGTATCATC GGCCTTACTA AATCTATTGC CCAAGAAGGA


 


9901
GCACGCCGTG GAATTCGTTG CAATGCGATT GCTCCCGGAA TGATTACTAG TGATATGACT


 


9961
GAAAAATTAT CTGAGCGAGT AAAAGAACAA ATTCTCAGTC GCATCCCCCT CAACCGCTTA


 


10021
GGACAGTCAG AAGAAGTTGC TAAGACCGCA AAATTTTTAG CAGAAAACGA TTATTTGACT


 


10081
GGTCAAACCA TTGTAGTTGA CGGTGGCATG ACAATTTAGG AGGAACTAAA TGACAAGAGT


 


10141
TGTAATAACA GGAATGGGTG CTGTTGCTCC TAATGGTAAC GGTATTCAAG AATTTATAAG


 


10201
TAATAGTTTT GCAGGCAAAG TTGGAATTAA AGCGATCAAG AAATTTGATG CCAAGTCGAC


 


10261
AGGAATTACC GTGGCAGGTG AAATTGACGA TTTTGACCCT AATGATGTCA TTGGAAGGAA


 


10321
AGCCGCGCGC CGAATGGATC TTTATTCTCA ATACGCCTTA CAAAGTGCGA TTGAAGCAAT


 


10381
GGAAATGGCG GAGATTAACG AAACAAATAC CAAGCCAGTC GACATGGGTG TTATCTATGG


 


10441
ATCTGGAATT GGCGGTTTGA CAACTATTCA AGAGCAAATT ATCAAAATGC ATGATAAGGG


 


10501
TCCTAGACGG GTATCACCAA TGTTTGTTCC AATGTCAATT GCTAACATGG CAGCCGGTAA


 


10561
TATTTCCATT CACTTTAATG CGCAAAATAT TTGTACATCG ATTGTGACTG CTTGTGCCAC


 


10621
TGGAACTAAT GCAATTGGTG AAGCCTTTCG TCAAGTTAAA GCAGGTCGCG CTAAAGTAAT


 


10681
GATCGCTGGT GGATCAGAGG CTTCGGTGAA TGAGATCGGG ATTGCTGGTT TTGCGGCATT


 


10741
AACAGCATTA TCACAAGCAA CTGATCCGCT TAAAGCTTCT TTGCCATTTG ATAAGGCACG


 


10801
TCAAGGATTT GTTTTGGGTG AAGGCGGTGC AACACTTGTT TTAGAGGATC TTGAGCATGC


 


10861
GCAAAAACGC AGTGCTAACA TTCTTGGTGA GATTGTCGGT TATGGTGCTA CCTCGGACGC


 


10921
TTACCATATT ACATCCCCTG ATCCAACTGG TGCAGGGGCG GCAAGAGCAA TGGAACTGGC


 


10981
AATTAAAGAA GCTGGAATTA GTCCTAGCGA AATTTCCTAT ATTAATGCCC ACGGAACCGC


 


11041
TACTCATGCT AATGATGAAG GCGAATCCAA AGCAATCAAT CAGGTATTTG GTTCCGATAG


 


11101
TAATGTTCGC GTTAGTTCAA CGAAGGGAAT GACCGGCCAT TTGCTTGGGG CTGCGGGCGC


 


11161
AATTGAGGCA GTCTTAACAG TAGCCGCTTT ACAAAAGGGA CAATTACCGT TGAATATAGG


 


11221
TTGCTTTAAT CAAGATCCAA AATGCTCGGT TAACCTTGTG ACGGCAGAAA ATAGGAACGC


 


11281
CTCAACCGCC CGTTACGCAA TAAGCAATTC TTTTGGTTTC GGTGGTCATA ATGCTGTTTT


 


11341
AGCCTTTAAG AAATGGGAGT GATCTATCTT GGAATTTAAA GAAATTCAAA CATTAATGCA


 


11401
AAATTTTGAA GATTCTGATA TTCGTGAATT AGAAATAAAT CAAGATTCCT TTCAGCTCTA


 


11461
TTTAAGCAAA AATAAGCAAA CCCACAAGCA TGAAAATCTT ATAACAACCG AAAAAACAGA


 


11521
GCAAACGACT TCAGCTAAGA AAAAAGCAAA CGAACAACTA ACTTTACCTT CGCAAAATAT


 


11581
AACTGCGCCC CTAGTCGGAA CAGTCTATCT CCAACCAACC CCCGATGCAG ATCCCTATGT


 


11641
TAAAAGTGGC GACCACGTAA AAAAGGGGGA TGTTGTTTGT GTGATTGAAG CAATGAAAAT


 


11701
GATGACAGAG ATAAAAAGCC CTTTTAACGG AACCATTACT TCAATTTGTG TAAGCAATGA


 


11761
AGAATTAGTT GAAGTAGAAC AACCGCTTTT CTCAGTTCAG GAGGATAAAG ACAATGCCTA


 


11821
ATAAAACTTT AGATATAACT GAAATTCAAA AAATCCTTCC GCATCGTTAC CCGATGTTGC


 


11881
TAATTGACCA AGTTGATGAA TTAATCCCTG GTAAGAAGGC CATTACACAC CGTAATGTCA


 


11941
CGATTAATGA AGAGGTTTTT AATGGTCATT TCCCCGAAAA TCCAGTTTTA CCAGGAGCAT


 


12001
TGATTGTTGA ATCATTGGCG CAAACAGGTG CCGTCGCTCT CTTATCTCAA GAAGAGTTCC


 


12061
AAGGGAAAAC AGCCTATTTT GGTGGAATTC GGTCAGCAGA ATTTCGTAAG GTAGTCCGTC


 


12121
CGGGCGATAC GTTAAGGTTA GAAGTCAGAC TAGAAAAAGT TCACAAAAAT ATTGGAATTG


 


12181
GTAAAGGCAT TGCAACGGTC GATGGCAAAA AAGCCTGTAC AGCCGAATTA ACTTTTATGA


 


12241
TTGGGTAGGT GGTTTAATGT TTTCCAAAGT ACTAGTGGCT AATCGTGGTG AAATTGCTGT


 


12301
CAGGATAATA CGGTCATTAC GGGAGCTAGG AATTAAGACA GTAGCTATCT ATTCAACTGC


 


12361
AGACCGCGAA AGTCTTCATG TTCAACTAGC GGATGAAGCT GTATGCGTTG GAACTGCCCG


 


12421
GGCCCAAGAT TCATATTTGA ATGCGAAAAA CATTTTAGAA GCTGCTCTTG GTACAGGTGC


 


12481
CCAGGCAATC CATCCTGGCT TTGGCTTTCT ATCAGAAAAT GCGGAATTTG CGACAATGTG


 


12541
TGAAGAATGC GGAATTACGT TCATCGGTCC CCAAGCCTCA GTGATTGACT TAATGGGAAA


 


12601
TAAGGAGCAT GCACGGGAGC AAATGAAAAA ATCAGGGGTG CCTGTGATTC CTGGAAGCGA


 


12661
TGACTATATT ACCAATGTTA ATGACGCTGT TGAGGTCGCA AACAAGATTG GGTATCCAAT


 


12721
TTTGTTAAAA GCAGCTGCTG GTGGTGGCGG TAAAGGGATC CGACGAATTA ACGATCATAA


 


12781
CCAGATGCGG CAAATATTTA GCGAGGCCCA AAACGAAGCC CGACTTTCGT TTAATGATGA


 


12841
CCGAATGTAC CTTGAAAAGA TTATGGAGAA TGTTAAACAC ATTGAGGTCC AAGTATTTCG


 


12901
TGATAATTTT GGCAATGCCG TTTTCTTTCC TGAACGTGAC TGCTCGATTC AACGGAATAA


 


12961
ACAAAAATTG ATTGAAGAAA GTCCTTGTGT CCTAGTAAAT GAACAAGAGC GAAAAACGCT


 


13021
AGGACAAATT GCAATGCGAG CCATTAATGC GATTAACTAC CATAATACGG GGACAATAGA


 


13081
ATTTCTAATG GACAAGGACC ATCACTTTTA CTTTATGGAA ATGAACACTC GTATCCAGGT


 


13141
TGAACATACA GTGACGGAGA TGGTAACTGG GATCGACTTA GTGAAGGCAC AGGTTATTGT


 


13201
CGCTGCGAAT GAACCACTTC CCTTTACCCA ACAGGATATT CAGGTTCATG GACATGCAAT


 


13261
TGAGTGTCGG ATAAATGCTG AAAATCCTAA ACAAAACTTT ATGCCAGTGA CTGGGACGAT


 


13321
TAATTACTTA TATCTTCCAG TCGGTAATTT GGGGATGCGC ATTGACACTG CTATTTATCC


 


13381
TGGCAGTAAG ATCACTCCTT ATTATGATTC AATGATCGCT AAGGTAATTG CCCTTGGTCA


 


13441
AGATCGCCAA GAAGCTATTG AAAAAATCAA ACGACTTTTA AATGAAATGG TAATTACGGG


 


13501
CGTAACGACC AATCAAAATT TCCATTTAGC AATCCTAAAC AATCCTAAAT TTTTGGCAGG


 


13561
AACAGCTTCA ACAACGTTTC TTGAAGACTT CTTCTTGCCA CAATGGAAAA AGGAGCTGAC


 


13621
AGCGTGAAAT TATATGATCA AAATAATACT TTAAGTGAAC GGCACATCAA AGCAGATAAA


 


13681
AATGCTGATG AAAGGGTCCC AGATCAAATG TGGTTAAGGT GTCCACATTG TCATCAATTA


 


13741
CTATTCGCCA AGCAGTTAAC ACAATATGCT GTTTGTCCCA ATTGTGACTA TGGATTACGA


 


13801
ATACCTGCCC GCCATCGACT CTCGTGGTTA GTAGATTCAT TTAAAGAATT CGATAAGGAT


 


13861
CTCCAGACAA AGAATCCGTT ACATTTTCCT GGATACCAAG AAAAAATCAG CAAACTTCAA


 


13921
AGACAAACTA AGCTGAATGA TTCAGTCCTA ACTGGTGAAG CTTCAATTAA TGATCAGCTA


 


13981
TTTTCACTAG GTATTATGGA TCCAACATTT ATTATGGGAT CCCTCGGAAC TGTTACCGGT


 


14041
GAAAAGATAA CGCGCTTATT TGAATATGCA ACTATCCATC GTCAAGCAGT AGTACTATTC


 


14101
ACTGCTTCAG GCGGGGCACG GATGCAAGAA GGAATTATGT CGCTAATGCA GATGGCGAAG


 


14161
ATTTCACAAG CAATAAATGA GCATGCTGCT GCCGGCCTTT TATACATTGT GATCTTAACC


 


14221
GATCCAACAA CTGGTGGAGT AACAGCTAGT TTCGCAATGG ACGGAGATAT TATTCTCGCT


 


14281
GAGCCCCATG CACTTGTTGG CTTTGCAGGC CGTCGAGTTA TTGAACAGAC GATTCATCAG


 


14341
CAAATTCCTG TTGATCTCCA ATCAGCTGAA AACATCCTGC ATCATGGATT TATTGATCGA


 


14401
ATTGTAAAAC GTCAAGATGA AAAAAAGCTG CTTGAATGGC TATTAAAAAC AGGGAGCGTT


 


14461
GCTAATGAAT GAACAATTAT CAGCAAGCGA GATTGTTAAA CGTGCTCGCA ATGACAATAA


 


14521
AATTACGGGG ATGGAGATTA TTCAAAATGT TTTCCCAGAT TTTGTTGAGT TGCACGGCGA


 


14581
CCGGGCAGGC GGGGATGATC CTGCAATCGT TGGTGGAATC GCTACTTTCC ATCAGCAAGC


 


14641
AGTTACCGTC ATTACCACTG ATCGAGGAAA AACAACAGAA GAAAAAATCA TAAAGCATTT


 


14701
TGGCTCACCA ATGCCTAGTG GTTATCGCAA GGCACTCCGC TTAATTAAGC AAGCAGTTAA


 


14761
ATTTAAGCGA CCTGTATTCT GTTTTGTTAA TACCGCAGGA GCATTTCCTA GCAAGGAAGC


 


14821
CGAAGAAAAT GGGCAAGGAA GTGCGATTGC CCAAAACATT TTACAAATAA GTCAGCTTGC


 


14881
CATTCCAATT ATCACGATTA TTTATGGTGA AGGAGGTAGT GGGGGAGCCT TAGCATTGGC


 


14941
ATGTGGAGAT GAAGTATGGA TGTTAGAAAA CAGTACTTAT TCTATTTTAT CTCCTGAAGG


 


15001
GTTTGCCTCC ATCATGTGGA AAGATAGTAC GAAAGCAGAT AAAGCGGCAG AATTAATGCA


 


15061
AATGGTGCCG CAAGCTTTAT TAAAACAAGG GATTATCGAA GGAATTATTC CAGAAAGCGA


 


15121
AGAGCATCGT AAAACTTGCA AAAATATCGA GCAGGTTTTA CTAAAGCGAT TAAACAAGCT


 


15181
GCAAGAATTA CCGCCAAACC AACTTCTAGC AAACAGAAAA AAACGTTATC GAAAGTTTTA


 


15241
AGGAGGATAA TATGGGAAAT ATATTAACAG GAAAAAAGAT CGTTGTTATG GGAGTAGCTA


 


15301
ATAAGCGTTC TATTGCATGG GGATGTGCAC AAATGATGGC TGAACAAGGT GCCCAAGTTA


 


15361
TCTATACTTA TCAGAATTCC AGAATGAAAA AAAGCTTACA ACGGTTAGTA GATGATGAAG


 


15421
ATCAATTAAT TGAATGTGAT GTAGCAGATG ATGAAAGTAT TGACCAAGCC TTTACGATTA


 


15481
TCAAGGAACG TTTTACAAAA GTAGATGGAA TTGTACACGC AATTGCTTTT GCAAAAAGAG


 


15541
AAGAATTAGC TGGTTCAATC CTTGGTGCTA GTCGCAAAGG TTATGCAATT GCGCAAGATA


 


15601
TTTCGTCTTA CTCCCTTATT GCTGTCGCTA AGGTTGCTAA TGAGTTAAAT CTATTAAATA


 


15661
ATCCTGCAAG TATTGTTACC TTAACTTATT TTGGCTCAGA ACGTGCTATC CCTAACTATA


 


15721
ATGTAATGGG AATTGCTAAA GCTGCCCTTG AAGCTAGTGT TCGCTATTTA GCACGGGATT


 


15781
TAGGACAAAA ACGAATCCGT GTTAACGCAA TCTCTGCTGG CGCGGTTAAG ACATTAGCAG


 


15841
TTACAGGTAT TAAAGGTCAT GATGAACTTT TAAAGATGTC CCAAGCAAGA ACTGTTGACG


 


15901
GAGAAGATGT GACTATTAGC GAAATTGGGA ATGTGTGTGC ATTCTTAATG AGCGATTTAT


 


15961
CAACTGGAGT TACTGGCGAC ACCATTTATG TTGATAAGGG GGTACATTTG ATTTAAAAAT


 


16021
ATAAATTTTA AAGACTGAGA AATGAGTTTT TTTCAGTCTT TTTGCTGTCT TTTAGAAGGA


 


16081
AAACATAACA ATAGCAAGAA CTTTTTTTAA TTTTTTCTAA AAAGTTCTTG CTATTTTATA


 


16141
GCTCAGTTGG TATTATTAAT AACGTTGTGA AAGACGAGTT AATAATTTTG AGAAAAATTG


 


16201
TTTAATATAT TTAAAAATGG CCCGTTAGTC AAGTGGTTAA GACACCAGCC TTTCACGCTG


 


16261
GTATCGTGGG TTCAAATCCC GCACGGGTCA CTTTTGCGGA AGTAGTTCAG TGGTAGAACA


 


16321
TCACCTTGCC ATGGTGGGGG TCGCGGGTTC GAATCCCGTC TTCCGCTTAG CCAGTTCTAT


 


16381
TATGCCGGGG TGGCGGAATT GGCAGACGCA CAGGACTTAA AATCCTGCGG TTAGTGATAA


 


16441
CCGTACCGGT TCGATCCCGG TCCTCGGCAC TATTTCGGGA AATAGCTCAG CTTGGTAGAG


 


16501
CACCTGGTTT GGGACCAGGG GGTCGCAGGT TCGAATCCTG TTTTCCCGAT CTGGCTAAAC


 


16561
AGCTAAGAGG TAATCTTCTT AGCTGTTTTT TTGGATATCT GTCAACTGGT ATGGTTGACT


 


16621
AAAAATTTTT TTAGTGCGCC CGGCATGGGT ATTAGCTAGG TGGTGAAAGT CCGCTATGGG


 


16681
CCGTAGTAGT CGGAACCATG AGCTGAGGAC AAGGGTGTCC ACCGTGAGGT GGAATCTGAA


 


16741
GGAAGTCTAA GGCAAAGTAC TGCATCGATG AACAAGAAGT AGCTATAAGG CTGAAATTAA


 


16801
CTGGATAAGG CTGCTAGACA AGTTGAAGTC CAATACTACT CGAAGTTGGT CTCAGTAAAG


 


16861
CTAACGATGA CATGGTACGA AAGCTAATAT TCTTACCCGG GGAGATCTGG CCTACACGTT


 


16921
TCCGACAAGC AGTACTAACG CAGCCATTAA TTAGTTTTTT GAAATAAAAT AAACTTTTTT


 


16981
GAAAAAAGTA TTGCTTTTTA TAGGATAACT TGATAATATA ATATTCGTCG CTGATATGCG


 


17041
GGTATAGTTC AGTGGTAAAA CCACAGCCTT CCAAGCTGTT GTCGCGAGTT CGATTCTCGT


 


17101
TACCCGCTTT TAATTAATTT AATATGGCCC GTTAGTCAAG TGGTTAAGAC ACCAGCCTTT


 


17161
CACGCTGGTA TCGTGGGTTC AAATCCCGCA CGGGTCATTT TTGGAGGATT AGCTCAGTTG


 


17221
GGAGAGCATC TGCCTTACAA GCAGGGGGTC ACAGGTTCGA GCCCTGTATC CTCCATTGAA


 


17281
CAATTTTAAT AGTTGTTCAA CATTTTAATA TAATTGGGCT ATAGCCAAGC GGTAAGGCAA


 


17341
CGGTTTTTGG TACCGTCATG CGCTGGTTCG AATCCAGCTA GCCCAATTAG TCTAAAACAG


 


17401
TGAGTAAAAA TTACTCGCTG TTTTTTATTA GAAAAAATAG GCAATCTAAT AACGTAGGCT


 


17461
AAAAGCGAAC ATCATTGGAT TGCCCAATAA TTAGCATAAT TTATTCACGT CGTAACATAA


 


17521
TATGCCAGAC AAAGAAGCCG ACTCCAATAA TTAACGCCAA CCATGCCCAT AGTTGCAATG


 


17581
GAAGATAGAT AGTACACGTT AACAAGCCTA AAACCCAACT GATTAGCATC ATCGATAATC


 


17641
CAGCAATATC GCCACCTAAA CCATCAATAC CACAAGTAGC GATATAGACG ATTCCTGATG


 


17701
CCAGATATAA TAGCGAAACA ATTAATTCGG CTGTTCCGGC ATAAATAGGA ATTGCAGGAT


 


17761
TAGGATTCCC AAAACCTCCT CAAGTCTTAT TCTACCCGGA TGATTAGAAT TTATAAAGAA


 


17821
TTACAGTTTT TCTATTTACA AATAGAGCAG GGGGAACAAA AAATCAAACC AGAAATTATC


 


17881
TTCTGATTTG ATTGGATCTT ATTTAGTTGT ATCTTTTACT TTTTTCAAGG CTTCTAAAAA


 


17941
TACTTCATAA GGTTGTGCAC CAGTGATTGA ATACTTATTA TTAATCACAA ATAGCGGTGC


 


18001
ACTTGGCATT CCAATCATAA ATGCGCGCCG TTCATTTTTT CGAACTTCAT CTTCATACTG


 


18061
ATTAGATTCA AGAACTTTTT TAACATCAGC AACAGGTAGT CCGATTTCAT TCATTGCAAC


 


18121
AGTTAGAGCT TCATAGTCCG CGATTGATTC ATTATCATTA AAATAAAGTT GATAAAGACG


 


18181
TTTAACAGCC TTATTAAGTA ATGCTTGGTC ATTTAAACTC TCAATGTATT TTATTAAGCG


 


18241
ATGAGCAGCT AATGTATTAA CAGGAATCGC ATTTTCCATC TTAATTGGTA AATCAATATC


 


18301
GGCAGCAAGT TGATCAATTT TATGTATTTG TGTAACAGCT TCTTGCTTAG TTAGTTGATG


 


18361
AGTTTTTGCA TAATATTCTG TCATTGATAA ATCAGTTGTT GTTGGCAACG TTGGATCTAG


 


18421
TTGAAATGAC ATAAATTTTA ATGGTGTCTG ATCGGCAATC TTTAATTCTT TAAGCGCCCG


 


18481
TTGTAATTGT GTAATTCCCA TATAGCAATA TGGGCACGCA ATATCAGCCC AGTATTGAAT


 


18541
TTCCATAGAT GATCGCTCCT TATAAATATA TATTAATTTA ATCTAATCAT AAAATAAGCG


 


18601
AAGTTGCTAG TAAATACGTA AAGAGGGAAC GATTTATTTT AAAATTAACA TGTCATTAAA


 


18661
ATTATAGTTT TAATGCGACT GAATTTAAAA TTCCCCCCAG AATATCAATT TTTAGCTTTC


 


18721
TCAAAAGAAT TTATTTAATG CTTATTTTTA ACTTTAATAA AACTTTTTCT AAACAAAACT


 


18781
AATACTTTTG ATTTATGTTT TAAAAGATTT ATGTAATACT ATTGATGAAG TCTATGTCAA


 


18841
AAGTATTTTA AAAGGAGTTT TTATCATGAA ACATACGCTT AAAGTTGATC AAGTACGTGA


 


18901
CGGTTTATGG CTAGATTCAG ATATTACGTA TACGCAAGTT CCTGGATGGC TTGGTAATAC


 


18961
AACGCGAGAT TTGAAGCTTT CAGTCATTCG GCATTTTCAA ACTAATGATG ATACACGTTA


 


19021
CCCAGTAATT TTTTGGTTTG CTGGTGGCGG CTGGATGGAT ACTGATCACA ATGTTCATCT


 


19081
GCCAAATTTG GTTGATTTTG CTCGGCATGG TTATATTGTT GTTGGTGTCG AATATCGTGA


 


19141
TAGCAATAAA GTTCAGTTTC CTGGGCAATT GGAAGATGCT AAGGCTGCTA TTCGTTATAT


 


19201
AAGAGCTAAT GCCAAGCGCT TTCAAGCTGA TCCTAATCGG TTTATTGCGA TGGGAGAATC


 


19261
AGCTGGTGGT CATATGGCAA GTATGTTAGG TGTTACTAAT GGTCTTAACC AATTCGACAA


 


19321
AGGTGCTAAT TTAGATTACT CCAGTGATGT TCAAGTAGCG GTTCCTTTTT ATGGTGTAGT


 


19381
TGATCCCTTA ACCGCTAAAA CAGGAAGTGC ATCAAACGAT TTTGATTTTG TTTACCGTAA


 


19441
CTTGCTTGGC GCTGAACCTG AAGCTGCAAA TCCCCTCACG TATGTAAATT CTACTTCTAC


 


19501
GCCCTTTCTT ATCTTTCATG GTACGGAGGA TGTCGTTGTT CCAATCAAAG ATAGTGAAAA


 


19561
GCTTTATGAT GTATTAGTTG AAAACAACGT TCCTACTGAA TTATACGAAA TTGAAGGTGC


 


19621
AAGTCACATG GATGTAAAAT TCCTTCAACC ACAGGTATTT AAAATTGTGA TGGACTTTTT


 


19681
AGATAAGTAT TTAACCCGGC CATAGATCAT TTCTTGCCTT TACTACTAAA AAGCAAACTA


 


19741
ATAACAATCC AACTTTCACA TTATTGACAA AATCAAAAAG CACCATGAAA TTACTTTTTC


 


19801
ACGGTGCAAC TATTGTTAAT ATTTATCTAC TTGGCTTCCA TCAATGGAGA AATAGACAAC


 


19861
TTGATTTGCA AACCTAATCC AGCAGCATAT TTATTTAATG TAGGCAGCGT AGGGATTGAA


 


19921
TCTAAATTCT CGATTTTAGC CAATTGAGGT TGTGTCATAC CGATTTTTTT TGCAAATTCT


 


19981
GTTTGCGAAA TGCCCTGTTT AATTCTTTGT ACTTCTAAAA AAGACAAGGT ATCAACAATC


 


20041
GATAATTCTT CTTTTGTTGC AGCCGTCTGA TGCTTATCTA TATCTTCCCA TTTTCTCATT


 


20101
ATTTACCCTT CCTTTCGTAC CAATCGTCAA GTAGCGACAA GGCTCTTTCT ATCTGACGAG


 


20161
GATCTGTTTC ATCCTTTTTC TTAGCATAAT GATTTAATAA TACAAAATTA TTTTTCTTCC


 


20221
ATACTCCATA AAATACTCTT TCTGGCATAG GTCGTAACTC CCATAGTTGA TGCCTATAAC


 


20281
CCTTTAATTT TTTTGCTTGT GGTGTATGCA ACACAGGACC AAGAGCTTGT AACATCTTAA


 


20341
TTTGATGGCG CATTTTTAAA TAAATTGCTT TATCTTGCTT TTGTTTACTT TGCGAAATTT


 


20401
TCTCGAAATA ATCACCAATT TCACTATTTC CATTTTTATC TTCGTAAAAA ACTACTTCAT


 


20461
ACATGCACTA TTTCCTTTAT TATAGCTTAT TTATTACTAA TTATAGCAAG ATTGCTATAG


 


20521
CTTTACTACT ATTAAAATAT ATTCCATAAT CAAAAATTTT TCTATTGAAA GCACCCGCAC


 


20581
ACTCAATTAA ATCAAATAAG TTTACGGCAA TGATTATTTT TGGTGCACAA AAATTGCCGA


 


20641
TGTTCCCCAG CTTTACAAAC TTTCACCATT TCATACCATT TTAACTAAAC AAAAGCTCCG


 


20701
GTAAAATGCC GTTATATCAG CATTTTATCG GAGCTTCTTT TTTATACAAT CTTAAAAAAA


 


20761
TGCGTCCCCC GAGCATAGAA GATTGTTGAT ATATCAACGT TTTGAAGGAG TTAGTGTGCC


 


20821
AAACGTGTGC GAACTTAACT AAAATAAAAA AGAGCGCCTA TCAGGGCGCC CCACGTTATA


 


20881
CCGGTACGGA TCAAGTAACG AACTGAATCC AATTTGCGTG CCGATACTTC TCATCTGATA


 


20941
TTTAAATTAT AACATAATTA TTTTATGATA CAATAATTAA TGCGTGTCGG GAGTAACGAC


 


21001
CCAAACTATC ACGCAAAAGG GGTGAATTCC CCGTGACATT TATACTTCTC ATTCTGATCG


 


21061
TGCCGAACGG GCTCGTCAAG CAACTGATGC ATTATGCAAA GGTTGTAATC AAAAAGGCTC


 


21121
TCAAACATCT AATTGATGAA TTACTTGGTT AATTCTGATA GGAAATGTGC GAGATGCTAT


 


21181
TTCAGCGGTA GTGTTTGCTA TCGCTGTTTT TTATTATACT TTAAGAATAT AGTCATGGTA


 


21241
TTTTTTTACC TTAGTAACTA TTCAGTTTAA TGTTACTCTA TTAAATTTTG AATTCCTTTA


 


21301
CATCCTTAAC CTCTATCATA TTACAATCTT TATCTTCAAC GTAGACGAAT TCTGACATAT


 


21361
CATCAATATC ATTGAAATCT TTATTAGCTG CGAGTTCTGC AGCATCCTTA AAAAAATTAG


 


21421
GCCTTTTTTC TTCTATTTTG TCCATCGGTA CTAAACATTT TAATATGTAA GTTTTGAAAT


 


21481
GCTCTTGTTC CCATCTTGTT TGAAGAGGCC CTTTTAATGG CCTACTTCTA ATATTATCTA


 


21541
ATTCATATAT GGCACTATCA AGGTACCTTA AAATTTCAGG ACACTTATAA ATGCAAGAAT


 


21601
AATCAAAATG ATATGGTATA AAAAAGCCAC TAATTCCGTT ATCATTTTCC AAACGTGCTG


 


21661
CAATATCATT CCAACTTCCA CCAGATTCAA TCAGGTATGA GTGCCCATTA TATGAAATAA


 


21721
AAGGATTATT ATTTTTACAG TAGAAACGAA TATTATTTTT TTCTAAAAAT GATCCTAACA


 


21781
AGGTACTAGA ATCTGTTATA ACTTTGCTAA GTGGTTTTAA TCCATCATAT AAATAGTTCC


 


21841
TTTGCTGTGA AGAACAAGTT GTAAAGTGGT AAAAATATGC ATTTTCAATT TCATCTTTTT


 


21901
TATATAGTTC TAATGATCTT TCAGCCATTT CATGAGTCGA TAAATCAAAT ATCTTGTATT


 


21961
TTTTCTTTTC AGACACCATG GTTTTCTCCT TTTTATGATA TATCTAATTT AGATTACTAT


 


22021
ATGACAAAAC ACCCTAGCAA CCATTAATCG GCTACTAGGG TGTTTATTAC TATAATTTTT


 


22081
CACTTAAATC TGAATTGTTT GACCAATGAA AATCATATTA GGATTTGCGA TGCCATTCTT


 


22141
TTGAGCTAAT GCTTGCCAGG TCTTGCCAAA CTTAGCCGCA ATTCCGCTTA GTGTGTCCCC


 


22201
TGCTTGGACA GTATAAGCAT TGGATTGTCC GTTGCCGGCT AAGTAGAGTT TTTGCCCAAC


 


22261
GTAGATCACA TTCGGGTTAG TAATGTGATT ACGGCTTACG AGGTCTGAGA CAGTGGTGCC


 


22321
GAATTTGGTG GCAATTCCGG ATAACGTATC GCCTGATTGA ACAAAGTAAG TATTTTCGTT


 


22381
TGATACTTTT CCGGTGACTT TGAGGACTTG GCCGACATTG ATCTGGTTTG GATTACCAAT


 


22441
ACCATTGATT GCTGCTAGGT TCTGATAAGT AGTCCCGTAT TTTTCTGCAA TTCCACTCAA


 


22501
TGTATCGCCC GGCTGAACAA TGTAGGTTCC AGTAGCTGGG TGACCAATAT GTTGAACCGG


 


22561
CTGTGGTGCT GGAATAACAG CTTGCGGAAC ATTGCCTGAT ACTAAACCAG TCGTAAAGGC


 


22621
ACCGTCAAAG TCATAACTAG TATCAATTCC CATGATGCTA TGATCAGAAT ATTGCCAAGC


 


22681
ACTTGCATTA TCAATCCCCA GTTCAGTCAC ACCATAGCCA GCAATCCAAA TCTTCCGGGA


 


22741
ATCAAAGCCA TGTGAATTAA GAATACCGCC AGTGAAGAAG GACTTCATGG AGTAAATCCC


 


22801
AGTATTCTTG TAACCAAGAG CTTCTACTTC TTGAAGAAAG GCTAAGGATA CTGATTGATA


 


22861
ATCTGCAGTT GAATGAACTT CCGCATCATC AATCATCAAA GTATCGTCAT ACATACCAAA


 


22921
TTGCTTAGCG ATTTTAACGA AGAATCGAGC TTCATTTTGT GCATCAGTCA CTGATGTATA


 


22981
ACGAGCAAAG TGGTAACAGG AAACGCGCAA GCCAACCGCT AAGGCATTAC GAATTTGAGC


 


23041
GGCCGCACGT GGATTAACAT AAGCCGAACC ATCTTCGGAC CCTTCCGTTA ACTTAACAAC


 


23101
GACCCCTAAT GCACCCTGAG CTTTAGCAGC TTGGAAAAAG GCAACAGTAT CTGGTTGATA


 


23161
ACTTGAAACA TCGATGAATT GATTACGCAT ATTATTCTTC CCCTTTCATT GGATCGATTG


 


23221
CTGGTGCTTG GCCGGCTGGA ATAGTAGCGG GTTTAGTTGG TTGTGATGTC GGATCAACGG


 


23281
TTGGTGTCAA TGCAGACTTT TCATAAGCTG ATTGCACAAG CGTTTCAATA GCGTTAAGGT


 


23341
CAATATTCTT AATACCCTGC TTTTGAAGTG CTTGTTGAAC AATACCTGTT GCTTGATGAA


 


23401
ATTTCTCATG ACCAGCCATG TCTTTGCCAA CTAAGGAAGC AACGGCATTA TCTGCAAGTC


 


23461
GTTCAGCACA AGACCAAGCA GCACGAGAAG TTTCTGTTTT GGCATGAGTG ACTTTAGCGT


 


23521
GAATAAATGT CTGGCTTTGC TTTAAGGCAA AATAAAAGAC CGTTGAGATA ACGGCCGTCA


 


23581
TGATATAGGT TGGAATTGCA TTAATGATTG TGCTCATGTT GTTTCTCTTC TTTCTCTACA


 


23641
TAATCTTTGC TGAGCTTGTA AATATGATTC TTTACCCAAG ATGGTAATGG TAGTCCCATT


 


23701
TGGCCCCAGT TTTCGATAAT CGAAACTAGG TAAAATAGGA AAAAGAAAAA GACAAGCGAG


 


23761
TCGCCTGCCT GTCCTAATCC GCATAATTCT AACATTGGAT AAACGGTTAA AGTCAAAATA


 


23821
ATAACTGCAG CATGTTTCAA AAGTCCACCC GTTCCTTTAC TAGAAGTTGT TCGGTGCGTA


 


23881
ATGATACTTT TGAAAAAACC TGTCACAATA TCAATCACAA TCGCCCAAAC TAACCATTCA


 


23941
ATGAGCACGT TGTCAATCAT GCTGGCAAAA TATTGAATGT ATAGAACGTG TTGTGGCGGT


 


24001
TGTGTCAGTA GTAAGTGCAT TTATAGTGGT CACCTCCTTA TAAGGGTAAA ATAAAAACGC


 


24061
CCTGTAGAGG ACGTTTAAAA TTATTTAATT TTTATTAAAA GATTCCCAAT TTCCATCTTC


 


24121
ATTATTATTG ATTTTAATTA TTAAGTCGTT AAGTTTATTA TCAATATCAT TAGCTAGCTT


 


24181
ATACGCACTT GAATCACCTC GTACTCTAGT AATTTCATAT ATTTTTCCTT GACGAAGTTC


 


24241
TTCAGTCAAA GATTTAATAT TATCTATTTC GTTAGAATAT TTCTGTTTGA GATTCACTTT


 


24301
ATCCCCATCT CCTTACCGCT AAAATAAAAG TGTTATATTA CGCTGTTACT AAATATGGAA


 


24361
TATTATTATC AGATAACCAT TGAACTGCAT TATTATTAAT TTTTACAACA TCATCTAAAA


 


24421
TTTTATCTAA ATTAAATGAA TAACCAAATA TAAAAAATTC ATGCTTATGA GGATCGTATA


 


24481
CTCTTAAAGT TTCATCTTTT GTAGGAGTGA ACACTTTGTC TCCAACTTTT ATAGTTTGAT


 


24541
ACAAATTACC AATTACTTGT TTTTCAACCA TTATGTGCTG ACGTCGGTTA AATTTAATAA


 


24601
GTCCTTCATC AATATGTTCT AAACTATTCC TGAGGCCTCT TAAATTTTCA TTTTCGACCC


 


24661
TATTAATATA GAAATCATCA GTTACATTCA AAATTTCCCT TACCTTTTTT CGATCATTAC


 


24721
TATTTTTAGA GTTAGAACCC CATAGAAGCT TAGCTATATC TGACGTATAC AATGCAATAT


 


24781
TTTGTAAATT ATACCAAATC ATATTATCTT TATAATATTT TTCATGTTTG GTATTATATA


 


24841
TATTTTTAAT AGCAATAATA GTAAAATTAC ATAACAGATA AGTTTCACTA ATTGCATACT


 


24901
CCTCGTCTTT ACTATTCATA GTATAATGTT GCATAATATT TACCATTCCC CTTATTAATT


 


24961
GTTTAGTAAT ATTATCGTAT CAGTAATTGA AATAATTATG ATATTATTAT GAAAAATTGA


 


25021
AGAAAAACCA AAGGCTAATA ATTAATTTAC AAAAGTATGG AATCAAAATT TTAATAAAAT


 


25081
GCCGCCCATA ATAAAAGCCC CGCTCGTTTG AGTGAGGCTT ATTTATGTAT TGTGTATTTC


 


25141
TAAGGCGACT ATAAGTAATT ACCGTTAAGC TAAGAATTCA TTTAATTATT AGCCATTTTC


 


25201
TTTAGCTTTT CTTTATCTTC AATCTGATAC AAGCGAATCT TAGTAATTAA TCCCTGATAC


 


25261
GCTTTTACTT CATTGGGAAA ATTATTAGCA GGAGTAAATT TATCATCTTC TTGGGACAAT


 


25321
TCAGAAACAT AGAATACATT TCGATAATAA CCTATATAAA GTTTATGGTT CAGTTTCCTA


 


25381
TATAATTTAT TTTTTTCAAT AATTTTTGCT TCTTTAGGAG TATAACCATC TTGTTTTCGG


 


25441
TCAAAATAGG CTTCCCATTT AATGCACTGT TCCTGTGAAT TAATCCGATT TGTTTTTAAA


 


25501
CCAAATGCTG CTAATGGATA CCCACCTAAT TTGGGATTGT TACACAATTC ATTATCATAT


 


25561
TCGTAAGCAG TATTAACAAT ATCTTCTAAT GATTTTGAAA TATTCAAGAT AACCACCTCT


 


25621
ATTTTATAGA GTATCACTAA TTTAGACTAC TAAACAATAA CTTAAATAAT TTTATTGTCA


 


25681
TGATTTATGC TCCTTGAATT TCGTAATTCT GACCGGTTAC CGTCTTATAT TCATCTTTAG


 


25741
TGATCGCTCC GCATCGTACA TAAACTTTGT AGAATTCTAA ATCATGGTTA CCCCAGTCAT


 


25801
TCCAAAACAT TTGAAGCATT TGTAATTGTG TCATCATTAT GCCGTTACCT CCTTAGATTT


 


25861
TTCGACTGCC TGTTGCTGAT TAACAGTCAT AAACATTTGT TGTAATTGCT TGATTTGAGT


 


25921
TGCTTGTTGA GCATTAGTAG CCTGCAATTT TGCATTTGAT TGGTTCTGTT GCATAATCAT


 


25981
TGCTTGCAAA ATAGTGATTT GTTGATTGGT ATTCCCGTTT GAATGCGAAA TAATCGTCAC


 


26041
AGGAATTTGA CTATCATTCA TAATAAAGCT CTCCTTAGTA TATTTTTGTT TGATAAAACA


 


26101
CTGTTACAGT TACCCCTGTA TTTTGTATAG CTGAGTTATC AATGTTTCTA ACGATTAATC


 


26161
CATCGCCGTT TCCAGCTTCT GAAACACTCA ATTGCCAGCG GTCGCCTTCT GCAACATAGT


 


26221
TAAGTATTTT CCCAGTAACT CCAAGTTCTT TATTGATAGC ATCCATTGGA AGTGTCTTGA


 


26281
CCTCTTGCGA ATTAAGTTGA TCAATAGCTA TAAATTTAGT AATCGTCTTT AAATTAGAAT


 


26341
TGTAATCTGT TTGATGATCT TGCAAACTCA TTCCTAGCGA TGTATTGCCT AAACTAATGA


 


26401
GCGAAGTTGG CGTTTTATCA GCATAAATTC TCCCAGCTTG AAAATGGGCT GAGTTTTGAT


 


26461
CGTAAAATTT AACTCCAGTA TTAAAAGCCT TTCCACCTAA CTTAAAGTGA TCCACATCTA


 


26521
GATATGTTCC AGTCGTACTA GGCCCAATAT ATAGCCCATT CATGCTTTCT CTTTCAGCTC


 


26581
CTAGCATAAT CTCGTTTGCT GTAAACTCAG CGTTAGTTTG CAGAGTGAGC AACTGATCGC


 


26641
ATTTTCCCCC AAGTGCAAGG TAATCAATGT GGCCCTTTGA TAGCATAATT TTTAGATTAG


 


26701
CTTGTCCACT TTTATTTGTA ACAAAATGGC TTTCCCAATG TTTGATAGTT AGATCGTAAG


 


26761
TTGAATCAAT CACTGATCCA ATAGCATTGT CTGCTATATC AAACGTTTCC TCCCACACAA


 


26821
AAGAATATTC ACCGAAACCG GATTGTTGCG ATACCGAATG CTGATACAAT GTTTGATAGC


 


26881
AATTATAGTT CCAAAGTTTG ATTTTGATCC CAATATTATG TGTGTTATCT GCATCTGTAC


 


26941
CAGTTACGTT TAATATACAT CCTCTGAAAT AATACCCTTT AATCGAAATA TTACTATGAA


 


27001
ACATGGTATC AAAAGAAACC CCATACTTAG CTGCACCTCG CTCTAGCATA ATTTGAATAT


 


27061
CATTTTGATT TCCATTTCTA AATTTGAATG CAATTTGATC CACGTTTAGT GCTGGTGTTA


 


27121
ACTTTAACCT CAACGAAATA TGAGACTGCT TCAAGTCAAT AGCTAATGGA CTTTCAATTA


 


27181
GATAACATCC ATTCCCATGA ACACTTTTAT TATTATCAAC TGCATAGGTT AGAATGCTTT


 


27241
GAAGGATTTG GTAATCATCA TTTTTACCAT CTCCATGTGC TCCCCATGTT TCAGGTATAA


 


27301
TTTCATTTTC AATAAGTTCC GCTAATAGTC CTGATGCTAA TTGTTCTGCA TGTTTAGTTG


 


27361
TTGTATCTGT GACATAAAAA AGCCCTCCTC CTCCATCATT TGGTTCATAA TAGCCAAGAG


 


27421
TTCTTGCGCA CATATCTTTG CTTAAAGCAC TATCAGCTTT CATGGCAGCT ACATTTGGAT


 


27481
ACACATGTGC ATAGTTAATC GCAATTGCCT TCATCATTTC CTGCTTAAAC GCCTCTGCTT


 


27541
CCGCCTGGGT AAACAAATTG TCTTGCTTAA TCTTAGCGTC TAGTGCATCT AAACCATTTT


 


27601
GAACAGTAAC GCCAAGAGCT TTGAGCGTGT TCATTGATTG GGTAAACTTA TCAATAAAGT


 


27661
CACTGGTCAT CTTGGTGAGC TTGTCGGTAC TGTCTTGGTA CTTGGCTTCG ATTTCGTTAA


 


27721
TTAACTCTTC GACGGGTGAA ATATATGTTC GTGGTACCAA GCCATCAATG ACCTTATCAG


 


27781
CAAGGACATC GAGGTCGAAT TCCAAGGTAG TGATAGAGTT ACCGTCTTTG AGGATGCGAA


 


27841
AAAAGGCTTG GCGATAGGAA CCAGCGACTG TAAAGGCATG GCCGGGCATG TCGAAACGGA


 


27901
AACGTCCGGC GGTTGGGTCA AGGGCAACGT AGCCTTTATC ATCAATTACT CGGAAGTCGC


 


27961
CGGCGGAATT CTTAGGAAGT AATCCTTCAA ACCAGACGTT ACAGCCAGTT AAATCTAGTG


 


28021
GTGTCCCGTC TTCGTTCTTA ATATTGACAA AAACTTGACG CATACTTCGC TCATACTGAC


 


28081
GCGCTTGGAC CCAGTTAGTA TTACTGCCAT CAAAGTTAAC CTTGAAGTCT TGTACATTGT


 


28141
CAACATGCGG ACGCCGGTCT TGACCGATCA CATAAGTTAA AGTTTGTGAC ATTAAATCAT


 


28201
TCCTTTCTCT TTCAAAATCT GCATGACAGC TTGTTCAACC GTTTCATAGT CGGTCCCTAA


 


28261
AGCCACATGC TCAATTTTCT TTTTCCACGC TTCTTGAGCC TCTTTTAGTT CATCTCGCGT


 


28321
GGCTTTTTCT TGTTCGAGTT GGTCGAGACG AGTATTAATC TCCTCGAGTA GCCTAATCAA


 


28381
TTCTTTATAC TGCCTGTCAC GGAGATTATC GTCTTTGTTA TCCTCATTTT GAATTATCGT


 


28441
TAAATTATTT ATTAATTGCT GGCGGAATTT TCTTCCACTC TGCAAATCTA GATTAGTTTC


 


28501
TAAATCCAAA TGACCATCCC CCTCCTCACT TCAAACTACT CCAATCCGTC ACTCTGTAAC


 


28561
CTAGATTTAC CTGGGCCCGT GATACGTATG CTTTATCAGA TCCGGAAACA TTAAAAACCA


 


28621
GCCTTCCTTT AGTAGTGCCG GCTGGAATTT TAATGTTAAC TGTGCCGGCG GATTGCCAGG


 


28681
CGCCATTATT GACGATATAG ACAATATTCG ATTTGCCGGC AGAACTACCA TCGCTCTTCA


 


28741
AGAACTCGAC ATAAGTAGTA GCGGAACTTG AATTATCGGT CGCATTTAAC TTAGCCATGA


 


28801
GACGACTTGA GATGGAGGAG ACACCGTTGA GGTTAAATTC ATTGCTTCTG AGAGTGCCTT


 


28861
GCTCTACTTC AATTACTGGA CTGTCATTAT ATTTTTCGCT GGAAACAGTT CCGCCAGTCA


 


28921
TCATAGTTAC CAAATTATGC TGGCCATTTT GCATCCAGTA AATATTCTGG TCTACTGCTT


 


28981
CCATATTCTT ATCCGTCTCT TTCTTATCAT TCACGACTTG CGTTAATGAC TTGGATTGCT


 


29041
GATCCATGCT ATTTGCTAAT CCACTAATCC GGAGTCGTTG TTCTTCAAGA GCTTCTTGTA


 


29101
GACGTTTAGC TTTCTGATTT TGATAATCTA AAATAGTTGT TTTTGAATTA TTTAACGTTA


 


29161
TTTGATTATC TTGTCCCTTA GCTTCCGGAT ATATCGTATA ACCGACAGTC TTATAACTAC


 


29221
CTGAATATTT ATCTCGCGCA AGTACTCTTA AAATTTCACC AGCAACAGGT ACAAATGAAT


 


29281
TTCCCTGAAG CGTGACTTCA ATATTTAAAT CTGGATCAGG TTTAAGAGTA GTTAAGGCAT


 


29341
ATTTACGCAT TGCATCCGCA TCTTGAAATC TTTCATCAAC AAGATCAGGT CCAGGATGTT


 


29401
CTCCCCATTC GTCAATTGAC TTTTGATCAA CTACCATAAA CGGTTGAAAA TAATACTCTT


 


29461
CCTGTGATTC TGTGGTTGTA TCATTTCCGA CAGTATCATT GGTTTCTTTA CCACCGATAC


 


29521
CATCACGAAT AACTTGTAAG GGATCAAGCC AGGTTCCATC ATTTGACCAG GCACTTTTCA


 


29581
ATGCAGTCAT TAAATTCTTT TGCTTAGTAA TTCCAATATG AAGATGGCTA GTATCACGGT


 


29641
GGCCAATAAC ATCCCCCGTC TTAATTTCTT GGCCAACTTG AACAGTAATA TCTCCTCGAT


 


29701
TCAAAAATGC TTCTTGGTAG CAAATTAAAT AATCATTCGA CACAATAGTT ATATAGTTTT


 


29761
CCAGTCCAGC AATGTATCCA ATATCCTGGA CTTTCCCACC ATGAATCGCA TGAACATCGC


 


29821
GTCCAGGATG ATCGACAGAA CCAAAATCAA GGCCATCGTG AAAACCATTA GTTCGCCCCA


 


29881
CTCCATCTTG AGGGTGTGTT CCAAAAGTTT GCCCCAATGA AAAATGACCT TCTCCAACAT


 


29941
CTGGGAAAGG CCATCCCCAC GGATTAGGTG GTGAAATGAT CAATTTATCT TTCGTGATTG


 


30001
GGGCACCATG AGGACTCCAA CCAGTAACAC CATTAATTTG TCCTAGTGCA TTAGGAATAT


 


30061
TGAAGAACGC AATTAACTGG TCAAAACCCT TTAAGATATT TGTATAGGGC TCCCGACAAT


 


30121
AAGTATTAAA AGTTCCTCGC TTAAATTGAA GTAACCCAAG CGCCGGACCA GAACCATCGC


 


30181
CATCGGGATC AGTTCCTGGT TGCGGAATCG TCTCGTTTCC ACCTGATTCC AAATGAATTT


 


30241
GTGCCCGTAA AACATTAAGC TGCTGAGCAT TCGGTTTTAT TCCATAAAAG CTAGCAGCAT


 


30301
ACTGAATAAC TGGTGTCCAA TCACCGTTGA CTGGCTCAGT TGGTCCATTA GCATTAGTTC


 


30361
CACCTGTCCC GGTATAAACA GTGTGCTGAA CCTCATGCTT ACCACCAACA CAACGAATCA


 


30421
TATTAATAAT TGATTGACTG TCCCGAGTAG TTTTAACTGA CTTTGCATCC CTTGGAAAAT


 


30481
CAATCACTCG ACCATTATCC TTATAGAACT CATCTTCTGA ATAAACTCGT ATCTTCTTGT


 


30541
TATCCGGATA TATAACTGCA CTTGGCCATA ACTCAGTAAT CTTACTAAGC ATCTCTTTAC


 


30601
CACTGCCGCT ATCGAACTTA GAAGTTGACT GCTTATCAAA ATTACCATGA ACTTCATAGC


 


30661
TAAAGCCTAG CTTATTTCCA TCAATCCAAG CCTTTAGTAA GTCCTCAACT CCATAAGACT


 


30721
GATCCTTACT AGCATTACTA TCCTGATTGT TATTTAAGGT AAGGTCACTT CGTTTATAAA


 


30781
TTCGACTAAT CTCATTATAG ACATGCCAAG CAGTGATATT TTTCGTATTA GTATCAAAGT


 


30841
TTTCAACACA ATTTTTTACA ATATATTCCT GTCCCTCAAG CACAATCGAA CTTTCAACAT


 


30901
CTAATGGATC ATACAATGCT GAATCGTAAT CAAATACTGA AAAAGTCAAT TGAAAGGTCG


 


30961
AATTCTTAGA CCACTCGCTT TGCATTGTTG GCCAGAGAAT AATGTCTCCG ATTGGCTCCT


 


31021
TAGTTGTCTC TTTATGAGGT GTCATCAATA CTAAACTAAT TGTCCTCACC TCCTAACCAA


 


31081
GATACATAAA TGGGAAACTG AATGTAATTT CCAAATCATC TGCACCAGTA ACTTCAAAGT


 


31141
CATTCTTTCC AGGCGCTAAA GTAATATACC CATAATCGGT ATTGGCACTG TCTGGGTTAT


 


31201
TATTCTTAAA TGTTCGCCGT CCCTTTAGCA AGAGAGTATC GTTTCCAGAT AGATTTTGAT


 


31261
TGTACGTCCA AGAAGTATTA GTAGTTGTAT TCTTTACTGT GAATTTTCCA CCATTATGCT


 


31321
TAATAGTAAT TTTTAGATCA TGACGCTGTT CAACAGGATC GATTGTAATA TCACTATCGT


 


31381
TAAGGATAGA AAACTTATTC TGGCCTTTGA AATGGTATGA AGGAGTTTCC TCATCCATGT


 


31441
TCATATTCAA GTCTAAGAAA TCCTCATCTT TCATCTCATC CGTATGCAAT TTACTAAATC


 


31501
GCATTCCGCT TGGATTTTCA AAAGGCACTT CAAACGTACA ATAGTTAACT TCTTCTGGTT


 


31561
CGCTTTTAAT CTCAAAGGAA CTAGCCCGAC AGTAACGGAC AATATCCGGC TCAACTCCTG


 


31621
TTCTTAACCT AAAGATTCCT TTTTGTGCAA AAACGCGATA AATCTCATGC TTTGCCATCT


 


31681
TAAAATCTTT TCGATCAAAG AACTGCAATA AGAACTTAGC GGTGATCGTC GTTTGGCCAT


 


31741
AACGGGAATA ACTCCAAATT TGACCATCTT GTTGGGAATC ATCACGATAA TTATTAATAA


 


31801
CACTAGGCGA TTCAGTTAAG CCAAGAAAAG TTAAGTGGTC AGTAATATCT GTGCTGGCCA


 


31861
CTTCTTTTTG ATCATCAATT TTAATGTAAA GAATGTTTAC AGCGATTACG ACCACCTCCT


 


31921
AAAGATGTTG ATTATCAAAT AATGCTTGAT CAGTTCCTTC AATCCCATAA AACTGGTTCT


 


31981
TGTCAAAGGC GCCGGCTTTA ATTGCAGAAA GTTGAGCTGC ATTTAAGCCC AGCATTGTCC


 


32041
CAAAGCGGTT GAGCATCTGG TCCATCCGAT CTAACATCTC TTTTTGCATT CCATTATTAC


 


32101
TTTGGCCACC GTTATTATAA TTATTTACAA CTTGGGTGCT TTGCATATTG GGCTTAAAGT


 


32161
TAGTTAATAC CGAAACAGCA TCATCAGTAC CATTAGCATA TTGTGGTAAG TGCTTAAACA


 


32221
TCTTAGCAGT ATCGCTAGCT TTAACAACTT TTGTTCCTCG TGGTGCTGGA AAGACTACAT


 


32281
TACGCCCATA AGGAATAAAT GGTGTTTCAC CTGGGAATTG AACTAACTCA CGAAAGACGG


 


32341
GGCCAGGTTG ATCATTAACT TCCATCAATC CACCTTTATG GTAATTAGTT CCCTGAGCAT


 


32401
GCTTTGATTT ATGAAAAATA ACATTAACTG TTTTCTCAAT AGTATTAGGT ATACTCATAA


 


32461
AATTATGGAT TGCATCAAGT GCTGTTTTAA TTGGCCCAGA AGCTTTATCA TTAGCTTTTG


 


32521
CAATTTTTGC TGGTCCCGTA TTTGATCCAG CAAAAGCATT TACACCATTG CGAGCACTAT


 


32581
ACATTGGTCC ACTAGCATTA TCAACGGCTA AAGCATTTTT AGGAGCGCCA GGATTTGAAC


 


32641
TAGCAAAAAC ATCGACACCA CCAGTAGCAT TTCTCATCGG ACCACTAGCA TTGTCAATTG


 


32701
CCAGTGCATT CTTTGGTTCT CCCGGATTAG CATGACGCCA AGATTGAAGG GCATCGTTAG


 


32761
CAGCTTTTAG ATTTCCACTA GCTTTATCAT CAGCAATGAT TTGTTTTGCA GCACTAACCG


 


32821
GCATCCCCTT CCACAGCCCA TAATCAGTGA TCAGTTGCTG TAGTTCGGGT CCACCCTTAG


 


32881
CATTTACGAT TGCTTGCTGT TGCTTAGGAC TAAGTTGATT CCAGCGACCA ATTTTATTGA


 


32941
GCGCATCCAT TAATCGACCA GCACCCTGAG TAGTAACCAA AGCACGTTGT TGCTTAAACG


 


33001
AAAGACTATT CCAAACACCA GCCTGCACTA ATGCATCAAC TAATTGTGGA GTTCCTTTAG


 


33061
CGTCAATAAT CGCTTCCTGT TGTTTTAAGG TTAAATTATT CCACTGTCCG GAACTTTGCA


 


33121
AGGCTTCATA AATTGGCTTA GTTGCCTTAT TAGTAATTAC TGCTTCCTGT TGTTTTAATG


 


33181
ACAAGGAATT CCATGCACCG GATTCAAGCA GAATGTCGGC CATTTCTTGC TTACCTTTAG


 


33241
CCTTAACAAT TGCTTCTTTG GTCTTAAGAT CAAGATTATT CCATTCTCCC GACTTCTCTA


 


33301
AAGCTTGAAC AATTGTTTCA CTAAAGCCGT CTTTCAACCA AGCTTTCTGC TCTTTCCAAC


 


33361
TCATGCTATC CCATTTACCA TTTTCGATTA ATGCAGCTGC CACCATTTGT TGAGCATTGG


 


33421
TACTTAACTT ACCTTCCTTT TTCAGAAGTT TAATTTGGTT CCATTGATCT TTAGAATTAA


 


33481
CAGCCTTATT AACTTCCTCC TGGGCATTAG TCCGAACTTT ACCAGTTTTG GGATCGAAAA


 


33541
CTAAGTCATT CCACATATCA GCGGCTTTTT TAGCTTTTCC ACTTAATTTA GTAGTATTTA


 


33601
CCGCTAAGCT TTGTAGGTTT GTTTCGGCAG CTTTGGATTG CCGCTTAATC TCATCAGTGC


 


33661
CATCTTTATA ACTAAGTCCC ATACTTTTTA GGTCATTCTT AATTAATTTA GTACTTTCAC


 


33721
CATTAGCACG GGCCAGGCGA ATATATTCGG CAGCTGCTTT ATTCGTGTAA TCTGTTAATG


 


33781
CTTTCTTATT AGCACGCATA CCGACCTTAT ACAGTTCAGA ATCTTTTCCG TAGCGTTTAG


 


33841
CAAGCATTGA AGACTGCTTC TTGTATTGGT TTTCCATTTC AGCAGTTTCA CTGGTTATAT


 


33901
CTCCCATTTT TGTTGAGCGT TGTTGCATAG ACATATGCTT AATATCGTTA TTAAGGACAG


 


33961
CCATTGCTTT AGCACGCTTA CTTCCACTTA ATTTAAGAAT TTTCATCTCA TCGTTAAGCA


 


34021
TTTCCTGCTG GTTGTTCTTT AACATGACTC GCTGAGTATC ATTTAATTGT GAAACTTTTT


 


34081
TATTGTGGTC AGATAGAATT GTTTGAACAC GCTGATTGGC ATTATCTGCA TCATTCACAT


 


34141
ATGAATTATA TTGTTTTTTC TCCTTTTCTA ACTCATCGTC AATAGCTTTA GATATTTCTG


 


34201
GAGACAGACC TTTTTCTGCC TTTTTGGCAT TAGCAAAATG TTGTTTTGCA CTATTTTCAA


 


34261
TATTAGTAAA CTCACGGTCA AAATCAGCAG CTAATTTCTT TGTTGAAGTT GAGGCAGCAA


 


34321
CATCCATATC TGACAGAGTG TTCTGAATTC CTTGAGAAGT CGACTGCATT TTTGATAAAG


 


34381
CCTTATCAGC AGCTGCACCA ACATCAGATC CCCAACGATT CGTTCGTTGT GATGATTCGA


 


34441
CAGCTTTCTT ACCCCAAAGC TCCCAAACAG TAATTCCACC GGCAATGGCT AAAGTAGCAA


 


34501
CTCCAGCAAC AGCGGCAACT GTTCCTAAAG ATACTCCAGC CGCAGCAGTA GCAGCACCTG


 


34561
CTTCACCTGC AGCTACACCG GTACCTTGTA AAGCGACAGT CGCAGCACCA ACACCGTTAG


 


34621
CAGCTTGAAC CGCCCGACCA CCTGCACCAG TCATGGTTGT TCCAAACTTT GCAGCTTGAA


 


34681
ATGTTGATTT TGAAAAAGCT GAGCCAATAA CGTCTAACCC ACTGGCTCCT AGTTTCATTG


 


34741
CAGTTTGTAC ACGGCCGATT CCGGAAGCAA TTTTTCCAAA GGCAAGAACT GTCTTCCCAG


 


34801
CACCACTAGT CAGTTTACCC AATGCTAGGA ACAACGGGCC TGCACCAGCA GTGAACAGGG


 


34861
TAGTTGTTAC AATTGCCTTC TGGACAGCTG GTGATAAATC ACCAAAGCCA TGTGCAAGTC


 


34921
TGGAAAGGCC TTGAACCATT GGAATAATAG ATGGTAAGAC GTACTTTGCC ATATCCATTC


 


34981
CAGCGTTAGT TAAAGATTCC TTGAAGATTG CTAATTGTGC TTTAGGAGAT TGAAGGTTTT


 


35041
TCTGTGACAA GTCGCCGATA TAATCCCGCT TGGCAGAGTT TTGAACTTCT TTATTAAGTT


 


35101
CACGTAACCG ATTAGCGTTT TCGGTTAAAA TTGCCCCAGC TTGTTGTCCA GTTGTTCCAA


 


35161
ATAACGCATG AAAAATATCA TTCTTTTGGT GACCAGATAA GCCTTTCATA TGGTCATTTA


 


35221
ACGTCTTAAA AATAGACGAC ATTGATTTTA ACTTTCCACT CTTAGTAAGA AAATCTTTAG


 


35281
TACTTAAATT AATACTTGCT AAAGCTTTTT GACCATTAGC GGTTGGCGTA ATTAGTGAAT


 


35341
TGATAACCTT TCGTAGACCG GTACCAGCTT TGTCTGCTTC TAGACCATTA TTAGACAAGA


 


35401
TCCCCATTGC ACTAGCAGTT TCTGAGAGAC TAAAACCTGC TTGATGAGCA GTTGAGCCAA


 


35461
CATATGACAT TCCCACACCA AGTGATTGAA AATCAGTTGA TGTGGCATCG GCAGCATAGG


 


35521
CTAACTCATT TAATGTTTTA GTAGATCTAC GCTGCATGAC TGTAGCATTC TTGATTGGAC


 


35581
GTCCTGCTTT ATCAGTAGCC AGTCCAAAAG ATTCCATTGT CTGCGAAGCA ACTTTGATTA


 


35641
CATCATTAAA GTCATCCCCA GTGGCCACAG ATGCTTTAAG TTCGTTCCGC ATTACTCCAA


 


35701
TTGCTGCTTT AGATGTATAA CCACGCTTTA CTAGATCTTG ATACCCGGCA GCAATCTTTT


 


35761
GCTGACTAAC ACCGTACTCG TCAGAATACT TTCGGGCATC TGCAGTCATC GTTTTATATG


 


35821
CGGCATTCGT TTCTGCAGCT GATTCTCCGG AAGTTCGAAT AACGTTCTTG ACCTTTGTCA


 


35881
TTTGATCTTG GAAGTCAACG AGTTTCTTAG CGGAATATGT TAACCCTGCC GCAATTGGAG


 


35941
CTGTTAAATA AGTCGACATC CCTCGACCGA CACCTGATAT TTTAGAACCA ACATTGGTGG


 


36001
CCACATTTCC AAAATGTTGA GTGCGATTAG CAAGTTGTGT CCACTTATTC GACTGCAATT


 


36061
CAATATCACG ATTAAGAGCT TGCATTCGTC CACGTAATTG TTCAATTTGC GCAGACGTTT


 


36121
TATTATATTG GGTTGCTGCC TTAGTTCGTG ACTGCTCGCT CCTGGTAGTA TCATCCATTG


 


36181
CTTTCTTAGC ATTCTGGAGT TGCGCATTAT AATTGCGCAT TTGTTGACTC ATAGTCGCAT


 


36241
AAGCAGCACG CATATTATTA ATACTTCCAC CGGATGCTTT CAGTGCCGCT TCTTGCGCAC


 


36301
GCAAAGCATT GGCTGTTGAT TTAATTTGCG CTTTTAAAAC ACCATTAGCC GCTTTAAACG


 


36361
GATCAATATC CAAACTAACA GTTGCTGCCA AATGTCCTAA TGATTGGGTC ATTATTTAAC


 


36421
CTCCTTTCCT AGAATAAGAA TGGGAATGCT TTATCAATGG TTGTCTGCTT TTCTTCAAAG


 


36481
ACGTAATTCA TTAATTTCAA ATCTGAAAGT GTTAGCTTAC TGACCTCATT CCATTTATAT


 


36541
CCATCGCGCA TTTTATTTTT AATGAAATCT GTGAGACTCT TGATTGAATC ATCAATCATC


 


36601
TCAACAGTTA TTTTTTTGGC TTTTCGTTAT CCTTTTTCTC TTCTAAGTCA TCTTCACTTA


 


36661
GAGGAGAACC GAGAGTTTCA TTAATTGCAT TAACAATGCT AATTAACCCT TCTATAGCCG


 


36721
TAACTCCTGA AAGAACATCC TTTTTAGTCA ATCCATTCTT CCAAAAATTA GCAGCAAACT


 


36781
CAGCCCGTAA ACTTAATAAT CGTTCATTAT CTTTATCTGT TGGCAATTTT TTGGGATCAG


 


36841
AGTACATCAC TATTTCTTGA CGTTGAACCT TTAATGCATC TAATAGATTT TCAAGCATTG


 


36901
GGGCTTCTTT TCGTTCATAG ATTGTTTCTT TGCCATCTGT TTTTACTTTT AATTTATATG


 


36961
GCATTTCCAT ATCCTCCTCA TCGTCTCACT TAACTCGTCT CTGTCTGATT AATTAGTTAC


 


37021
CCTTTATCAA CAACAGTTTC TGGTTTAGTA TCTTTTGTAT TGGTTGGATC AGTTACTGGA


 


37081
GCTTCTTCCC CAAAGACCAT TGCATGGAAC TTAGCAAAAT CAAATCCTTC ATTATCTTCC


 


37141
CGACCGATTA AAAGGATGGT TCCAGTATCC GCATCCCCAC GCGGAACGAA ATTACCTTCA


 


37201
ATTTCATCTG CTTCTGGGTC TGGAGCACCG TCCTGAGTCT TGATTGAAAT ACCTGGTAAG


 


37261
GAGAACATCC CCTTGGTTAA ACCAACCCAA CAATGCTTCC CGTTTGAAAG CTTTGTCCGG


 


37321
AACATAGTAG CAACATAATT TGGTACAAGA TTCTTAGTAT AAACTTCTGT ACCGTTCTGA


 


37381
ATATCAATTC CATACAAGTC TTTCTTCATG ATGGAATCAA TATCATAAAG GTTGATGGTT


 


37441
TCCTTAGCTT CGGTAATTCC ACCAGAAAGA ACCAAGTAAG GGCCGTCATC GGCTGCTAAC


 


37501
GTCTTTAATT CGTTAGTCAG CTCCAGCTTA ACTTCACTTA ACCCGCTCAT CTTTCGGGTA


 


37561
TCTTTAATCT TTTCGTTCTC GTCTAGCACC CCATATTCAA AATTAGAGGC CCCAAACTTA


 


37621
GCAACTTTTG CATTTGGTGT TCCCATTAAA TATCATTCCT TTCTTCAAAC CCTTCAAAGT


 


37681
TAGCTGTGAC CATTATGCAA TTATCCAAGT CCGGATCCGG ATAAGAATTC TTGTAGTAAC


 


37741
GTTCAAAGCC ATGACTGTGG AGGGTTTCAT ATATTCGTTC TTGAATATCC ATTAGCCGGT


 


37801
CATCATTCTC TTCACGAATC CAAAAATCAA CTTGTACACG TGGGTATTCA AAAAAGCGAG


 


37861
CATTGTCACT GTAAATTGCA TCATCCCCAG GTAGGGGGGT GATCCGAATC CACGGTGCTG


 


37921
AACTCGCTTT AATAAATGGA TCATCAGGCG TACTAGTAAA AATTGGAACA TATGATAATT


 


37981
TATCTTGTCG CAACTCATCC ATCATCACGA CTAGGCACTC ATCATTCGTT AGATAATCGG


 


38041
CTACTTGCAT CTCTGGAGTT TTCATACTTT CAAGTCCTCA ATAAATTTAG CCAATATTTT


 


38101
CTTCTTAGCA GCATTTCTTG AATCTTCAAC AAAGTGTTGT GGATCTTGCT TAGCAGTTCC


 


38161
AGCATCGGGG AAGTGAGCAA TCCATCCTTT ATCTTTGTCA TAACCAACAT CGACAGAATA


 


38221
ATCTCCACCG TTAGTCTTGA GATTACCGTG TTTAGTATGA TCTTTTAAAG GAGTCTTGCC


 


38281
TGAATGATCA GAGCCTTTAG AAGCCACAGG CGTCCATTTT CTTAATTCTT CTTGAAAGAC


 


38341
TTTAGCACCA TCACGGGTTG CTTTTCGTGC CTTCTTTTCT TCCGTCTTTT CAAGTTTAGT


 


38401
TAAGTTAGCA ATCAGCTCTG CTTCACCGGT CACTGCCATT TTGGACAACC TCCTGGGCGG


 


38461
TAATTTTTGT TAGATCTCGC TTTGCATAAT CAGGATCCAT TCCCGTGATT TCATACCATT


 


38521
TTCCTCGCCA GTTAATTAAC CAATTCGATT GAATTTCCTT ACGAGTTTTG AAAGCAATTA


 


38581
AAAACGTGGG TGATTCTTTT CTGAACCCCA CGTTTGAACT GTTTTGGACA AATTCACGGA


 


38641
CTGACAGCTT AGGAACTTCT GCCCACACGG TAAATTCCTT AACTTTCTGA TCCTTGATCG


 


38701
GTCGATGAGT TTCCGGATTG ATCCCCATTT GGACTGAGTA AAATGTTATC CGTTCCGTCA


 


38761
TGTTCCGTAG TTTCATCGGT ATTCACCTCA CTACTTATTT GATTAATAAG GCCATCAATT


 


38821
CCTGATGAAA GAACTGGTCG GTAACTATCT GCAGTAATTC CTCGCTCGTA GAAGTCTTCT


 


38881
TTCACCTGCT TCATAAGTGC AATTTTGAAT CGCGGTTCGT CAGAGTAGTC AGCAGGCTTA


 


38941
CTCTCCCATT TAATTGCCCT TGCAATCATC AATGCGGCAG CATCAACAAT TATTTTAAGA


 


39001
ATTTCATCAT CAAAGTCCTG GTCAATTTTG CAGTAGTTTT TTAGATTGGC AAAGAATTGT


 


39061
TCATCACTCG AGAAAGTGTG GTCTGTTTCC ATCCTACTCA CCTAGCTTTG CTAACAGTTG


 


39121
ATCCTTAGTA TCGGTTGAAG TATAGGCAAT TCCATGATCA TCTAAGTACT TTTTGATTGT


 


39181
GTCTACCGTA TCAGCACTAG TCGGTTTAAC GTCCGCCCCG TTGTCGGACG GCGTTATTTT


 


39241
GACGCTGCTG ATGGACCAGT AACAAAGTAA CCAGCGTTTT CATCGGCCTT CTTCACATCT


 


39301
AAACGAAGAA CCGCTTGGAG ATATTGACCA TAAATGTCAT TATCAACCCA CCGTACCTGG


 


39361
AGATCTTTTC GATTTGCTAA AATAATTGCA CGGTATGGAT CACCAACAAA GGCATGAGCT


 


39421
TCACCCTTTG CTCCAAATAA GTCATCTTCA ATTACAGCAA CGTTGATTCC AGAGACCGCT


 


39481
TTACCAGACG GACTAGAAAT ACTATCTTGA AGTAAGTAAC GTCCATTCTT ATCTTTCAAT


 


39541
GTATCAAGCC AGTTGTAGAA GCTTTGGCTG GCAATAATCA TCTTATTGTA AGCCACATCT


 


39601
AAATCGACAT TCCAAATTTT CTTTAGGTCA TCGATTGCAT TAGAGCCATC AACGGTCTTA


 


39661
GCTGAAAATC CCTTAAAAAT TGTAGCAATA GCCGCATTCT TTGTATTTAA GGATTGTTCT


 


39721
TTTGCGTTTC GAGCAACTAA TCCAGTTAAA TCAATTGCTG AATCATCAAT CGATTCTTGT


 


39781
GAAATAGGAA TTGCTCCACG ATAGGTCTCA ATTTGCCATG CTACCTTAAG AAATTCAGGT


 


39841
TTTTGAAGAT CAGGATTCTT TGCAAGTTCT TCAACAGTAT GCATTTGAGC AGTTGCCTTT


 


39901
TTAAGAATTG GATAACTACC AGAAGCAGTT GTGGCGTTAA ATACTTGAAC AAATTGGCTA


 


39961
AGATCAGTTA CTGTTTTAAT TTCATTTTCA GGATTGTAAA TGATTGATTC AGGAATAGTA


 


40021
ACGCTTGCGT CTGAAGAAGT AATTCCAGTA GTACCATCAC GATGTTCTTG GTGAAGATAT


 


40081
GCGTTGAAAT TACGTTTTTC TTCATTTTCG TCCTCGTTAT TTTGAGAACG CTTATGAGGA


 


40141
TCTGGAGCTG GATTACCCTT AGCTGCTTTA CGGTAAAGCT TAAGGTCATC TTCAATACTC


 


40201
CGAACTTCCT TTTCAGCAGC TTCAATTTCA GAACGTAATG ACTTAGCTCG TGTTAAGTCT


 


40261
TCATCAGTTG CGTCTTCATT TGAGAGCAGT TGGCGCATTT CATTAGTTTT TTCGTTAATT


 


40321
AATGCTCGCT TACCTTCTTT TTGAGCAAGC AATTCTTTAA TTTTTTCGCG AAACATTTAA


 


40381
TTTCCTCCTT CGAGTGTCTT CAATAATTCT TGTCGTTCTA ATTCACGTAA CATCTTTTTA


 


40441
CGTTTCTGAT CAGTAGCATC TCTATTTTCT TGGCTCTGCA TTTGCTTAAC CATATTGATC


 


40501
GAACGCTGAC CGACCTGAAC TTCTGTATCC GGATAAGCAG GTGTTGTAAC TACTGATACA


 


40561
TCGAACAAAC GATCAATTTG TCGAATGGTT CGTTCATAAT CAACTCCATC CTGGTCAGAT


 


40621
TCTTCCCAAT CTTGGGCCTC ATCTGTGTTG GCCACCGTAA AAGCAAAGCT GCATTGATTA


 


40681
ATAACGCCTG CCTTGATATT GGCGATCAAA TCACGCGCAA ATGATGTATC AGTTGGCTTA


 


40741
ACAGTGAATT TCAAGCCAAT TGCATCAGAA GATAAAGTCA TGTTCACCCC TGATCGTCCC


 


40801
AGTACCTGGT TAGGATCGTG ATTGATCGTC GCTACTACAT TTGACATATC AGCGTCATCG


 


40861
AACGCTCCTG GAGCAATTTG CTCAATAAAG CGAGTAAAGC CACCCAAAAC TTCGGATGGC


 


40921
TTATTGTATT TGGCTGCATA ACCTTCAATT ACTGGTTCAT CATCTTCATC AGTTGCCGTT


 


40981
CTCATCTGAA TTGGCATCAT TAACTGTCGA GTTTCCAAGT CGCTTGTCAT CGATTCCACC


 


41041
TCCCTTCGCT GTTGATTGTT TCTGATACTC TTCTTTTTTA TCAAGGAAGA CAGTATTCAA


 


41101
GGTTGATTGG AAACGGTCTA AGTCTGGATT GTCAGATTTG ACTAACCCCA TCCGAACTCG


 


41161
TCCTTCATTA GGAGTAATGA CGTTATTTGT AACTCCCTTT TGTACATCGT CCATTGACAT


 


41221
CCCTGTTTCT TTGCGTGTAT CAAATTCGAT GTGGCAGTTA TGACGTTGCC GATCAGTCAG


 


41281
CATTGTCATT TCCAAATTAC TTGCAATCGG CTTGAAGTAA TAAGGTAAAT CAGAAGTAAT


 


41341
AAAGTCTTCA TTTAATTGTT TAATCGATTG GTTAGGACTA TTAACTGCTA ATTTATATGC


 


41401
TGGAATATGC AACGCTTTGG CAATTTGAGC AGTTGAATAG TTGTTGCTGT TAATTAGTTG


 


41461
CAAAACATTG GTATCAATCT CAATTGGCGA GTAATCGAAT GTATCATCAG TAACGATTGG


 


41521
GCTACCGGCA TTGCTATTAG CTTGTGCATA TTCAAACGCC TTTCTAGTTT TAAGACGCGC


 


41581
TTCCGGACTT AACTTACCTT TAGCCTTTAG TAATCCACCT TTAAGACCTG ACTTAAAGAA


 


41641
CCGCCGTAAA GTCTTAATTC CATCATCCTG TAATCCAATC TCATCAGCCA AAGACAACAA


 


41701
CGGCGACCGG CCATGAATAC CATCGTAGGT AAAGAACATA AAATGAATTA CATCTTCAGC


 


41761
CGGCACAACG ATTGTTTGAC CACCGCCTTT TTGATTGATA GGTGTAAATT CGTATTTAAT


 


41821
ATTCGTTACA TCAGAATCAT CAATATAGGT TTGCGATGGT GGAAAATATT GTATTTCTAA


 


41881
CGGTGCTTTT GTATGAGGAT CACGAATTAT TCTTGAAAAA CCATCCCCAG TTAAAATTGC


 


41941
GTTAACTGTC ATTAGGAAAC GCCAATGATA AGCAGATAAC ATATCGTTTG GGTGCTTATT


 


42001
CAGCAAATAA TCAACACTTT TGATATTCTT TACTGCATTT TTACCATCAT CCAAAATGAC


 


42061
AATCGGAAAT CGAGCTACAT TACTAGCAAC ATGAGAGACT GCAGTCAGCA CATCAGAGTT


 


42121
TCTAAGAGCA CCGATTCCAC TATAAGATGG CATATTGCTA AACCCCGGAA GAATCCCCTG


 


42181
GTCAATATAA TCTTGTGCCC ATTCGCGTTT CTCGGTATGA AATAACACTC AGCTTCACCC


 


42241
CCTTTCATAA GAGATAAAAG CCATTACGAA TAATTCAATT GAAATAATTA GTAAGCCAAT


 


42301
CTTCGTGGAA AATAAAAAGC CTGTAACTGC TAAACAGATA CAAGCTAATA TGAACAGCAA


 


42361
AACCGGTTCG TTTAGTTTCC AGAATTTCAT GGCATCACCC GGAGAAGAAA TTTCAAGTCT


 


42421
TCATCCGTAA TCTCAAATTT CTCAATATAC GGGGCAAGCT TTCTCAGTCG TTCCAATCGT


 


42481
CCCTTGATTT CATTAAGTGC GGGAGTTACA TCAATCTTCA TTCCTTCTTC ACCGTCGTAT


 


42541
AGATACTTAG CCATGTCTAT TCCTCCTCGA TTCCTTTGAT AAATTCAATT GCTTCCTTAG


 


42601
TTAAGGCCAA TGGATGGAAA TATTCTCCAT CTGAACAGTC ATTGATGATC TTCCCTTTTA


 


42661
ATAGGTGCTC CACATCATTT TTAGTGATAA AGGTGGTCCA GTTGCTATCG TTGCCATATT


 


42721
CATCTAGTTG CTTAATATCA TGGCTAAATT TATCTTTTTC AGAAACCAAA GTCATCATTA


 


42781
AACACATCCT CATCTGTTAA ATAATCATCA ATATTTTCAC GGAAACAAAT TGCATATGCG


 


42841
TCTAGCAATG CATCGGCAGC ATCAATCTTA TTTGAATAAC GGTTCTTATC AATCCGGACA


 


42901
CCATTGTTAT CTGATTTAAG GATCGCGTTA GCCATTGCGC CAGTTAAAAT TTCATTGTCA


 


42961
GAATGTCGAA CTCGCTTATC TAAAATATCA TCCCTAAATT GCTTGGTTGG CATTGAAAGA


 


43021
GTTAGTGTTC CTTGACGAAC AGATATCTGT TGCCATTCCG GATGACCTTT TTCAATCTGA


 


43081
GTTAACAAGG TTCCATATTG TGCGGGGTCA TAACAAATTG CCTGAACATC AAGATTGTGT


 


43141
TCTCTGACAA AGCCATCCAG CCATTCATAT ACCCGTTCAA CATCAATAAC TCCAGATTCA


 


43201
AGCTTCGTAA TTTCACACTG GCCCATCCCT TGCAACCGGA CATAATCTAA CCGGTCTGCT


 


43261
TTAATCTTTG CTTCCAAACC ATATTTAGTA CCGACAAACG CATAAGAATC CGCATACCAA


 


43321
TACCCCTCTT GAGGAATTAA CCAACTAATT GCATAAAGGT CAGATGATTT ACCAACGTCA


 


43381
ATCCCAAACC AAACTCGTTG ACCATCAATG TCAATTGGAT CAATTTGAGC TGCATTCCAA


 


43441
GTATCAATAT CCATATAACT ATCCTCTTCG GCTTGTCGCC ACATATTAAA GTTCTTAACT


 


43501
AGAACAGAAT TCTTTGTCCC TTTCTGTTTA GCTTCTTTCC ACCGCTTTGC CAAATATCCA


 


43561
TATACCTGGT CTTGCAAAGC CGGGACACTC AGAATTGGAT TGGATTTAAT CCAGGTACTT


 


43621
TTATCATCAA CTTCTGATAC ATTATCTTGT TCAGCAATGT AGGCAAAATA AGTATCATCA


 


43681
GTAATTTCAC CTTTCAAAAC CTTCGTTGCA TAGGGATATT CAATTGTATG CATTGGAACA


 


43741
TTCAAATCAA AGCCGGCTGT TGAAATAATC ATGATAAGTG AGTTATGAAG TAAGGCCTGA


 


43801
CCGGATTCTA GTAATTCCAT CATTTCAGTG GTCTTACTTG CAGCATACTC GTCTAGGATT


 


43861
CCAACATGAG GTTCAAAACC ATCAACCGTT CCCGTTTCCT TGGAAAGAGA CCGAACATAA


 


43921
GAATAATCAT CAAGGTTACT AATTAAATCA CGATTAACTT TTGTCCCACG TTTTGTATCA


 


43981
CCATCACTGG AACGTAGAGC ATTGAGGCGT TTCTTAATCA TATTAAAAAC GATGTTCGCT


 


44041
TGCTTTTTAT CGTTAGCGGT ACAAAATATT TGCCGGGAGA ATTCAGGTGA GTTACCCATC


 


44101
AAGAACTCAT ACAATGCAAT TCCGGAAATA AGGATCGACT TACCATTTTT TCGTGCCATT


 


44161
GATAACATTC CTTTTCGGAA GCGCCGTTCA GATGGCTTGT CTTTTTTCCA CCAGCCATAC


 


44221
ATATTAGCAA TAATGAAACG CTGAAAATCT GCTAAAGGAT ACGCCCGCAT CGTTTTTGGA


 


44281
TCCGGAAGTA TTTCCATGAA TTGAATAACC TTATTAGCAC GTTCATTATC ATAAAAATAT


 


44341
TCAAAGTCAT CGTTATCCGC CTTTTTAAGA TCATTTAAGT ACCTTTTTGC AGCTAAAATC


 


44401
ACCTTTTTAC CAGCAACTAT TTTGCCGGCC ACGACCTTTT CAGCATAATC TTTAGCGTAA


 


44461
TTCATCATGA AATCCTAAAC TTATCACGCA AGGATTGATG CTCTTCCTTG TCAGTTTGTG


 


44521
GCATGTTCAT CTGCAGCCGC GAATTAACAT TAAGCCCAAG ATCAGATGCA AGACCCTTTA


 


44581
TTGCCCTGGT TGCTTTATCA ATTGTCTTAA TTGCGTTATT GATTTTAGCA ATGTTCTTTC


 


44641
TTTTCTTGGA CTGTTCCTTA TTTAATTGAA CGGAAGCATC TTTATAAATG CTGTACCAGG


 


44701
TACAATACAA TTCCAATTCG CTCCGGTCCA AGTTACGCAA TGGCAGCTTT CCGATAGAAC


 


44761
CAATAATTCG CCGATATTCC GCTTTGGCCA CCTTATCTAA ATGTGCCGGC GGAGTCTTTT


 


44821
GCAACTCAGG TAAACCATCG GCAGCCATAA ATTCTGCCTT ATATTTTGCT TCTTGTTCAA


 


44881
CCACTCTAAG GTGGCCTGTT GACTGCGATA ATAATTTTTG CTTTCTTGCC AA.






Bacteriophage loci 3 has a sequence of:









(SEQ ID NO: 12)










1
CTATACAAGC GCCCGCAATT GTTGAATCAT GCTTAACACT TGGTAAGGGG TATTTGACAT



 


61
GTCAATACCT CTATTTTGAT ACCAAAATTG CGCCAACATC TGAACAGCGA ACGTATATTG


 


121
TTCATACTGG GTTAAGTCTG CTGTGGGGTT TACAGCGTTT TGTACATAAG CCTTAGCGTT


 


181
CTTTAGATAA TCCTTGATTA ATTGGTCTTC TTCTTCACCG TCAAGATATA AGGCTTGCTT


 


241
AAAGTTGTCT ACTTCCCACT GCTCTACATC AAATTTCATT AATTTCACCT TCTTTTAGAA


 


301
CACATCTAGG TCTAACTTTT TAGCAACATC TACTATAAAT TGATCACGTA ACTTGTACGC


 


361
TTTTGTAGTG CTACACCATA TATCGCCAGT TGCTACTAGG TTCTTTATCG TCTTATGATT


 


421
AGGGTTAAAA TAGCATTCTT TAATTATCTT CTGGGTACAG TCCCCAGCGT CTTGTAATGC


 


481
TAAGTCAATA GCTTCTTTCT CACGTTTTAA GCGGTGCATA TATCTATCTT GGTCAATGGT


 


541
TATTAACATA TCATCGTAAT AAGTCCATGG TCGATTGATA GCACGTCCAC CACCTACATT


 


601
TTCATCTCTT TTGCGGATAG GGTGCATTAG TTCAAACTTA CGGCTTTTTA GTTTCTTATT


 


661
AATTGAAGGA TAATCTTTCA GTATTACTAT TACTAGACTT TTTACTTCTT TTCGCATTAA


 


721
TACACCTCAT AAAAAAAGAG TGGGCGATAT TGCCCACCCT GTGCTAGTAG CTATTATTTA


 


781
CTTTCGGTCG CCTTAGTTGT GGTTAAAGAG AGATTAAAAG CAGCGTCATT ACTAATTGGT


 


841
TGGTAGTCAT TACGGACGAT AACAGATAAA CCTTGACTGT AACTGTCGAA CTTGTCCCAT


 


901
TGGGCTGTAA CTTGATTACG CCGGAATACT GCTACCGCTT CTGAAATATC ACCTAAAATC


 


961
ATTGGATGTG AACCGTCTGT GTTATCTGGT AAGAACTTAT TACTAATCTT AACAACGGGT


 


1021
TGGCCAAGTA AAGAAAATCC ACTAGGAGCG GTTACATCTG GTTGTAATAA GTAACGTCCT


 


1081
TCATTGTCTT TCAAAGTATC TAATACTTGG AAAGCTGATT GATTTACTAA CCATGTAGAA


 


1141
GTACTTTGTA AGGCTGGGTC AAGGTCAACG TTGAATACTT GCTTCAAGTC GTCAACATTA


 


1201
GCTACGGTTT TCTTAGTAAA GTTATCGCCT TGTAATACCT TGATAATTTC AGCGTTATCA


 


1261
GTGTTATCAA CTAACTTTTG AAGTTGGTTC TTTACTTCAG ATACAATATC AACTTCTGAA


 


1321
TCGTCAACAA CTTCATTTGA TAGTGCAATC TTACCGGCAC GGGTCTTTAC ATCAAACTTC


 


1381
ACATCTTCAA ACATATTAGC GTCTACATCA GCAATTTCTG CTTGTTCTTC CTTAGTAGTA


 


1441
AGTACCGCTG TGTTGTAACG GGTAGCGATT GGATAATGAC CGGAACCAGT CCCTACAGTC


 


1501
TTTACAGTAG CGTACTTTGC TAAGTTATAA GCACTGTTCT TTAAGTCTAG GACTGGAGTA


 


1561
ACAACTTCTT CTGGAATAAC GGCTTCTGCC CCCTTAGTTG TTAATCCGTC ACGGGTTTCA


 


1621
CCACGTGAAC GAATGAAGCT TTCAAAGGCG TTAGCTGGTT GTTCTGTATT GTTTTTGGGG


 


1681
TCTAAGATTG TGTGTTGCAT TTTATTATGT TCCTCACTTT CGATAAATTG GTTGTAACTA


 


1741
CGAATGCTTG TTTTATCTAC ATCAACATTT GTCTGATCAT ATGAAGGCAC TGTAACGGTT


 


1801
GAAACCTCAA ATAAATCTTT AATCTGATTA ACGGTGCGTG TAATTTGTCC GTTGTCGTCA


 


1861
CGTGTCCAAC TGTCAGAACC ATCATCAACA TCAAAACGGA AAGACATAGA ATTAATGTTC


 


1921
CCATTCTTAA CATTATTAAG AGTGTCACTT GCGTAACTTA CACTTGGGTC AATAGTTGCC


 


1981
TTGAAGTGAA GCCCCTTATC GTCCACATCA AGTTGTAATG AACCAGCCTT AACGCTTGCT


 


2041
AGTGGCTTAG TGTAGTCGTG TTGATCAAGC ATAACTACAT TGGATAGGTC TACGTTATCA


 


2101
AGGGCGCTAG GGTCAATCAC TTCAACAAAA CCGCCTAAGT CCTTGCTGGG TTGATTAAAG


 


2161
AGTAACGCAT AGCCTTCAAC TGTATTAGGC TGTGAGGCTT GCGTTTCATC TTGGTTATCT


 


2221
TGCTGGTCTT TTGTTGGGTC TTCTTTAGGG CTTACCGTTT GTAAACCTGC GTTAGTTGTT


 


2281
AGTCGTTTCT CCATTGGCTG TAACTCCTCC TTCTTTTTCA ATAAAATTAT CTCCATTAGT


 


2341
TACCGGCACT AAATTAAGTA AGGCTCGGGC TTCATTGCGT GTGTAGATAC CGTTAACATA


 


2401
ACCATCAACT GCCATCTGCT GTTGAGCGGA CGGGTCAAGG CTTAATAGCT TGCTATTATC


 


2461
AAAGCTAAAA TCTTGCCCTA GCTTGTAGTT CAATTCAGAA GTAAAGCAAT CTTCATAGTG


 


2521
CTGTAAAGTA CCTTGTAAAT ACTGAACGTT ACTTTGTTGT TGGTTGCTAT GCTCGTTTTC


 


2581
TACACCTAAA CGCTCGGGTG GTAGTCCGAA CGCTTTCGCA ATTTGACGGG TATTCCAGTC


 


2641
ATTAGAATTT ACTAGCTTTA ACACGTCTGT ATTAAGTGGT AGGCTTTTAA CGTCCATACC


 


2701
ATCATCAGTA ACAATTGTGT TAAGTGAATT ATTGCCGGTA GTTGCCTCAT CAAATTGTTT


 


2761
TCTAATGTTC TCTTTACCAT CCGAACTAAG ATCAGCTTGG TGGACTTGGA CAACTGTTGT


 


2821
GCCATGAATT CCATTATTAA AGAATCCAGT CATTAACCTG TTTCCAGCCG CCTGTATCTT


 


2881
CATTTCATCT TTTAGCGCAT ACAGTGGCGA TAGTCCGACT AAACCATCAC GGGTAAAATA


 


2941
CTTAAAATGT AAGATGTTAG AAGAGGCAAT TACTCGCTTA ATACGTCCAT CTGGGCTGTA


 


3001
TGTGTAAGTC AGCTTTTGCG TTACATCATC TTGCTGGACA GTTAATTTAT TGTTAGGAAT


 


3061
AAAATTTAGC GTGTGTGAAC TTAAATCAAT TTCGGCAAAC GCATTACCAT TTAATAGCAT


 


3121
TTCAACCGCT AGGGCGTACT TGAAACTTCT GCCGTCCATT AACTCGTTAG GATGATCATT


 


3181
AATCATCTTG TTATATAGGC TAGAACTAGA CTTAATCGGA TTACTGGCAA GATCACCAGC


 


3241
GATAGAACTA ATTGCAGCAA ATATATCACT ATTACGTAAA GCCGAAGCAC TAACGTAACT


 


3301
ATTAGGGTCA TTACTAGAAA GACTTACTAG AGCGTCTAGA AAGGGGTCTT GTGCGCTTGT


 


3361
AGTCGTTGTT CTCTTACTAA ATAGGCCCAT TATCTACCCT CCTTTACTTG TGATTATCAG


 


3421
AATAGTTAAT CATGAAAGCT AGAACAATCA TTTCAATACC AAGAAAGCCA ATCCCAAACA


 


3481
ACCAACCGGT TAATCCTATA GCACTGATCT CCATTAGAAC GCCTCCGGTG ATCAACAAAA


 


3541
CCGGTTTAAT ATAGCTAGAA AAGATTTTTA GCAGTTTCAT AGTATTCATT TGTACGTTCT


 


3601
TGCTCCTTTC TCTTAGTGAA GTAATCATAA CCAGCCGCCC AAGCATCCAT AAGAGCAGCA


 


3661
AGGCAATCTA TTTTATTTTG CTTTCTAGTC TTGTCTAATA ACGCTTGACC ATTAAAGTTA


 


3721
TATTTCAATA TTGCATTTCC TACGTTGTAA GCTAACATCT GATTGTTAGT GTGAAGAATG


 


3781
TTACCAGCAA TTAATTCTTC CTTAAAAGTC CCAATTGGTA CCGATAAATT CTTATAGGAT


 


3841
TGTGTTAATT GAAGTTGTGG TAGTCCTGCC CGTTCAAAAC GATTAACAAG ATCAGCCCAT


 


3901
TTATAAGGGT CAAATACTAA GAATTTAACA TTTAGATTAT ACTTACCGAT TAGGTCCATC


 


3961
ACGAAATCAT ATACGGAATC ATAATCAATC AAGCCACTTT CTAATGAACT GATTGAACAT


 


4021
TCTCCACGTT TAGCAGCCAT TTCATAATCA ATTCCATCAC GTTTTGATTT AGTAACAATT


 


4081
CCACCATAAC GAGATACAAA GGCGTGGGAA TCAGCGTAAA ACTTTCCATC TTCTAACGGA


 


4141
ACTAACCAAC TAATCGCCGT TAAATCTCGG CTTTTTGATA GATCAGCACC AATATAAACA


 


4201
TCTCTATTAC TAATATCTGG AGCTTTTTCA ATCAATCCAT GTTCCCAATC TTGCGTACTA


 


4261
ATATATGAAT TTTCATTCTG CTGTACAAAC ATATTCATAT TCTTTACAAG TACGTTATTA


 


4321
ATATTATTTT GTTTTGTGGC TACATCTAAA TCATTTTGAA TGTGTTCCCT CATTGTCTTA


 


4381
GCAATCTTAG GATTACTCAT CAAAGGATTA GCTTTAATCC ACATTGTGGG GTCGAATATT


 


4441
TCTTCTTTTT TATCAAGACA ATACATAACC GCAAAGTAAC GATCATTCTT AATTTTGCCG


 


4501
TTGAGTATAT CGGCTGCATA GTCCCATTCA TTTTTGAAGG GTGAATTAAG ATTAAAACCA


 


4561
GCAGTAGAAA TAATCGCTAG AATACCATTG TCTTGTTGAG CCATACCTGA ATTCAAGGCG


 


4621
TTTAGAATAG TGTTATCTTT AGCTTGGTGA TATTCATCGA CTACGCCCAG TGTCGGGTTA


 


4681
TAACCGTCCG TACTGTGGGT ATCAGTAGAT AGTGGAATTG CGAAGCTACC TGTCTTTTTG


 


4741
TCAGTCACCC GTTGTTTATT GATTTTCACT CGTTGCCTTA AATACTTTGA ATTACGCCGT


 


4801
AAGTTTTCTA GTTGGTCACT CATAATATCA AACGCAAGGT GTGCCTGCTG GGAACTGTTA


 


4861
GCAGTAAATA GTATTTGTCG ATTACGGGCT GGTTGCTTTT CCATAAGAAG AGACACAACG


 


4921
CCAATCGCTG CCAGTAAGAA GCTCTTTCCA TTTTTACGGG CTTCACTTAA TAGAATCCGA


 


4981
TTGAAGCGCA ATGAATTATC GTCTACCTGT CGCCAACCAT AGATATTAGA TAAGATAAAC


 


5041
ACTTCAAATG GCTCTAGCTT CAATGGTTCA CCGTTTGTTT GTGGTAAGCT CTCAACAAAT


 


5101
TTAATAACCT TACCAGCATA CGTTTCAGAA TAATAATAAG GAAAGTCATC GGTCTTTTGC


 


5161
TTTTTCATGT CGTCTAAGAA GCGCTGGCAA GCATGAATAA CTAACTTACC GGCGATTAAC


 


5221
TCACCTTTAA CAACTGAATT AGCATACTGA ATAGCTCTAT CCATTATGAC AACATCTCTT


 


5281
CAAACTCATC AGTAGGAGCT TCTTCACTCA TTGCCTTGTG GACTTCTAGG CGTACCCGTG


 


5341
ATTGTGGGCT AAGTGCCATT TGACTATCAA TTGACTTCAT TTCTTTAATA GCCTTATCAC


 


5401
GTGCCATGTA GTAAGGTGAC TGCTGTAATC CTCGTTTTGT ATCAATAACT ACCCCAGTAC


 


5461
GATCAATCTC TTTAGTACAT TTGATAATTG TTGAATACAA AGTACAGTGA GTAGCGATTA


 


5521
AACTAACATC TAGTTCGCTA ATAGGGGCAC TCTTCTTTAA GAGTGGGACA ATTCTACGCC


 


5581
ATTCCTTTTT TGCTACATCA TCAAGCCATT TAGGGGCGTG AGATGTTAAG TTTGGGTATT


 


5641
CTGCCAATGC TTTTTCAGCG TCTTCACGTT GTGCTTTTTC TGCCTTAGTA ATATGACCTT


 


5701
CTGGACGGGT CTTCATTGGT TGAGCCAACT ATTTAACCTC CTTCTATTTA TTTGGTGACA


 


5761
CCATGTCACT ATTTGTTTAA AAAAATAAAA AAAACATCAT TTCATATTCA ATTATCGTAT


 


5821
TATTTAACCC TTATTATACC ATATTTGAAT AGTTTGTTAA GTTCAATGCC TAAAAGTTAT


 


5881
AATAAAAAAT CCTTGACATT TGAGGCTCTA AATTATTTAA TAAAGTTGAA AAGAAAACCA


 


5941
TACAAAATAA AAAGAAACGT CTTGAAATCT TCGGATTTTA AGGCGTTTTT TTGTAAAGGG


 


6001
AGAATTTTTT TAAAAGAAAC CCCCACCATT ACGGCTCCTA CCGGTCTATA CCAATAGCGG


 


6061
GGGTATTAAA ATTATTTCAG CTATTCATAT TTATTTTCCT TTCTCGTTGA TGATCTTTTC


 


6121
TTGTCTTTTC TCGGTGACAA CGATAGCAAA GTGATTGTAG ATTAGTTGAA TCCAATCTTT


 


6181
TACTCCAATC TTCACGTATA GGTACAATGT GGTCAACTAA ATCAGCTTTT TTTACTATTC


 


6241
CCTTTCGTAA ACACATAACA CAAACTGGGC TTTTCTGGAG CGTTTGACGG CTTAATTTAA


 


6301
CCCACTTACT CGTTTTGTAG AATCTATTAT AGTCATGAGG AGCTTTGGCG CTTCTTTCCA


 


6361
TTCGCTTTTT GTATACTTTT CTATTGTATT CATGTTTCTG TTCTTTGGTC ATCTTTTTAT


 


6421
AGTTATCTCG TTGCCAATGT TTACGCTTGT CTTCATATTC TTTTTGTAGA TATTGATGTT


 


6481
TTTTACAAAA ATATTGACCT ACTGGAATCA CCTCTCGACA ACCAGCAGCA CTACAAAACC


 


6541
TTACTACACT CATTTGATAA CCTCCTATGA AAAAAGGGCA TCGCATAACA CGACACCCTT


 


6601
AGCACTTACT TATTAATTAA TTCCATTGCC TTACTATATG CTTCAAGCCT GCTTATGTGC


 


6661
CAGCTTTTAA CATCGTTAAC CGTTCTCTTA ATGCAAGCTT CAAGCTCCAT TGTATCGGCG


 


6721
CTGTAAGGGT CACACTCGCT TAGTCTATAC CTTGCCATCA TCACCCAGTC TTGGCGACTT


 


6781
TCCTCTGCGT CCTTGCATAC TGCCACTTGT GCAATACACT TTACTAGCTC TTGGTTGTCT


 


6841
GGTAGTCCTA GATCAATCGC TAATGGTAAT GGACTGCCAT TTACTGAGAC TAATTTTAAA


 


6901
CTATAAAGCG TAGTATACAT ATGATAATTA GAAGGATCAT CACCGCTTGC CTCAATCATT


 


6961
ACATTAATAA AGTTGTTATA CTCTGGGTGC GTGTTGACTG TTAATTTACT CATCCCCATT


 


7021
TCACCTCTTA TTTATGACTA TCAAGCTTGC GTAAGGTTAC AATATCATAC GCCAATAACT


 


7081
CATCATCAAT TGACCAACCA GCAACTTTAT AGATACTGCC ATTGTATTGT GCTTCTGGAT


 


7141
ATTTCTTCAT GTCCGTTAAA TCTTTCTTGT GACGAACTAC CAATATAACA TCATCAGTAA


 


7201
TTGAACTACC TGCCTCACTA ATAGCTTGAC TAGCACTGAT TGAATAGTTA CCACAATGTA


 


7261
CTGTCACATC TCCTACAAAA CCTGTCATGA TCATACCAGT ATTCGGGTTA CGTTTCCCCG


 


7321
TACCACCTTC ATGACCGAAT GTGACCTTAT GCTTTAATCG TGTAATTGCG TATTTATTAC


 


7381
TTCGCATTTA ATTATTCCTT TCTGCCTGTA CTTGCAGATA CAAGCAATAG TAGAACGTTT


 


7441
GCAGCCGTTC TAGTGAAATC TACTGTTACT TTATTTTTAA CTCCTTAATA GGCTTTTCAC


 


7501
CTATCCGGTG GTATGTGGTC ACTGCCTCAA AATAAAAGTA CGGTAAAAGC AGTTTTAATT


 


7561
GTTAATTTTC ATCTCTTCAA TTGAAAATAG TTTTTACTTT TATCGGTTAC CCCGTTTCAA


 


7621
GTACCCACAA TCGCCTTTAT ATCAACATTT ATATGGTAAC CGATAGGATA TAAAGGTAAC


 


7681
CGATACATTG CAGTTACTTA ATATAGTAAC CCGAAACTAT CGGTTACCTC AAAATGCTTT


 


7741
CGGTTACCTT TAAATAGCTT TATATCAAGC TTTATAAGAA CTTCTGCAAA GGTAACCGAT


 


7801
AAATATAAAA CTATAACTAA TTTTTAAGAG CAATTCCTTT ATAACCTTTA ACATTTTTAC


 


7861
CGTTAATCTT AACTGTAGTG TGTTTGTATC CTCGTGCTAT TAGCTCTTTC GCAAACTTAC


 


7921
GATTACTAAG ACAAGGAACA GCAGTATTTT TACAATACTC TTTGTAATTA TCATATAGAT


 


7981
ATACTGATTT TTCTTTATCT TTCTCATTTG TAGTACAGCA ATCATTAATC CAATTACCGA


 


8041
TATTGTCATT AGCATTTACC CAGCTTCGCT TTTGCTGTTC CATGTATTCA GTCACTGGAA


 


8101
ACTTACCAGA ATCAAGTGCT TTTTTATAAG ATTTTAAGCA TTCATAGGCA AATGCTGGCA


 


8161
TTTCTCTTTT AAAGTCTTGC TCTTTGAATT GCTCTTTAAA ATGTTTAATC TTATGGAACG


 


8221
TTACAATAAT TGGTCGTCTT TTAAATCCGT CTGTAAAATC ATTGAAGGCT GGTAAATCAT


 


8281
TAGCGCTAAA AATCAATTTA GCTTCATTTT CAAGTTTAAA TGGATCTTTA TATTTAAATT


 


8341
GAGCTGTTGT AGTGTCGTCA CCAGTTAATG TTTTAATAAT ATTAGTCTGT TCCATAAATT


 


8401
TGGGGCTTAT ATCAGCAAAA ACATTAGCAG CTTTATGATA CAGCTGAGAA GTAGCAAATT


 


8461
TAGCTTCTTT TTGGTCGGAT AATGCTTCTA AAGAAACGTT TGAAACGTTA GGCTTCCCTA


 


8521
TTAACTCCAT TAGTTTATTT AGAAATGTAG ATTTACCATC CCCACCCAAC CCGTATAGAA


 


8581
TCATATAAAT TTGAAAGTCC TTAAAACTAC CTGCTAAAGC AAAACCAATA AAGGCTTTTA


 


8641
CTGTTTCAAT TGCTGTTAGG TCATATTCAT CTTGTAGTTT TCCAGTATCA TCACGGATTT


 


8701
CTTTAGTTGA GGGTACTAAT GATTGTTTTA ACCATTTATT CCACGTTGTA GCTTTTCCTT


 


8761
TTGTTTCTAA TTCATATGGT CTATTTTGTA AAAGATAGTT CTCTGGTGAA TTATCTTCTA


 


8821
AAGTGTCCGT CATCAAGTTG TAAGTGTGTT TACCAAACAC TGCTTTATAA GGTGCTGGTT


 


8881
GATCAAATGG ATTGTCTTCT ACAATTTCTT GTGAATCTAA GGCAATTAAT TTAGAAACCG


 


8941
ATGCTAATAA GTTCGTTCTC CATATTCCAT GAGGTACAAT TTCATTTTTG ATTGATTTTT


 


9001
CAATGACTGA ATCAGCGTAA TTTTTCCATT GTCTACTTTT AGGATCGAAC CAATAGCCGT


 


9061
GTTCTATAAA TTGGTTGGAT GTAAAGTGAT ACTTTTTTTC TAAATATTTA GCCAACTTAA


 


9121
TTTCATCAAC CAGTTTATTT CCTTTATCAC TTACAGTTAA CCATTCTGGA AAACTATTGT


 


9181
TATTTACTAA GTCATCAATA TTAGCCATAT TTTTCCTCTC TTTACATGGT AATTTAGCGT


 


9241
TTGTATTTAA GACTGATAAT CTCTACTTCG CCTTTTGTTT TGATATATTC AAAGCTTTTC


 


9301
CCGTCTAAAC TTGTTTCTTC GTATATATTG TCCCAATCGA CTATTGATTG ATCTTCGTTT


 


9361
TGACTAGCAT TATTCCAATA CTTAAAGAGT GGTGTACGGT GCAGTATTCG GTTAACTTGT


 


9421
TTAACGTCAT CCCCTGTGTA GTAGAGAAGA TAAAAACACA TTGCACCAAT AGCAGGTTTA


 


9481
ACGCTATGAT ATTTATTTAG GTAGTAGGAA ATTATTTCCT CATTATTTTT ATAAAGAGCA


 


9541
ATAAATTTTC TACTACTTTT ACTACTAACA GCTCGTTCTA TAACCTTGTA ATCATCGTTA


 


9601
TAGTATTCAA GTGCTTTCTG AAACTGTTTC TTTGCGTACT CATTAAGGGG ATGTGACGTT


 


9661
GAATTAATAA TTTTTTCCAT CTTGTCACTA TCTAATAAAG GATTAACGGA TAGTTTTTCT


 


9721
TCAAAACTTA ACACTATTAA TCACGTCCCA CCTATTGCAA TTGATCATGA ATTTGATCTA


 


9781
ACTTATCCAA CAAAATACTA GATAGTGCTA ACAATGCTTT CTTACGTGAT ACTACTGTTT


 


9841
GAGCAACCGT AGACACTTGT TCATCACTCA TTGCTGCATA ATTGTCTACT CGTACAATTG


 


9901
GAGCAAAAAC ATCATTTAAC TGATCATACA GTGCTTGGGC TATTCCGGTT GTTTCTTCAA


 


9961
CATTGGAATC TGCTTCCCAC AACTGATTTT CAAACGCTAG TTCTTGGTTC TTATCTTTTT


 


10021
CCATTGTTTA AACCCTCCAC TCATGTTAGA ATGAAAGAGA ATTAAATATT AGTAATTCGT


 


10081
CCAACATCCC CCGTAAAGAT TGTTGGCTTT TTTATTAGCT AAATTTAATT CTCTAAGTTT


 


10141
TATTTAGTCG TTGTTAGTGT CTGAATTGGC GTTCTCACTA TCAACGGCTT TTTTATTACC


 


10201
ATCCATAAAG TCGTTTTCAA AATCAGTTAA AGTTTTATAT TGAATTGCTA AATCTTTAAT


 


10261
TTCTTTAGCG ACCTTAATAA TTTCATCTGC TGAGGAAATC GGATCATCTG CAATTATTTC


 


10321
TCCATTTTCT TTAGTGATAG TACCTTCAAA ACAATCTTCA GTAATTGAAT TTATATAACT


 


10381
GCCAGCTTTA ACAATCCTGT CTTCTATATC AATAATTCTT TTACTTAATA TTTCTTATTC


 


10441
AAATTTTTTA GTGTATAAAC TCATTTCTTA TTACTCCTTA AAATAAGTTA CTAATTGCAA


 


10501
CAACAATACC CAGCAATGGT AAAAACGTTG CTAGATAAGC CCAAAGGCGT TGGTCTGGTG


 


10561
AAGTCCACCA GTAAGCTAAA ATCTTACCAT CAATTTTTTG TAACATATGT CTCACCTCCT


 


10621
CCTATTTATT TCTCTACAAT CACTTGCCAG CCAGCTGTGC GATTATTACT TAGAATTTTC


 


10681
TTTTACCCAA CGGTCAACAT CTTCACGTTT ATATACATAT TTTCCTCCAA CCTTTGATGA


 


10741
TGGAAAATTT CGGGTCTTAA TCCAACGATC AATAGTTGTT CGAGAAACAC TTAGATAATC


 


10801
CGCAGTTTGG CCCTTATTCA TAAACTCTGG TTCGGTTTTA ACAGTCTTAG CATCTTGTAA


 


10861
CTTATTGTTC TCTAGCTTTT TAGCCACAGC TTCAGCTAAC TGATCAATGA ACTGATCTAA


 


10921
AATAGCTGTA CTCATTATTA TCACCTCTTT ATTTCATATC GTAATTACAA TATATAACAT


 


10981
AATGAAATAT TATGCAACAA CAAAATAAAA TATATTGTGT TTTGTATTGT TATTTTCGGT


 


11041
TATGTATAAT ATATTAACTA TACAAGGAGG AATTAATGTG AAAGAAATTT CATCTGACCA


 


11101
GAAAATAGGT TCTAACATTG CACTAATTCG TAAAGCTAAA AAGATGACTC AAAAAGACTT


 


11161
AGCAAGAAAA ATTGGAATAT CACAACCAGC CTTGGGGAAT TATGAACGTG GCCAGCGGAT


 


11221
TATACCTGCT TCGATTTTTC ACAGTTTACC TCAAATATTA GATGTTCCTC TAGATTTTCT


 


11281
AGTCTATGGA ATTCCCAACA ATATTACCTT TCATGATTTA GCAACGCTCG ATTCCGTATC


 


11341
TCTCAAAAAG AAAGCTGCTC ATATTGAAAA TGATAGAAGT ATTATTGAAC GTGGGTTAGG


 


11401
GCGTGACGAC ATAAAAAGAC ATTTATTAGC TGCTGTTTTG CTTGATAATT ATGGTTATCT


 


11461
TGTGTACCCA GATGAATCTA GCAATGACAA AGAACATGAA CAACTTAAAA AATTTGTTTG


 


11521
GAATACATTA GATGGATTAC TTACCGGCGT ACCAATTACC AATGAAATAA ATTTTAAACA


 


11581
AATTAATGAA CAATTTGAAT TACTTGCAAA TACGTATTTA AGCCTAATTA TTGACGAAGA


 


11641
GGCAAATCGC TATCCTGCTT TCCGCAAGGA TTTACCACCC TTCAAAGAAA AACTTACACA


 


11701
GTTTTTAAAT GATAACGGAT ATTTTAAAAT TTTAAAAAAA TAATTTTAAA CCTTTCTCTA


 


11761
CAATCTCTTG CCAGCCAGCT GTGCGATTGT AGGAGGAAAG TAAAAATGTC AGTATACAAG


 


11821
GATAAAAGAA ATGGTACGTG GTATGTTAAA TACCGCATTA ATGGTAAATC AACCAGTAAG


 


11881
CGAGGTTTCA AATTAAAATC GGACGCTGTA GCGTGGGAAC AAGAAAAAGC TGTGGAGCTT


 


11941
AAAAAATATG GTCAAATGAA TAACCATAAC ATGACCGTTG AAGAGCTATC AAAAAAATGG


 


12001
TTGCCAGTAT ATGAACAGAC TGGGATAGAA AATTCTACTA TTCATAAAAC AAAACAAATC


 


12061
ATTAAAAATC ATATTCTACC CCGCATTGGA AACTATAAAG TTGCTGATCT TAGCATTGAG


 


12121
TTATTAACTG AAGTTGCTGC TAAATGGAAA GAAGAATTAG TAAAGAGTGA CCCTTTTAAC


 


12181
TATACTAAGC GAATGTTAGA CTACGCAGTA CAATTAAGAG CGATCCCAAC TAATCCAATG


 


12241
AATTCTGTAG TAAAGCCTCG TAAAAAACAT GACAAAACAT TTACTAACAA TAATTTCTTT


 


12301
AATGAAAAGC AACTTCACAA TTTTATCAAG TGTATAAAAA AAGACTACGA AGAAAAAAAT


 


12361
CCTCGTGCTT TCATGGTTCT ATGGTTAGCC CTTTTTACTG GCATGAGAAA ACAAGAACTT


 


12421
CAAGCCTTGA CGTGGCAAGA TGTTAAATTT ACAAAAAGCG GTGGAATAAT CCATATTAAC


 


12481
AAGGCTATTA AGAACGCAAA ACATCCCTAT CTTGGTGGCC CTAAAACCGC TAACTCTTAT


 


12541
CGCTGGATTT CAATAGATAA AAAAACTGCT TCCTATCTAA AAAGATGGAA ACAGCAACAG


 


12601
ATAGATATTT TAACTAAACT AGGTTTTAAT CCACACCAAA AAGAGCAATT GATTTTCTCT


 


12661
ACCTACACTA AAAATAAAAT TGTTTTAGGT GCAGAATTGG ACAAGCCGTT AAATAAAGTA


 


12721
ATTGTTAGAA ATAATCTAAA GAAAGTTACA TTCCATGGAT TACGTCATAC CCATGCTACA


 


12781
CATTTGGCTT CTATTGGTAC CCAGCCAAAA TTAATAGCAG ATAGATTAGG CGATACAATG


 


12841
GAAACTGTTC TTGAAGTTTA TATTAATGCA GATACTGAAC CAGAACAAGG AATCGCCGAC


 


12901
AAGTTCGCCG ATAGTTTAGT ATAA.






Bacteriophage loci 6 has a sequence of:









(SEQ ID NO: 13)










1
TTAAAATTGA CTAAGTGCCT GGACGATTTT GTCATCAGTT TTACTTTTAT ATTCATCAAT



 


61
CAAGTAGGCG TATGTGTTCA TTGTGGTTGT GATATCATTA TGTCCAAGTC TCTTGCTAAT


 


121
TGCATAAATA TCTATTCCGT TACTTAACAG TAAAGCAACG TGACTATGAC GTAAACTATG


 


181
GAAATGAAAA TTCTTCTTTG ATAATTTTGT ATCACTCATA ATTTGTCGAA GCGTTTTATT


 


241
TAATGCTGTA CTAGTAGGGA TAGTATTAAA TTGATCCATA AAAACAAGGT TACTTGATGA


 


301
ATTATTGCGC AATTGTTTAA GTAGTAATAG TAGTTTTTCA TTAACTTTTA TTTTACGTGT


 


361
AGATGATTCA TTCTTGGTGG GTTTAAAGCG ATGTGTATTA AAATGTTCGT CTGTTTCGTC


 


421
TCTTCCTTTT ACCTCTCTCC ATGCTTTATC AATATTAATG GTATGATGAA TGAAATCAAT


 


481
ATCATTCCAG GTAAGAGCTT GAATTTCTTC TTTTCTCATT CCAGTATATA TCGCAGTGAC


 


541
TATCATATAA CGGCTTGTAT ATCTTCTATT AGTAATTCCA TTGATTGTAG TAGTTAATAA


 


601
TTTCCTTATT TCAGTTACAT TAGGGTATTC TACCTTCATT GTTTTGCTAG TATTGGCAGC


 


661
TAAAGTAACA CCTTTAGTAA AATCTTTAAG CAGATAATCA TCAAGGATAG CAGATTGTAC


 


721
GCAAGACCGT ATTATTGAAT TAAGCTTCTT TACACTTGCT ATAGCATGAG TTGCTCCATA


 


781
CTTGTTAATA AATTCTTGAT ATTTAGAACG GTTAATTTCT TTAATAGAAG CTTCTTTGAA


 


841
ATAATCATTA ATTAGATTAC CGATTATAAT ATAACGATTC AGAGTTACAC TTGATATTTT


 


901
AGGTTGCTTA TAAGTATTAA CCCATTCTTC GTAATAATCA CTAAAAGCAA TTTTTTTCTC


 


961
AATGTGTATT CCGCGATTAA GATTAGTTTC CATTTCAGCT GCCCACTTTT TAGCTAGTGC


 


1021
TTTTGTTGCA AAACCAGCTT TGGACTTTGA AAAACGTTTT CCTTCAGTAT CTTTCCACTG


 


1081
AATTCTTACA GACCATTTTC CGCTTCTTTT ATATATTTGA GCCATACTTG TCCCTCCATT


 


1141
ATTAAATTGT GTATAATGAA AGGGTTGATA GAATTTGTTT GGACGCAATT AATATCAACC


 


1201
CTTGGTCCAC TGACAGTTGC CGCTGTCGGT GGGCTTTTTT AGTTTAAATA GCTTTTTACG


 


1261
TCATCAATTT AGTTGGACGA GAAATTTAGC TTTTGTTTAG CTTCTTTAAT TGATTAATAA


 


1321
TTTCTTGATT TTGCTTAATT GTTATCCAAT TTTGCTGTTT GATTATTCTT AATTCATGAA


 


1381
AAAGGGCTTT TGTATCATTA CTTGTAAAAG TAGGATAGGC ACCTGTTAAT TTTTCGGCAA


 


1441
CTGCTATATC TTTTACAGAT TCGAGAATTT CGTCATCTAA ACTTTCTAAA CGATTTTCTT


 


1501
TAATATATTT ATCTTTGTCT TTAGCAAATC CATCTTTACC ATTGAAAATA CTCATTTTTA


 


1561
GCTTTCTCCT ATCTTTTTAA TTTTTACATT GTAAAGAGCG TTATTTAAAT CATTACAGTT


 


1621
CCTAAGATAG GAAAAAGCGT TTTTTAAAAG TTTATTCATC CTTTGACTCC TTATATTCCT


 


1681
AAAATTTGTT TCTTTTTAGC ATCAAATTCT TCTTTGTTAA TGATTCCATC ATCAAGTAGT


 


1741
TGTTTTAATT CTCTTAATTG CGAAAGGTCA TTACTAGAAT TGGATTGTGC TGATGTCTGC


 


1801
TTAAAATCTG CAGCTGCCTG TTTAATAGTC TCAGCCATCT TTTTAGCAGG GTAGGGTTGC


 


1861
ATATTTTCTA TTTGAGTTGT AATAGCTCCG TTGGTTACTG CAATAGATCC CAACATTAAA


 


1921
CCTTTAGAAT ATGAAACGCC ATTAATCATA TCAAGGGGAA TGTCAGTTGA TTTACTACCA


 


1981
TAAATCAGGC CGTGGTCAAG AAAAATTACG CGCTTATTAG TACAAACCGC AAGAATGGAA


 


2041
TTAGTTTCAA CGAAAGCATT AGCAGCATAA AGTATTTTTT CTCCACCATC GACATCTATT


 


2101
ATGTCTGGCA AGGCTTTGAT TTCCTTTTTA GTACCGAATA GATTTTCTAC ATTGGCATCC


 


2161
TTTAACTGTT GCTTAATAAC ATCCAATTTT TCTTGTTCGC CTTTGTTGAT TTCGTCAATT


 


2221
GCTGCTTGTA AAGTATCAGC TTTTTTACCA GCGCTATCAG CAATTGAATC AAGTTTCTCA


 


2281
GTTACTTTAG ATAGCTTTGA TTTATACAAA TCAGGATGTT CTTTGTAAGG TACGGTAGAA


 


2341
TGAATCCCTA ATAAACTTTC AATTTCACTA GCGGTATGGG AACTAATGAA ATTGCTAAGT


 


2401
TTAATTGGAA CACGGTAAGT ATTTGCTGGA CGTTTGGGAT CAAGTCTGTT GGCATCTTTG


 


2461
CCACAAATTA AATATCCATC TTTGGTAATG CAAGAAGCCT TTAACATTCC CAGGGCATCA


 


2521
TTACATATGA AGCATCTCTT TTTACTCATT TGGATCACCT TATTCCATTG TCACTGAATA


 


2581
TTTAACTAAT TTACCAACAA TTCGACCTGG ATGCTCTTCA TCAAGAATTA TTGGATCATA


 


2641
GTTTTTATTA TCTGGCATTA GCATAACTAA ATTGCCTTGA TGCTTAATAC GTTTTAAAGT


 


2701
AGCTTCGTTA TCATCGTCAA CAAGAACTGC AGCTATCTCA CCATCTTCAA CTTCAGGTTG


 


2761
CTCTCTAATA ATGGCTAATG CGCCATTAGG AATGGTAGGC TCCATGCTAT CACCTTTACA


 


2821
ACGAAGACCA AAAAGTGTTC CACTAGGAAC TGGTTTTTCA AAGATTTCAT CAGTATAACC


 


2881
TTCGATATTT TCTTCAGCAG TAATCGGATC GCCACAAGCA ATTTCACCAA TGATTGGAAT


 


2941
TGAAACAACT TCTAATCCTT CTTTAGGATA AATAATATTA GAAGGTTTCT TTTCTCTGGT


 


3001
AGGGAAAAAG TCATCAACAT TTACATTAAA AATTTCAGCT AATTTAAATA AAACATCTTG


 


3061
ATTTGCCTTT CTAGATCCTA ACTCATATCT AGTCACAGTT GCTCTTGTCG TATCAAGTTT


 


3121
GTCTGCAAGT TGTTCTACTG AAAGGCCACG TTGTTCACGA AAATTTCTGA TTTTTTGACC


 


3181
AATGAATTTA CTTAGTTCCA TTTTATGTAC CTCTCTTATG TAACTAATTA TATAACTTGC


 


3241
GTTACCGAAA TGGAATATTT TTTATCTTTT TCGAAAAAAA TAGTTGCATT TGTTCCAGAA


 


3301
TGGTTTTAAT ATATAATTGT TCCAAATTGG AACGGAAAGG AGAGATAATA TGCAGTCAAG


 


3361
ATTATATGAT TTACGAAAAC ATGTTAAAGG AATGACTCAA CAACAAATGG CTGATTATCT


 


3421
AAATATTAGT GTCAAAGCTT ATCGTGATAA GGAAAATGGT AAAAATCAAT TTACTCAAGA


 


3481
TGAAATGTTT GCTATTAGTA AATTATTTGA TCTTAATATT GACACTATTT TTTTGCCTCG


 


3541
TAAGTTCCAT ATTGGAACAA AGGGCTAAGA GAGGTGACAC AAAATGCAAG CATCGCTAGA


 


3601
TGAACAAGAT TACCAAGTAA TTACTAATGA GGTTCTCAGG CGTATTAAGG AATGTTACAA


 


3661
CTTAGTGCCT AAACAAGATG TCCAAACTGA CAAATGGGTC GGTATCAAGG AATTCACGAG


 


3721
TAAGTTGCCC GTTATAAAGG ACAAGGAATG GGTCAGAATG TTTCTCCTTA CACTTCCAGT


 


3781
CTTTAAGAAC TGGGTTATCA ATTTGAATGC AGGTCAAGGT CATCGAACTA AGGTAAATGT


 


3841
GACCAAGGCA TTGCCTTGGA TTATGTCTCA TCAAGCTGAT ATTGATTGGA ACCAGTCATT


 


3901
GCCACGTTAG GAGGTGAACA GATGGCACAA CCGATATTTG TAAAAGGAAT TTTTGGAAGA


 


3961
CGAATTATAG TTTCTCAATT TGAACAAACG TCTGATTTTA TTTTGCAAAA GAAAAACGCT


 


4021
GACCTAATAA AAGAAGCCAG CGGATTTTCA AAAAACAAGA AATTGTTTAC TCGTTATCAA


 


4081
GGTAAAGACT TTCAAATTAT GGAGCAATTA AAGTGGCAAC CCAGTGTTCT TTTTGATGAT


 


4141
GCCAGTAAGA ACCGTAGTAG CCACAGATTT AAGGACACCA ATGGAAACTG ACTTTAGAGT


 


4201
GTTAGTAGCT TTCTTTGTTT GTTTCCAAAC CTCATCATCT CGAATGGAAT CAAGTAGTTC


 


4261
ATGCCCAGAC CAAGTGATGT CATAGACAGT TAATTGGCGA ACCATTTTAT TAATAAATGG


 


4321
TTTGGCACTA ATGAAACCGC CTGAGTAGAG CTGATAAAGG GTATAAACAA TATCATTTTT


 


4381
ATCAAAGGTC TCATTAGCAA TAACATTTGC AATTTGAGAA TCTGTAAAAC CTTGGTAAAG


 


4441
CATTTGCAAT TGCTGTTTTT CTAATGATAA AAGAACATAC CTAACACAGT CTTGCTTCAA


 


4501
TTTCACAATT TTCACCACCT TTATTTTGAA TTAATCAAAG TATAGCAGAA AGGGAGAGCA


 


4561
GTTAAATGGC GTTGGTAATT GGATTACCAA TATTAATTAT TGCAATGTGT TTGTTATATG


 


4621
CATTGGTGTA CAGCTTATTG TATGAGCGGA ATAAACCATT GCTATTGAAA GAAAAGTATC


 


4681
GGAAGAAACA TTGAAAGGAG GTGATATGAA CTTTGGTAAA TTTATTTCTT TTGTTGTTTT


 


4741
TTATCTTTGT TATAATGCCG CTTTTGTATC AATTTTTGAA TTACATCAGA AGGCAAATTG


 


4801
AAAATTTCTT TTGGAAAAAA AGCCATAAAA ATAAGAACGA ATGAAATGAA AGTAGCTGTA


 


4861
AATTCAAATT GTTCTTTGGT CTGAGTTATT AATAGCCCAG TAATAGTAGA AAGAATAATT


 


4921
GCTATTTTTG AAAGATCATC CAAGTATTTT TCGCTTATTT GAAATCCTAA AGTTTCTGTA


 


4981
AGCCAGCCAA GGATAAACAA TGGAAAAGAA AAGAAGCCAG TTAGAAAAAG AATAGTAATT


 


5041
AAGCAATCAA ATATTCTGAG TTTTTGGAAT TCAAAAATTA CAGCATCTGG ATTAGCAGCA


 


5101
TATGTAATAT GTAGAGCTAA ATAAGTAAAA CAGATGAGGC AGAAGATACA AACCATAAAG


 


5161
TCGGTTAGTT TCTTACTATA GTAATTAAAT TTGGTTAAAA ACTTAAAAAT TTTACGCATA


 


5221
ATGATTTTCC TTTTGTACTG AGTTAATCAA AGTATAGCAA ATGAAAGGAT GGTAGTTTAA


 


5281
TGAGTAAATT AATTGCGCTA ATCGTTGGTG CTTGGATCAT GTATTGTTCA ATGATTGGCT


 


5341
CTTATGATGG AGCGATGGCG ATCCTAGCAG TTTATCTTCT GCTAGTGGTT CTTGATCCAC


 


5401
TAAATAAAAA AGGTACGACT GCCGCAAACA GTCGTACCGA TAAATAAATA ATCTACGAGG


 


5461
TAATTATATA TGAATGAGTT TAATTTATCA AAATTAAATG CCAGGGTTGG CGATAATTGT


 


5521
GTATTTGTAT CGAATTTGGC AGTTCGCTAT CAAAGCGCTG CTACTCCTGA AGAGCGAATG


 


5581
GCAATGGCCA TCAAAATGGA GAATGCTGCT ACGATGTTAC GCATTTCAGC TGAGCGTTTG


 


5641
GCCACTGAAA CTAAGAATGT TTACGGAGGT AAGAACAATG ACTAACGAAG AAAAGATTAA


 


5701
GGCAATTAAA CAAATTCTTG GACCAGAATA CGAGGAGGTA GCCATTTTCG CTGCTAAAAA


 


5761
ATCAGAAATT CGTGACGAAC GTACTAATTC ATTAGTTGAT GCGGATCCCG GAACGGTAGC


 


5821
AGCCATGATT ATGAATTGGT TACATACTAA TCCGATAGCA GCTTCAATTG TCAAAGCAAC


 


5881
TATTGATACT TTGGAAACTG ATCCAACTGC TGATTTATTT GCACAGATTT TTCTAGGAGG


 


5941
TAAGAACTAA TGAATTTATT TGAATTAAAC GACAACTACA AAACACTTGC TAGTCGGGAT


 


6001
GACTTAGATC CAACTATTCT AAAGGATACA TTGGAATCAA TTAAGGATGA TCGGAAGACC


 


6061
AAATTGGACA ATCTTGCTTC ATGGGCAGAT CATTTGAAGT CAGAAATTGA TTTTATGACC


 


6121
GACAAAAAGA AGTCATGGGA AGAAGAAATT ACTTACCGAA AAAATAAACT TACTTGGATC


 


6181
AAGAAATATA TTACTGAAGT TCTTGATGAT GCCGGTATTA AGAAAATAAC TACTGAAAAT


 


6241
CACTTACTTA GTGCCCGGAA CTTTAAGGCC TCAACCATTA TTGATAGTGA TAAGAAGCTT


 


6301
CCGGATAAGT TCAAAATTAC TGAGACGACT ACTAAGCCAG ATAAGCAGGC CATTTACCAA


 


6361
GCACTCAAAG CTGGAGAAGA AGTACCAGGA GCACATTTAA AAGCTAACCG TAACACGGTG


 


6421
ATTAAATAAT GTTTGAACTC CGTGATTATC AGCAAGAAAC GATTGATAAC ATCATGAATT


 


6481
CTATAAGTGC TGGTCACCGT TCTATCATGG TTCAACAGCC GCCACGAACG GGGAAGACAG


 


6541
TTATTATGGC CGAGATTGCT AGACGAGCAA CGGCAAAGGG TAACCGTATC TTGTTCGTGG


 


6601
TTCATCGGCA AGAAATTGTC CAGCAGGTTA TCAAGACTTT CAAAGCTAAT GATGTAAATA


 


6661
TGGATTTAGC TAAAATCGGC ATGGTTCAAA CGATTACCCG ACACGTTAAT AATTTGGACC


 


6721
CACCGGCGAT AATTTTTGTT GATGAGGCCC ATCATGTTCT GGCTAAATCA TATCGAAGGA


 


6781
TTCTTGATGC TTTTCCGAAA GCTTATAAGT TACTGTTTAC CGCTACTCCT TATCGGTTAG


 


6841
GTGGACAGGG TTTTACTGAT GTGGCTGATG ATTTAATTAT TGGCAAATCA GTCCCCTGGT


 


6901
TAATTGACCA TCACTTTTTA GCACCAGTTG ATTATTACGC TCCTTCTTAC ATTGATACTG


 


6961
CCAAGTTAAA AGTAAAACGA ACTGGTGAAT ATGACACTGA TTCAATCAAA AAAGCCATGA


 


7021
AGCCTAAAAT CTACGGGAAT GCGGTTAAGC ACTATTTGAA ACTTGCTACG GGAATGCAAG


 


7081
CCATTGCCTA TACCTATAAC GTTGATAGTG CAATTAAGTT AGCCAATGCA TTTAATGGCT


 


7141
ATGGGATAAC TGCAAGCGCC GTTTCCGGAA AAACGCCCAA AGAAGAACGG AATAAAATCA


 


7201
TTGAGGACTA TCGCCAAGGA AAAATTCAAA TTGTAACCAA TGCAGAATTA TTTACAGAGG


 


7261
GCCTAGATTT ACCAAATGTT GATTGTGTCA TTATGTTACG GCCAACTCAA TCATTATCGT


 


7321
TGTACTTACA ATTTGCAATG CGCTCAATGA ATCCACGGGA AGGTAAGACT GCAATAATTA


 


7381
TCGATCACGT GGGAAATGTT GAACGATTCG GGCTGCCGAC TGATGAGCGG CAATGGACAT


 


7441
TAGAAGGTAG CGGAAAAAAT AAACAACAAC CAGGAACAAC GCTTAAACCT GTATCAGTAT


 


7501
GTCCGATATG TTTTGCTTCG TTTTATCGTA CAAGTGATAT TTGCCCTTAT TGCGGGGCGG


 


7561
CATTAGGAGA AGAAAAAGAA ATTGAAGTCG TTGATGATGT TCAACTAAAA AAAGTTACTA


 


7621
AGTCACGGCT AGCGATTATT AAGAAAATTC AATCGTCAGC AATTATGAAT AATGTTGCTG


 


7681
GCAAGCGTCC AAACGAATTG AAGAATCTGA AAGAAATACA AGCCTATGCC AAATTAAAAG


 


7741
GTTACAAACC AGGCTGGGCT TACCACTACG CTAAACAGCG TGGATTTATT AAGAAGTGAG


 


7801
GTTGATATTA TGAGTATTTT GCCACCAAAT AAGCCACAGA AGGCACGGCG AGTTCCAAGA


 


7861
AATTACTTTA TCTACGGAGA TACAATGTCC GGAAAGTCAT ATCTAGCTGA ACGTTTTCCA


 


7921
AGTCCGCTAT TTCTTAACAC CGATGGTAAT AGTGAGATGA ACACCGCACC AAGTATTCAA


 


7981
TTAAAGAATG TCCGAAAGAG CGATGGAAGC TTGAAAGAGT CAGTGATTGA TCAACTAGAC


 


8041
AAGATTATTC TTGCTCTTGG TACTGAAAAT CATGGTTACA AAACAGTAGT TATTGATGTG


 


8101
ATTGATGATG TAGTAACACT AATTGAACAG GCCATCTGTT ATGACAATGG AGTAGAAACG


 


8161
CTGGGGGATG TTCCTTACGG CAAGGGATAT GCACAATTTA ATACCGTCTT TCAAGCATTT


 


8221
GTCACTGAGC TAAAAGCCTT ACCACTGAAT ACGGTTTACA TTAGCCGGTT AATGATGCTA


 


8281
ACTGATGAAT CTTCTGGCCA CACCGAAGAC CGACCATCAC TAAAACAGAA ATATTACAAC


 


8341
GTGGTTAACG GAAATTGTGA TTTAGTGATT GAAACTAAGC GCTATGGTGA CCGTTATATC


 


8401
CGGATGGTTA AAGATCGACG AATTCATTAT GTCAAAGATG ATATTACTGA TCCGGCAATC


 


8461
TTACGGGTAC TTGAACATGT AAATGGCGTC TTTGATAAGC CAAAGCAGAC TACTACAAAA


 


8521
GAGCAGAATG AAATTGTTAA CAAAATTAAA AAGCAAAATG TAAAGGAAGG TTAATGAATT


 


8581
ATGAGTTTAC GAGATGCAAT GAATAAAGCT ACTGAAGGTT TTGATCCAAA GAATGATTCA


 


8641
GTTAATAAAT TTAAGGGACT GGAAAGTGGT AAATATACCG TTGTAGTTGC AAAAGTAGAA


 


8701
AACCATGAAA CTCCTTGGAA TGCTGAACAG CTTAACTTTG AGTTAGAAGT TGTCGATGGA


 


8761
GAATCAGCCG GCCAAAAAGA ATTCTTACAA ATTGGATTAG ATGAATTAAC TTCTAAGGGT


 


8821
AATCCCAATC CAATGCTAGA AACTAATTTA CGATTGGTTT CTAAGTTAGC AGCAATTCTA


 


8881
GGTGTTGAAA TTCCCGATGA AGTTTGGGAT GACGATACTT TAATCTACGA GAACTTGGCT


 


8941
AAAGCATTTG CACCGGCAGT AGGAAAGACC ATGATTATGG ATTTGAAGGT TCGACCAAAC


 


9001
AAGAAGAACC CCCAATATCC ATACCGTAAT TATGACTTTG ATGAAGCGGA ACAGCCGGAA


 


9061
ACGCCAGAAG TTACAGATGA TGAAATGCCC TTTTAAGTAA ATATTTGAGT CAGTGAACTT


 


9121
ATAACACCGT ATGGCTGGGA GGCCATTAAG GAGGAAAAAT GAAAAATCTA GTTAATTACG


 


9181
CCTTAGCCTA TCAAGCAAAG GGTTTAAGCG TCCTCCCAAT TGCTGGCAAG CGTCCACTAA


 


9241
TTAAGTTTGC TGATCGTGAT CCACTTACCG CCGAAGAAAT AAAGACCATC TGGATAGAAC


 


9301
ATCCATATGC TCAAATTGCG TTGCGGACTG ATAAGTTCTT CGTTGTTGAT ATAGACCGCA


 


9361
ACCATGCTGA TAACATTGAT GGTTTTGAAT CAATTAAGCA ATTACCAGCG GAATATTTTC


 


9421
CGGAAACTTT AACCCAAACC ACCAAGCATG GTGGCCGACA GTTATTTTAT TTGAAACGTT


 


9481
CAGATATGCG AGTTAATCAG TTAATTGGTT ATTTACCAGG TGTTGATGTA AAGGCCCACC


 


9541
AAAATAATTA TGTTGTGGTT GCGCCATCAG AAGGTTACCA ATGGCTGAAT AAGAAGCCGA


 


9601
TTGTTACGGC TCCTAAGTCG TTAGTAGTGA ATATTAACCA AATGCGAGCC AGTAACCGGC


 


9661
GAAGTTCTCC AGATGATTTA GTTTTTAAAC CACGTGAACG CAACTCAACT ACTGACTTAC


 


9721
TTGAGACCAT CGCAAATGGA TTGGGAGATA AAGGAATAAG AAATAAAAAC TTAGCTGGAA


 


9781
TGATTGGTGC TTTATTGTTT CGGGGAGTAG AACCAAAGTC TGCTTATCAA TTAGCGATGA


 


9841
TTTGTAATGA GAATACGCCC GATCCACTAC CAGAAGAAGA AGTGAACCGG ACATTTCAAT


 


9901
CAATGCTAAG ACGTGATTTG AGAAACGGGG GTGAAATACG TGGCGGATAA TATAATTCGC


 


9961
AAACCAATTG AATTTGAATT AAATACTCAA GGCAATCCTA AAACTAATAG TTTGAAGAAT


 


10021
ATTGGTTTAA TCCTTGATGG CGATCCACTA CTGCATGGCA CCTTCAAATA TAACGAGTTT


 


10081
GCTTATTCAA TTGATGTTGT TAAGGACATT CCACAGCTAT TTATTGAAAA GGGGCAACTT


 


10141
GATGATAGCT ATTCAGCAAT TATGCTCCGT TACATTGAGG ATGAGTATGG GGTGATGTTT


 


10201
CAAGAAAAAT TGTTAAATAT GGCAATCACT GTTGAAGCAA AAAGCCACCC ATATAATCCG


 


10261
GTTAAAGAGT ATATGGAAAA GTGCTATAAG AATTGGGACC ACAAAGAACG AATCAAAGAC


 


10321
TTCCTACCAG TCTATTTAGG AGTACCCAGT GGTGAAGTAA CAACGCTGCA GACAAAATTA


 


10381
TTCTTAGTCG GAGCGGTGAT GAAAGTCTAT AAGCCGGAAA GTAAATTTGA TTGGGTGTTT


 


10441
GATTTAGTTG GTGGCCAAGG CGTTGGTAAA ACTACTTTAC TTAAAAAGTT AGCCCATGGT


 


10501
TGGTATACAG ACCAATTTAC GGACTTCAAA GACAAGGATA ATTTTGCCAA TATGCTGCGG


 


10561
GCATTGATTG TTAATGACGA TGAAATGACG GCCACAAATA ATTCTGACTT TGAAAATTTG


 


10621
AAGAAATTTA TCTCAGCTGA AGAATTAGAG TTCCGGCCAC CATATGGACG ACATACAATC


 


10681
CGCCGGCCAA AGAATTTTGT TATGGCCCGG ACTACTAACG AATCAACCTA TTTGAAAGAT


 


10741
AAAACCGGTG AGCGGCGTTT CTTACCTAAC ATGGCTGATA AGTCCCAAGC AATGGCTAAT


 


10801
CCGGTAACTG ATCTTGATGA TACGATGGTC AATCACATTT GGGGTGAAGC TGTTGGCCTC


 


10861
TACAAAGAAG GCTTTTCTTT CATATTGACG AAGGAGCAGC AGAAGCTCAT TGAGGATAAT


 


10921
CGAAAGATAT TTATGTACAT TGATGAAACT GAAAATCAAA TTGAACGGGT TCTCAGTACT


 


10981
TGGGACGATG ACTGGATTGA AAGCTCAGAA ATTGCTCATC AATTAGGTGA AGATAATCTG


 


11041
GTTAAGAATC GTTCATTAGC CAAAAAGATT AAGTATGTAA TGGATAACCG GCATGATTGG


 


11101
AAAGCAGGAC AACGGCGAAT TAAAGGAATT AGTCATCGTG GTTATAGAAA AGTGCATACA


 


11161
GATAATACAC TATGAATATA GTTAAGTGTA TGCAGAAAAA ACCTATTACA TCAACGTTTA


 


11221
TTAAGGTTTG CATACACTAC TACACTATTT TAATAATAAA AAATAAATAT ATATAAATAC


 


11281
TATATATGCG TTATAAAAAG TTGAAAGTTA GTGTATGCGT GTATGCAGTA ACTAAATCCG


 


11341
TTGAGAGAGT AAGAATTAAG CTGTATACAC TACTGTATGC ATAGTGTATG CAAGGAGGAA


 


11401
AATATGAATG AATTAAAAAA CATCAGCACA TCAGAATTAC TTGATGAACT GATTGAACGT


 


11461
AATGCATTAT TTCGTGTAGA TTGTGGTCTT TATCGGAATT GGGAGTTGAA GGGAAAATAT


 


11521
CAATTTAGCG ATATTAAATT GCCTAGCGCA TATCCTATTT ATGTAGGAAA TTCAATTATT


 


11581
GACCGCATGA TTAAGTGGGA GTGTGAGCAT TGACGAGTGA ACATAAAATT CAAAACGATA


 


11641
TCCGGGTTGC ACTATCAAAA CATCAGTGTA CAGTGTTCCG GGTAAATGTC GGTTCGGTAA


 


11701
AAACACCAGA TGGAAGATTT TTTTCAGCTG GTGTACCTAG TGGTCACCCG GACTTATATG


 


11761
GATTTCGTTG GTCGGATCAT CAAGTATTTT ATATTGAAGT GAAAAACGAA AAAGGTAAGC


 


11821
CTAGAGCGGA TCAAATTAGA TTTCATGAAA TGCTAACTAA ACGAGAAATT ATTCATGGAA


 


11881
TCGCTAGGTC TGCTGGAGAT GCAGTAAAAA TTGTTGAGGA AGGATTGATT GGTTATGGAT


 


11941
ATGAAAATAA AAAAGAACCA CCACAACAAG TAAAATTAGT TGAAGTAATT CAGGTTATAA


 


12001
CCTCTCGTGG AGCTGGAACA AAGGAGGATC CGATAAGAAA GATTATTCAG TATTGGAGCA


 


12061
AAGAAGGCAC ATTATTAGCA GAAAGTTTTG GAAACTAATT TTTCTTTGAG TCACGTTTCT


 


12121
CTATGTGCTT TAGTTGTTTT TCAGGAAGAC TATTATAAAA CTTATCAATT TGATTTGGCT


 


12181
GTGTTATTAA ATTGTCTGTA ATTATATTTA ACAGATTTAA AAGACTTTTC GCCATTTCTT


 


12241
GGTTATCAGT TAAGTCTATC TCACCAGGAT GAACAGCATT ATTTCCAACA ATTCTTACAA


 


12301
TATCAAACGC TTGTTGTAAT TTAGCCGGAA GTCCCTTAGA TACTAGATCA CCAATACGAT


 


12361
TATCTAATGA ATGTTTATTG GAAAATTCTT TTGTTAGTTT ATCTATTGCC AGACGAGACA


 


12421
AAGCAGCAGA TGCTCTTGGT GAATCTCTAA GTACTTTTGC CGCTTCAATA TAAATTTGTT


 


12481
TAATATCATC GGGCATATCT GGATTAGGAG CTGTTAATTT TAAAGGAAAA CTAGGAAAGA


 


12541
TAAGTATATC TTCATTGGAT TGCCGATTAT GAAACCAAAT AGAATCTAAA TAACATGAAG


 


12601
TGCATTTTGC AACTATTAAA TTATATTCGT ATTGTTTTGA TTTTATTTTT TCTGCATATT


 


12661
TCCAATCAAA TTGAGCATAT ACATGACAAT AAGGACAAAT AATAGGAGCT TTCCCTGTAA


 


12721
AATCATTACT AAAATATTTA GTCATTAAAT TCACCTCAAA TAAGATTATA GCGTAAAGGA


 


12781
GTAGATGTAA TGCTACACAA ATATAGAAAG TTACCAATTG TGGAAGCTGA ACAGTTTGAT


 


12841
GGGTCAGATG AAATGATTGA AAGGTATTCA GTGCATGTAT TTAATCCTAA TTTAGCTAAA


 


12901
AACATCTTCT TTATAGGTAT GAACGTTCTA GCTATTGGTG ATTGGATTGT TAAGGATGAA


 


12961
TATGGGAATT ATCAAGTGGT AGCTGATGAT ATATTCCGTA AAAGTTATGA GAGGTGCGAC


 


13021
TAATGCACAT TTATGAAGTA ATCGTTGTAG CTGTATTTGG CACAGATATT AGCCACTTTG


 


13081
TTGTTGCTAA GAATGCCGAT AATGCTAAGA AAATTATTCT TGATTATTAC AGCACTCGTG


 


13141
ATGATGGTAT CAGGCCAACT GTGACAATGT ATGACCTAAC AACAAAATTA ATCAATCTTA


 


13201
ATAACTACAT TGATGAGGTG ATGCTTGGAT GAGATTAAGT GACAAAATTA TTATGACTTC


 


13261
CTTTTTACTG TTATTAATTG TTTCAGTCGT CTTATCAATA GTTACGGGAA GTAAAGTCTG


 


13321
GATATGGATT TTTCTTTTTC TGATGACACT TGAGATGCTT TACAAAATTT GGCGTTAGGA


 


13381
GCGTGACTAA TGAAAGCAAG ATGTGGCGAT TGGAGTACTG ATGTTTATCC AGTTGAACCG


 


13441
TTTGTTTATG AAGTAGCAGC TGATGGTAAG AAAGAATTTT ATCAATCAGC ATGGGATGCT


 


13501
TTTAGTTCTG AATTGCATAT GAAAAATGTG ACGGCGAAGA TTATCAAAAT ACCAGTTGTT


 


13561
CCAATGAGTA ATGATGAAAT TAAAGCAGAG AGTGTTGCAT TTGAATTAGG CGAAAAAAGG


 


13621
CATTATCCAC ATGTGGAAGA ATTCACTAAT AGTAAACCGA GGAGGAAAGT TACATTTCTT


 


13681
CAGGTTGCGG AAGCTTTTGC AATTATAACG TGGATTATTT TAATAATTCT TATAATAAGT


 


13741
ACGGCGTTTA TGTAGGAGGT AAGTTACTAA TGACATTTGA AGAGGCGTTA AAACATGAAG


 


13801
AAAATAATGT GCCAGTATTC TATAGCGATC GAAAATATTA TGTAATCGGG CACAATGAGT


 


13861
TAACCGAACA ATTTACAATT CGTGAGCTAA GTGGTAATCC GTTATTTACG GTGCCGGTTG


 


13921
ATGTACAAGC GGAGGAATTG TCATGAGTAT TAAAATTAAT GCGCAAACAG TTGTATTTAA


 


13981
GGGACAAAGT TTTATTCCTA GCAAAAGTAA CTGTGAGTAT TGTCAATTTC CATTTAAGAA


 


14041
GTTAATGGTT ACTAAAGCAT CTCCAGATCC ATCAGTTAAG ACAGAAGTAC CAATCAAAGT


 


14101
TGACGGTGAG GTATTTAATT ATTGTCCTCA TTGTGGACGG AATTTACAAG GGTGAGGCAA


 


14161
TGTTACTTTT AATTGTATTA ATGATGATTG TCTTTATTGC TGGGTTCCTA TTGGGTAAGA


 


14221
AAAATCCATG ACGAAAAAAG GACCCACCGT AAAAGATGTG TCCTCACTAA AAATATTTAA


 


14281
CCATAATTAT TATATCAGAT AGCGAGGGTA CGTCATGCAA ACAAGTTTGA ATTTAGATAT


 


14341
TGATTGTCTG AAAACTGCAA GAAAGGTTAC TGACTTTCTT GATAAGAAGC TGGATCGCTA


 


14401
TCTGGCTTTA TCGGGGAAGC AACGTTTTGA TTTGAAGTCA CCAGGGATGG ACGGAATGCC


 


14461
TAAAGCTCCC AGCCATGGTA ACGGTAGTGA AAATCGAATG CTGAATATCT GGTTGGCAGA


 


14521
AGAGGTTGTT GATTGTGTGG GTTGCGCTAT GCGGAATATG ACAAAGGAAT CACAACGGAT


 


14581
TTTACTAAAT CGGTATTCGG ATCAGATGTT AACGTATAAC ATTGCTAGGG AGCTAAGTAT


 


14641
TAGTTCAGCA ACGTATAGTC GAAAGCAGGA AAAAGCGCTG TGTGAGTTTG CGGACCGGTT


 


14701
TGAATTTCAG TTAGTTAAGC ACGGTATTCA TACTGAAATT GATGATCTAC ACGTTTATCT


 


14761
TGACGAGGAA GATTGATAAA TTGATGATAG ATTATTGAGC GAGCAATTCA TGATAGAAAT


 


14821
GTGATAATAG TATTGTCGAA TGATTCGATA TTCATATAAT AATCTTCCCA AATGAAGTCT


 


14881
AGCTATTATG GCTAGGCTTT TGTATTATGT TTAATTGGGT GATTTTTATA TGATTGAATA


 


14941
CTTAAAAACA TTTTTGGAGG CTTTGAGTAT GAAGCCAAAA ACTAAATTTG TTGGAGTTAT


 


15001
TTTTGGAATA GTACTTTTAT GCTTAAAACC ATTTTTAATT CAATATAATA TGAAATGGTT


 


15061
TTACAATAAG TTTTCTTGGA TTATTATTTT AATTACTTTA TTTTTTGTAG CCTCATTAAT


 


15121
AATTGAAGTA ATAGTTGAAG TATACGAATG GGGTAGAAAC AAGTATAACA AACATAAAGT


 


15181
TGAAAGAGAT TACGAAAAAT ATATCTTAGG CTTGTCTGAT AAAAAGTTGG CAATTGTAAA


 


15241
GAAACTTTAC GCTAATGAAC ATCACCAAGG ATATTTAAGA CAAAACGATA CTAATGTTAT


 


15301
TGAATTGGTT AATATGTACG TAATTATGCA ACTTAATAAT GAAATTATAG TAAGAGAAAG


 


15361
CCAAGTTGAA GATATAAACG ATCCTGAATT TCTTTTTGTA TTACAGCCAC CGGCTTTACA


 


15421
CATCATAGAA AAGAATTCAG AAAAATTTAA ATAAATTTAA TTAGTTTAGC TTAACGGCTG


 


15481
ACTTTTTACT TTTTAGGAGG TGAGTAGCAT TACTCAAAAA TTAACACAGA AGCAACAACG


 


15541
ATTTGTCGAT GAGTACATTA TTTCGGGTAA TGCTACTCAG GCGGCAATTA AAGCTGGATA


 


15601
TTCTAAGAAG ACAGCTAAGC AGTCTGGTGC TGAAAACCTA GCAAAACCTT ACTTAAAAGC


 


15661
TGCAATCGAA AAACGCAATG CTGAAATTCA ATCCGAGAAA ACAGCTGATA TGACAGAGGT


 


15721
GATGGAATAT CTTACTTCAG TTATGCGTGG TGAGCAAACA GAATCGGTTG CTACTGCTAA


 


15781
GGGTATTTAT GAAGACGTTG AAGTGTCGGC AAAAGATCGT ATTAAAGCTG CTGAATTAAT


 


15841
TGGAAAGCGT CACGGCGCCT GGACTGATAA AAAGGTTATT TCTGGTGATG TTCAGATTGA


 


15901
TGTGGGAATG GGGGATTATG ATGATGAAGA GTGAAGAACA ATGGAGAAAA ATCAAAGATC


 


15961
ATCCTCATTA CTTGGTTAGC AATAAAGGTA ATGTTTACAG TGAGTATAAA GGCGGCTTGC


 


16021
TTAAACAGAT GAAAGATGCT TATGGATATT CTCAGGTTAA TTTAAACCGC CGCTCCAAAA


 


16081
AGGTGCATCG TTTAGTAGCG GAAGCTTTTA TCCCAAACCC AGACAAATTG CCTGAAGTTA


 


16141
ATCATAAAGA CGAAGATAAA AATAATAACC AGGTGGATAA CTTGGAATGG TGTACTAGCA


 


16201
AGTACAACAT GAATTATGGT GACGTGGAGA AAAGGTCAAT TCTTTCACAA CAAAGCCATA


 


16261
GTACTTGGAA AATTTATCAA TATGATTTAA ACGGTAATTT GGTAAAAGTA TGGAATTCAG


 


16321
CGAGAGAAGC CGACAGACAT GGATTCAACC GTAGAAGTGT GTATCGCTGT TGTGATGGGG


 


16381
AAATAAAATC TTTCAAAGGA TACATATGGT CAAGACAAAA GAAGGTGATA CCATGCCAAA


 


16441
CATCAAACTA AATTTTCCTA AACCATACAA CGTTTTCAAT AAACAAATTT TTGATAACTT


 


16501
GTTTGATTAC AGTCATTTCG TTGAGGTTTG GTACTGACTT ATGGCGGTGC ATCTTCTGGT


 


16561
AAATCGCATG GTGTGGTACA GAAAGTTGTA CTTAAATCAC TCCAACACTG GAAACATCCC


 


16621
CGCAAAGTGC TATGGCTGCG GAAAGTTGAT CGAACAATTC AAGAATCTAT CTTCGCTGAC


 


16681
GTAATTGACT GTTTATCAAA TTGGCAATTG TTATCGTTAT GTAGAGTAAA TAAATCAAAC


 


16741
CGTACTGTTC ATTTACCGAA CGGTGCGGTT TTCCTGTTTA AGGGTATGGA TGACCCAGAA


 


16801
AAGATTAAGT CAATCAAAGG GTTATCTGAT GTGGTAATGG AAGAAGCTTC CGAATTTACA


 


16861
CAGGACGACT TCACGCAGCT TACCCTACGT CTCCGTGAAC CTAAGCATAA GAAACGACAA


 


16921
TTGTTTTGTA TGTTTAATCC AGTTAGCAAA TTGAACTGGA CTTATAAGCA ATGGTTTGAT


 


16981
CCGAAAGTGA AAGTTAATCC GGAACGAGTA TCAATTCACC AATCAACTTA CAAGGATAAT


 


17041
CACTTTTTGG ACGCTGATAA CATTGCAACG ATTGAGAACT TAAAACAAAC CAACCCGGCC


 


17101
TACTATAAAA TCTATACGCT GGGCGAGTTT GCTACATTGG ATAAGCTGGT CTTTCCAGAG


 


17161
TTTGAAAAAC GTCGGTTAAG TATTCGAACC TTATCACAGC TTCCCTCGTA CTTCGGCTTG


 


17221
GACTTTGGGT ACACTAACGA TGAAACAGCC TTTATGCACG TTAAAGTGGA TGAGAGTACC


 


17281
CGTAAAATTT ACGTGATGGA AGAGTACGCT AAGCACGGTA TGTTGAACGA TGATATTGCC


 


17341
CGAATAATTA AACAAATGGG TTATAGCAAG GAAGTTATTA CTGCTGATGC TGCTGAGCCT


 


17401
AAATCGATTG CTGAAATTAA ACGCGATGGT ATCTCGCGGA TTCGTCCAGC TAAAAAGGGG


 


17461
AAGGACAGCA TTATACAGGG ACTTTCATTT ATGCAGCAAT ATCACTTAGT CGTTGATGAC


 


17521
CGGTGTGTGA AAACGATTGA GGAATTGGAA AATTATACAT ACAAGAAGGA CAAACAAACT


 


17581
GGTGAATACA CCAATGAGCC TGTCGATAGT TACAACCACG AAATCGATGC TATCAGGTAT


 


17641
GCTTTATCTG AAATCAACGG AATGGCTAGT CCAAAGGCAA CTGTAATGAA AAATATTTAT


 


17701
ATTTAGGTGG TGATTGAATG GAAACAGTAA ACGGTAAAGG ACAAATTTTA GATGGCCATA


 


17761
TTTTTATCTA TCCAGCTGAT GAAGAAGAAC TTGATCCGCA TGATTTACTG TCGTTCATGA


 


17821
GAAGAAATAT TCAGTATGCT AAGGATTACA AGCATAATAT GCAAATGTAT CTAGGTAATC


 


17881
ACGATATCTT AGATCAACAG CGGCGGATGT ATGGGCCAGA TAATCGGCTA GTAGCAAATT


 


17941
TACCGCATTA TATTGTTGAT ACTTATAATG GATTCTTTAC TGGAATCCCA CCTAAGATTA


 


18001
CTTTAGATGA TAAGAATGAG AACGAAGCAT TACAGCAATG GAATGACACT AATTCGTTCC


 


18061
AGGACAAATT GAGTGAAATT AGTAAGCAAA CGGATATCTA CGGACGTTCG TTTGCTTTTA


 


18121
TTTATCAAGA TGAGAACGCA GACACTTGTA TTGCTTATGC TTCTCCTACA GATGCCTTCA


 


18181
TGGTTTACGA TGATACGGTT GCTAGAAAAC CTTTTGCTTT TGTTCGTTAC TGGAAAGATA


 


18241
CTGAAAGCGG ATTATGGACC GGAATGGTTT ATTACGCTAA TAAAATTAAA ACCTTTAAAG


 


18301
GTAGTGTTGT TGAAGATTCA GATCAAAATA ATATGTATAA TTTAGTGCCA GCAGTTGAAT


 


18361
TTTATGGAAA TGAAGAGCGG CAAGGTGTTT TTGATAATGT GAAAACCTTA ATCGACGAAT


 


18421
TAGACAGAGT GTTATCACAG AAAGCTAACC AAGTGGAATA TTTTGATAAT GCTTACCTTA


 


18481
AAATTCTTGG TCTTGATTTA GATGAGGATG GTGATGGTAG ACCGGATGCT AATTTAATTG


 


18541
GTAATCAAAT GATTTATTCG CCTAATGCTG ATGCTGCTAA TGCCGATGTC GAATTCATTT


 


18601
CAAAACCAGA TGGTGATAAT ATGCAAGAAC ATATTATTGA CCGGCTTGTT TCAATGATTT


 


18661
ACCAGGTAAG TATGGTTGCT AACCTTAATG ATGAAGCGTT TGCTGGTAAT AGTTCTGGGG


 


18721
TGGCTTTGCA ATATAAGTTA CTTCCAATGC GAAATATGGC AGCTAATAAA GAGCGTAAAT


 


18781
TTACTCAGGC ACTCCGGAAG TTATATCGAA TAGTGTTTAG TGCTGATCAA GTAGTCAAAG


 


18841
ATAAGGAAGC CTGGCAAGAC TTGCTCTTTG ATTTCAAACA AAACTTACCG ATTGATGTTT


 


18901
CTGAAGAAGC TGATACTTTA CAAAAACTAT CAGGGGTTGT GTCAAAAGAA ACTGCATTCC


 


18961
GAAATAGTCG TTTAATTGAT GATCCTAAAA AAGAAGTTGA GCGTATGCAA AAAGAGAAGC


 


19021
AGGAAGAAAT AAACCAAGCG CTTCAACATT CTGCTTCTGC TACAGATCAA ATGCTAATGG


 


19081
ATGATCAAAA AGAAAATGAT AAAGAGATAG TTGGTTTCCG GAAGAACGGT GAATCCGATG


 


19141
ACGAAGAAGA ATAATTATTG GGCTGATCGT ATTGCTCGGG AACGTAAATG GCAAGAAGAG


 


19201
CAATTAAGTA AAGATGCTCA ATTTAATCAG CGCCTTCAAC AGTATTATGA TCAAGCAATT


 


19261
GTCCAGATTA ATAAAGACAT TGAAGATCAG ATAAATTCTT TAGCTGTCCG GAATAAAGTT


 


19321
TCTTATGCTG AAGCTCAAAA AGAAGTGTCC ACTACCGATA TTGCTGATTA TGAAACAGAA


 


19381
GCTAAGAAGG TAGTTCAGGA AGCTAATCGT TTAAGAGCAC AAGGGAAGCA TGTTACTTAC


 


19441
AATGATTTCT CTGATGAAGT TAATGAACGA TTGAGGAATT ATAATACGGC GATGCGATAT


 


19501
AACCGATTGA ATTTATTGAA ATCTAAAATT GGTTTATCGA TGGTTGAAGC CGGAATGAAT


 


19561
ATTGATGCTG ATATGCAAGC TAAAATTGGT AAAGATTATA CTGATGAGCT AAAACGTCAG


 


19621
TCTGGTATTC TAGATCATTC TACCGAAAAT AGTTCGTTTT GGACTTCTAA AGATGTTGCG


 


19681
AAACAAGTAA TGAAGCAAAT TAATGGAGCA ACTTTTAGTC AACGAATTTG GGCTAATCAA


 


19741
GATACTTTGA AAGCTCAACT TGATACGGTT ATCACCAACG GAATTTTAAC TGGTAAGAAT


 


19801
CCGCGAGTTG TAGCAAGACA ATTAAGAGAT AAAGTAAAAG TCACTGTTAA AAATCACAGT


 


19861
TATGTTACTG AACGTATTGC CAGGACAGAA TCAGCACGGG TTCAGTATTC TGCTCAGATT


 


19921
GAATTAATCA AAAAGAATGG TTATCAATTT GTCCACTGGA TTGCGGAGCC AAGAGCCTGT


 


19981
GATGAGTGTC GAAAGATTGC GACGCAAGAT AATGGCTTTG GTGATGGTAT TTATCGAATT


 


20041
AATAAAGTTC CTAAAATACC AGACGATACT CATCCTAATT GTCGCTGTTC AATTAGTGAG


 


20101
ACATGGGTCG ATGGTCAACG CAATATAGCA TTATCTGATG ATGAACAGGC GGCATTGAAT


 


20161
AATTATATTA GTTCAGATTC ATATAAGATT AATGATGATT TGAGGCGTAA TAAGATTTCT


 


20221
AAAAATAAGA AACAATTTAT TGAAAACTTA GATGCCGCAT TGGCTAAAAT GCCAATTTAT


 


20281
CATAGTAGCA AGCCACTCCA GCGTGATTAT TTCTTTGATA AACAAGAAGC ATTGGATGAT


 


20341
TTTATTAGTA ATTTTGAAAT TGGTGGAGTC TTCACTGATT CATCATACAT TTCAACTTCT


 


20401
AAAATTTATT ATGGACAGGG CAAAGAGACA ATTCATGTTA TTATTAAATC AAGTAAGACA


 


20461
GGAAGAGATA TCTCTGAGTT TAATTTTAAT GAGCAGGAAG TATTATTCCC CAGAAATAGT


 


20521
AAGTTTAGGA TTGATGATGC ATACGTTGAT GATAACGGGA AGATGACAAT GGTTTGGAGT


 


20581
GAATTAGATG AGTAACAAGC CTTTTACTGA TAAACGTTGG CGAGATAATT CTTTGGAAGG


 


20641
CGTTAAATTT GATAATTCAA AAGTAACGTC AGAACAAAAG AAGAAGACAG AAGAATTCCA


 


20701
TGAGTTATTC AAAAAGACGT TTGCTAAACA ATTAAAAGAA AAACACTCAC ATAAAAAGTA


 


20761
GGTGATCCAA TGGGAAATAA TGATTTCTTT ACGGTAACGT ACAAAATACT AAGTTATCTT


 


20821
AAGTATTGTT ATGAAAATGG AATTAATCCT GATCCTAATA TTCTTAATGC TGATACATTT


 


20881
AATATTAGTA AAGTTCAATT TGGAAGAACT CTACAAATGT TAAGCGAGCA TGGTTATATT


 


20941
TCAGGAGTGA GATTTACACA AGCCAAAATC GAAGGTACTG TTGTTGGTGG ACTCCACAAT


 


21001
ACGTCAATAA CGGTTGAAGG TCTGCAATAT TTAGCTGAAA ACTCAATGAT GAAAAAAGCA


 


21061
TATCGAATTT TTAAAGAAGT CAGAGATTGG CTTCCGGGTT TCTAAGCATT CACAAATTAG


 


21121
TGAGTGCTTT TTGTTTTGGA CTTTTTACTT GTTGCAGTCG TTAAAGAACA ACCCGGATAT


 


21181
TACAGTCCAC CGGACTATAA ACGAGGTGTA TTTATGTTTG AAAAATTACC AATGCGTTTA


 


21241
CAATTCTTTG CTGAAGATCC AACGCCAGAT CCAGATAATG ATGGTGCACC TGAAGGAACT


 


21301
GATGATGGAG ATAACGGTAA AAGTGAAAAG ACATTTACTC AAGCAGAATT AAACGATATT


 


21361
GTCAAAGCCC GAGTCAATCG AGCCTTGAAG AATAAGCAAG AGGAAATTGA CCAGGCTAAG


 


21421
AGTGAAGCTA CTAAACTTGC CAAGATGAAT AAGGATCAAA AGCAAGAATA TAAGCTTCAA


 


21481
CAAACTGAAA AACGTGCCCA AGATGCTGAA GCAGAATTGG CCCGTTATAA AATGCGTGAT


 


21541
ACAGCGAAGC AACAATTAAT TGATGGCGGT TATGACAATC CAACTGATGA AGATATCGAT


 


21601
TTAATTGTTA CTGATAAAGC AGAAACAACT AAAGAACGTG GTGAAGCATT TCTTAAAGCT


 


21661
TATAACCGAA TTAAAGAAAA TGTTCGTCAA GAACTATTAA AGGGAAAGTC ACCACGAATT


 


21721
AATGGTGCTC CTGCTACTGC AATGACTAAA GAACAAATTG CAAAGATCAA GGATCCCGTC


 


21781
AAACGGGTCC AAGCCATTCG GGATAACTTA TCCCAATATG AAAAATAAAA GGAGGAATAT


 


21841
AAAATGGCTG AAACTAATTT AACGACAAGT ACAGACCTAG TTGCACAATC TATCGACTTT


 


21901
GTAGAACAAT TCTCTGGAGG AATCCAAACT TTATTGAATG CATTGGGAGT TATTCGTATG


 


21961
CAGCCAATGA CTACTGGTTC ACAGATTAAG ATTTACAAGT CAGAAGTAAC TAAGGTCGAT


 


22021
GGTAATGTTG CTGAAGGGGA AGTTATTCCG TTAAGTAAGG TTACTCGCAA GCTAGCTAAT


 


22081
ACTTTGACAT TAGGATTTAA AAAGTATCGT AAGGTAACTA CCATTGAAGC TATTCAGTCA


 


22141
GCTGGTGGTG CTACACCTGC TATCGTGGAT ACTGATAATA AGCTACTTCG AGAAATTCAA


 


22201
AAGGATGTTA AGAAGGACTT ATTTAATTAC ATTACCAAGT CTGATGCAAA CAAGACTACC


 


22261
GCCTCTGGTG ACGATTTTCA AAAGGCAATG GCGGCAGCTT TAGGACAACT TTCAGTTAAG


 


22321
TGGGAAGACT ACGACACACA AACTGTCGCC TTTGCTAATC CGCTTGATCT ATATGCATGG


 


22381
TTAGGTAACC AAACTCTTAC TGTTCAATCT GCCTTTGGTT TGCAATACAT TCAGAATTTC


 


22441
CTTGGCTTTG ACACTATTAT TCTAAGTGCT GAAGTACCAC AGGGAACGAT TGCTACAACA


 


22501
GTTGCAGATA ATATCAATTA CTTCTACGCT CCAATTTCAT CCGTTGGTCA GTTATTTAAC


 


22561
ATGACTTCTG ATGAAACTGG TTTAATTGGT GTAACTCATG ATGCAGTTAA TAACAATTTG


 


22621
TCATACGAAA CTGTTGTAAC AATGGCTAAT GTATTGACTA CAGAACGTTT GGACGGCATT


 


22681
GTATTGTCAA CAATCAGTGG TGCTAAGTCT GCTGGTAAGT AGGTGATTGA ATGGACCAGA


 


22741
ATACGGTTTT GCAAAACTTA AAAGTAATGC TTGAAATTAA AAATGATGAC CGTGATGCTT


 


22801
TGCTGAAACT AATCATTGAT AATACAGACC AAGCATTGCG ATTTAAGCTA GAACTAACTG


 


22861
AAGACGAAAA TTTACCTGGA GAACTAGGTT ATATTGAATT AGAAGTTTCA GTTCGACGAT


 


22921
TCAATCGACT GCAAAACGAA GGGATGAGTC AATATAGTCA AGAAGGGGAA AGTATTACTT


 


22981
TTAATTCTTC AGATTTTGAT GATTTCCTTG ATGATATTGA TTTGTGGAAA CGACGACACC


 


23041
AGAAAGATGT TAAATCTTTA GGTGCCGTTT CTTTTATTAA TCCTTATGCG GGGATGAGTA


 


23101
AAAATGCGAA AAACACAGAT AATTAAGTTT TATTTTCAAG ATGAAAATGG CTATAATCCA


 


23161
TATGCAGAAG AAGACACGAT TACTAGTCCT AAGCTAGTAG CACAACGGTA TGCCAATGTT


 


23221
ACTGATGTAG GGACAAATCG CCTAGTTGAA TTATTTAGCA GGCTAGATCA GAACGCCAAG


 


23281
GTAATTCGAT TGGAGTCTCC AGTAAATGAC TCTTGGTCAT ATCTGACTAT TGATGATTGT


 


23341
CCTATCAAGT ATCGTCTTGA AACCTGTCGG AAACCATTAA AAGGCACAAC GCTGATTGTA


 


23401
GGTGAAGCCA GTGGCTAATT CTTTTAAGGT TGATGTTAAA GGTACTAAAG AACTAGCGAA


 


23461
TTTCTTAAAG AAGAATAAAG ATTTGACTCC AGTTAAACGG ATAGTCGCAA AACATGGAGC


 


23521
AGGTCTCAAA AAGCAAACGC AGCAAAATAT GAACAATTTG TATAAGGGCC ACTACGAATG


 


23581
GAAGAAGGGG GCTGGGCTAA CAATGGTTAG CCCTACCGGG AATACTAGAC GGTCCGTAAC


 


23641
AAATACAATT TCTAATAATG GTTTAACAGC AACGGTTGCT CCACAAACTG AATATTTCCC


 


23701
ATATCTTGAA TATGGAACTC GCTTTATGGC AGCACGACCG ACATTACATC CAGCGTTTGC


 


23761
AATTGAGTCT ATGAAATTTG CTAATGATTT GAATAAGTTA TTTAAGTAGG TGAGAAAATG


 


23821
TCACCAAGTA TTGAAATTTA TGATGCAGTT TTTGCTCAGG TCCAAAAGCA TTATCAGACC


 


23881
TACGATCACC CACCGCAGTT AAATGAACCA GTTACCTATC CATTCGTGGT TGTTGATGAT


 


23941
AGCCAGTCGA TTTTGACGAA CTATAAAACA GCCACAGGAA TGCGGGTAAC TTTAATAGTC


 


24001
CATGTGTGGG GGAAGTCTAA CCAACGTAAG ACTGTTACTA AGATGGTTGA TGAAATTAGT


 


24061
CGTCTGGGGA TGCAAGCAGT TCGGACGAAA CATTATGCTT GGCAAGGACG ACCTAATGAG


 


24121
CAAGAACAAC AATTATTAAC TGATACGAGT GTTCCGAATA CTGTGTTAAA GCACGGTTAT


 


24181
TTAACACTCG TTTTTGATTT GAAATAAAGG AGGATAAATA TGGCAACATA TCCAGTATTG


 


24241
GAAGGGAAGA ATGCAGTTCT TTTTGAACGA CTATTAGAAA ATGCAAGGAA AGAGCCGGCA


 


24301
CAATTGATCC CGTATCAAAC ATCACTAAGT TATGATCCTA AACGGGATAC TGATTCAACA


 


24361
ACTACGAAGA TGGGGAATGT TCCTACTGCT TCTAATATTG AAACAGATTT AGAAGTAGAG


 


24421
TTCCTAAATG CAATTTCCAA AGCTGCAGAT GATGTTTATG ATTCTTTGTA CTTTAATAAG


 


24481
AAGATTGAAG TATGGAAGGT TCATATTGAT CGAGTCCGGT CAGATGGCAA AGTTTATGCC


 


24541
GAATATATGC AAGGAATTGT GTCAGAAGAC TCTAATGATA ACGACCCAGA CGATCATTCA


 


24601
ACTCGGGATG TGACCTTTAC GATTGATGGC GTGGCCAAAC GTGGGTGGGT CACTCTACCG


 


24661
CCAGAAATCA AGGAAGAAAT TGACTATGTA TTCCGTGGCT TGGCACAGCT TAAAGGCGAT


 


24721
GACGACAACG GTGAAGGTGA AGCTTTTGCT GATGGCGATC GTGGTGCTGG TGCAAATGAA


 


24781
GCAGTAACAA CTGAATAGGA GGAAGATTAT GAAGTTAAAA ATTAATGGTC AAGACCAATC


 


24841
ATTTGTATTC GGAGTTAAGT TTTTGCGAAA ACTTGATGCT TATCGGGGCG CTGAACAAGA


 


24901
AATCCAAGGA GTTAAGGTTA AGCTAGGAAT GGGGCTAGCC ATGATGCTTC CCCAATTAAT


 


24961
GACTAAGGAT GCGGCCGCTT TGGCAGACGT GTTGTACTGT GCGGCTAAGT CTAGCATTAA


 


25021
GTTAGATACA ATTGATGATT ATATTGATAA TTGCAAGGAC TTGGATTCAT TATTTAATCG


 


25081
GGTAATGAAT GAAATTAAGG CAAGTAATGC CGCTAAGCCG ATTGCAAAAA ATCTAAAAGC


 


25141
CTAGATGGTC CTGAGCTTAG TTCAGAACAA AGCTATCACG AAATTCTTTT GAATTCGTTG


 


25201
GCTTATCTAG GCTTTCATAA TATTTCAGAA ATTGAAGAAA TGGGATTGGC TGAATATCAG


 


25261
CTCCGGATGG AAGCCTATAA CCTCCAACGG GTTAGCCAGG AACGAGACTT AGCATTGCAA


 


25321
GCTTTCCTTA ATCAGTCGGT ACAAGCGACG AAAGGGAGCG AAAAGCACCC AATTCCGAAG


 


25381
TATAAGAAGT TTAGCCAATT TTTTGATTAT GATAAATTTG TTGATGATGT TCGTGGGCAC


 


25441
TATGAGCCTG ACTATCAGCC AACAAGCAAG GCCAGCCTTG AAAAGAAACG AAATGATCTA


 


25501
ATCGTCAAGC GGTGGCGTGA ATTCCGGAAG ATGAAACAAA AACAGAGAGG AGGTAATGGC


 


25561
TAGTGTCACA ATCAATGAGA GTTGAAGCGG TATTATCAGC ATACGATGAG AGTTTTAGCG


 


25621
CAACCTTAGA TAAGGCGCTT AAATCGATTA ATAATTTAGG CCGTGAAACC CAGTCAACCT


 


25681
CTCAAACTGT TAGTGCAGGT GGTTCTAGTA TTTCCAGTAC CTTTAAATCG ATGGCTGGAG


 


25741
CAATGGGTGT AGTTGCGATT GCTGGTAAAG CATGGGACGT TGTTAAAGAT TCAATGAGTG


 


25801
GCGCCATTAA CCGGTTTGAT ACATTAAACA AGTATCCGGT AGTAATGAAG GCTTTGAATT


 


25861
ATTCAACTAA GGATGTTGCA AAGTCAACCG CTATCTTATC TAAGGGAATT GATGGATTAC


 


25921
CTACTTCTTT GCAAGACGTT ACAAGTGTTG CCCAACAATT AGCGCCATTA ACTGGTAGTG


 


25981
CAACTAAGGC TTCTAAGTCG GCGATTGCCT TGAATAATGC CTTCCTTGCC TCCGGTGCTA


 


26041
GTGTTGCCGA TACCTCTCGT GGACTTCAAC AATACACACA AATGCTTTCA ACTGGTAAAG


 


26101
TCGATTTAAT GTCTTATCGA ACATTGATGG AAACCATGCC AATTGCATTA CGTAAAGTCG


 


26161
CCAATTCATT TGGTTTTACT GGTAAGTCTG CTGAACAAGA CCTTTATAAA GCTTTGCAGT


 


26221
CAGGACAAAT TACGGTAGAT CAGTTGAATG ATCGTTTTAT CAAGCTGAAT GGTGGAGTTA


 


26281
ATGGTTTTGC TCAATTAGCA AAGAAAAATA GTGAAGGTAT CGGTACATCT TTTGCAAACT


 


26341
TAAAAAATGC CGTTGTCAAA AATCTGGCAA ATATGTTATC GGCAATTGAC AATGGTTTTA


 


26401
AGCAAGCGGG CTTTGGAAGT ATTGCACAAG TCCTAGACAA CATGAAGGGT AGTATTAATT


 


26461
CTGCTTTTCA AGTTATTGGA CCAGTTGTTA CTAATGCTAC TGTTGTAGTT CTTAATTTTG


 


26521
CAAAGGTTAT AGGCGGAGCG CTTAAATCTG CTTTCAGTAA TGATATTTTT AAAACAGCAG


 


26581
TTGTGGGAAT ATTAGGCTTT GTGGGTGCAG TTATGGCAGC CCATAAGGTT ATTTCAATAT


 


26641
TTACAACATT AAGATCTGCA ATAGTTGGTT TAAGTGTGAT TACAAAAGCT GGTAATTTGG


 


26701
CAATGGCGTT TAGTGAAGCA ATGTCAACAC TTGCTAAAAC TTCTAAGATT GCTGGTGGAG


 


26761
CGATGAAAGC ATTCAGTGCG GTGGCCTCAT TAGGTCCCTG GGGAATTATT GCTGTTGCAA


 


26821
TTGCAGCTGT GGTTGCAGCC TTAACTTATT TCTTTACCCA AACGAAAACC GGTAGGGCTT


 


26881
TATGGCAAAG TTTTACTACG TGGTTATCTG GAGTATGGCA GAGTTTGGTT GGAGTGGCTA


 


26941
CTACTGTTTG GAATGCAATT GGTAATGCTA TTAATGCAGT AGTTAATTTT ATTAAACCTT


 


27001
ATTGGCAAGG ATTATTAACA TTCTTTACCG GAATCTGGAC ATCAATTGTG GCGGGTGTTG


 


27061
CTCCAATTTG GCAAGGGTTA GTTAATGTCT TTAATAGCAT TATCAGTGCA ATTTTAGCCG


 


27121
TTTGGCAGGC TTTAGCTCCA ATTATTGTTC CGATTGTAGC TGGTGTAGTT GCTATCATTG


 


27181
GGGCAACCCT AATTACGATT GTTACCGTCT TTCAAACTGT GTGTAATATG CTTGTACCCA


 


27241
TTGTTCAAGT TGTATGGCAA TTAATTTCAA CAGTTGTATC TACTGCTATT ACGATGCTAG


 


27301
GTACAATAAT CCAAACAGGC TTGGCAATTA TCGTTGCTAT TTGGAATGTG GTCTGGAATA


 


27361
CATTCAGTAT TGTTGTAAGT ACGGTATGGA ACGTTATTAC TACTATTATA TCTACCGTGC


 


27421
TGAACGTTAT TGCGGGAATA ATTCAAGCTA TCACTGCTGC AATACAAGGC GATTGGTCAG


 


27481
GAGCTTGGAA TGCAATTCAG AATGTTGTAT CAACTGTTTG GAATGCAATT GCCAGTATTA


 


27541
CTTCTAGTGT ATTGAATGGA ATAAAAGGAA TCTTTGATGG TGTAATGAAT GGTTTAAAGA


 


27601
GTATTACTTC TAGTAGTTGG AATGGTATTA AATCGCTATT CAGTGAAGGT GTTAATTTCA


 


27661
TTAAATCAGT TGTTCATATA GATTTAGGTG CTGCTGGTAG AGCTATCATG AATTCACTTT


 


27721
GGAATGGAAT GAAATCCATT TGGAATAGTA TTAAGAATTG GGTTAGCGGT ATTGCTGATT


 


27781
GGATTAAAGA ACATAAAGGA CCAATCAGTT ATGACCGTAA GTTACTTATT CCAGCTGGGC


 


27841
AAGCAATTAT GAATGGTCTT AATAACGGAT TGATTAATGG ATTCAGTGAA GTTCAATCAA


 


27901
ACGTTAGTGA TATGGCTAAT CAAATTCAGC AAGCTATTAC TAATCCAGGC TTTGATATTG


 


27961
GAGCAAGTAT TGGTAACTTG GGTTCAATTA ATTCAAATTA TACTGGTAGC CTGGCAATTC


 


28021
AAGATAGTCA GTTACAAATG CAGAATAATG CTTTGCTTCG TCAATTACTT AATAAAGACA


 


28081
CGACAATGGT TCTTGACGAT GGCACTCTTG TTGGCTATAC AGCGGATCAA TACGATTATC


 


28141
GCTTGGGTCA AAATACAGCA TTGAAGGATA GGTGGAGCCG ATGAAATTCT TAAATAATGA


 


28201
CTATTCTTTT CGTGGATTAG GACCCACAAA GGATGATCCA GAATACTTAG AAAATGCAGA


 


28261
ATATATCGAC TTTGCCGGTT TTAATTCTTC TGATTATGAT TGGTGGTTGA TTGATCGAAC


 


28321
AGCAACTACG CCAGAAGAAC AAGAAATTAC AGAAAGCGTC CCTTACATGC AAGGAGAATA


 


28381
TGATTTCTCA ATGTATGATC AGGAACGTTT TTTTAAGACC CGTGAGTTGA CTTATAAGTT


 


28441
TGTATATTTT GGTGAAGTTT ATCAGGATCG TAAAGCTTAC GAAGAGGAGC TTAAACGGCA


 


28501
ATTACTGCCA CATGGTTTCA CTAAACTAAT TGATTCTCAT GATCCTGTTT ACTACTGGTC


 


28561
AGCTAAGTGT ACTAGTGTTG AGGTTGAGGA TGACCAAGAA AAGGGAATGC TCACAGCAAC


 


28621
TATTACTTTC AAGGCTTATC CTTTTGCTTA TACTAATCAT AACGAGGGCA CCGATTATTG


 


28681
GGATGATGTC GCATTTGATC ATTGGATTTG GCAACCAGTT AAATTCAATG TTAACGGTGA


 


28741
TCAGGATGTT AATGTTAAGA ATATCGGCTC ACGACCAGTC GAATGCTCAT TTCAATTGAC


 


28801
AGGGTCCATA ACTTTGAAGA ACGATTCAAT TGGTGAAGTA GGTTTAACTC AAGACAATTT


 


28861
TAAAACAACC ACGATTGTAT TAGAGATGGG TGACAATAAA ATGCATCTAT CCGGAAACGG


 


28921
GACAATTGAA TTTCAATTTA AGCGTGAGGA GATGATTTAG TGTACCGAAT TATTGGTTAT


 


28981
AATGAACCAA CAGATAAAGC AGGATTTATT GTACTGGATC CCCGAGTTAA TCGTCATATT


 


29041
AGTTCGGGAA AACTCACGCT TAAAGAATCT AATATTGATG ATTTGACTAT TACGGTTAAT


 


29101
CAAGCAAGTC CATTATGGGA CAACGTAAGG CCTTATCATA CTCATGTTAA CGTTTATGAT


 


29161
GATAATGAAC TTATTTTTCG TGGACGAGCT ATCAAACCTA AAAAGTCGAT GGAAGAAAGC


 


29221
GGACAATTCA TTCGTGAATA TGTTTTTGAA GATATTGAAG CATATCTCAT GGATAGCACC


 


29281
CAAAGATTTT ATGAAGGTGT TGGTCAAACG CCCAAAGAAT TTTTACAAAC TTTAATCGAT


 


29341
GTTCATAATT CACAGGTTCC TGACTATAAA AAGTTTCAAG TCCGGAATGT AAATGTCACT


 


29401
AATAATAAGG ATGACCAATA TCGACAAATT GATTATCCCA AAACTAGCGA TGCTATTAAT


 


29461
GATAAATTAG TTAAATCTCT TGGTGGTTAT ATTGTGACTA CTTACAACGC TAACGGAATA


 


29521
AACTACATTG ACTACTTAAC GGATATTGGG GTTGATCATA AAGATGATAC TCCTATTCAG


 


29581
TTAGCTAAAA ATATGAAGTC TGCAAGTATG CAAATTGATC CTACTAAGGT GATTACAAGA


 


29641
CTGATTCCAC TGGGAAAGAC ACTAGAACCA TCAAAAGTTG ATGTAAGTGA TGATGATGGA


 


29701
GAGGGCGGTT CTGGATCATT AGATAGCCCT GAAGAATTTT GTAAATCAGA AATTAATGCT


 


29761
ACTTGGGGTA GTGATATTAA TAATATGAAA CAAGATTTTG CCGCTCGTTC TTCGAGAGTT


 


29821
CGGGCTTGGG GAGTGGACGT TAATCGTTTA TATGATGTGG TGAAAAATGC TGGAGTAAGT


 


29881
CCTGAATGGT TCTTTGCTTA TGAACTTCAA GAACAAGGAA CTTACTATGG ATGGCTTAAC


 


29941
CATACTTATC GACACGGTGA TGCGTATAGT GATGCGCAAT CTGTTTGTGA GTGGATTAAA


 


30001
AATTGTTCAA ATAGTAATTC CATTAATCCA GCATGGAGCG CACCGGAAGG ATCAATGGCG


 


30061
CCGAATCAAG CATTAGCGGA TAAATGGAAT CAAGAGTTTG GAAAAGGTAC TATTGGCCGC


 


30121
GTTTATTTAC AAGGGACTGC CGCTGCTGTT TGGGATTTAG CTGGTCAAAC GCCTAATCCA


 


30181
GCTATTGGAA AGCCAATTAG TGGATGCATT TCTTGTATTA AACGTTGGGG TGGTCATTCT


 


30241
AATGCAGCTG GTGGTACATG GGGATGGCCT TTTCCTGATG TTGGGGAAGG TCATTTTTCT


 


30301
CAAGTTCAGA GTTTCGGAAA TGATGGCGGA TATCGTCAAA ATAGTTATCA CGATGGTGTG


 


30361
GATTTTGGAT CAATAGATCA TCCTGGTAGA GAAGTGCATT GTATTCATGG TGGAACGGTA


 


30421
ACTATCAAAT CAGCTATGGG TGGCTTAGGT AATTTTGTGG TTATTCATAC GCCGGAAGGA


 


30481
TTCAATATCG TTTATCAAGA AGCTTTTAGT TCTCCCTCTA ATATTATTGT TAGTGTTGGG


 


30541
CAAAAAGTAA AAACTGGTGA TGTAATTGGA TATCGTGATA CAGACCATGT TCATATTGGC


 


30601
GTAACTAAGC AAGATTTTTA TCAAGCAGTT CGAAATTCTT TTTCTCCTGC AGGTGGTTGG


 


30661
CTAGATCCAG TAAAACTAAT TAAAGAAGGT GGCGATGGGT CTAAACCACA AGAAGGAAAG


 


30721
AAAGATCAAA CTGTTGATAA TAGTAATGCT GCACGTCCTA AATTAACCAT TACTACTGTC


 


30781
AATAACGGTA GAGACTATAT TGATATTCCT GATTTACAAA AAGAATTCGG TATTATTGAG


 


30841
GGAACTGTTG AATTTGATAA TGTAGATGAT CCGAATGTTT TAATGCAACA AGCTCAAACA


 


30901
TGGATAAAGG CTCAAAGAAT ACCTCAAAGT TGGGAAGTTA CAGCTTTAGA ATTACATATG


 


30961
ACAAACTTCA AATCTTTTAA GGTTGCTGAT AGGTACATGT TTATTAATCC AAATGTTGCA


 


31021
AAACCCCAAT TATTACGAAT TACTCAAAAA GAAATTGATT TACTAAAGCC CCATGCGTCT


 


31081
TCATTAACGA TTGGTGATAA GACGATGGGG CTTACTGATT ATCAGTTAGA AAATCAAGTC


 


31141
AATTTTCAAC AATTTAAGGA AATTCGAGTG ATGGTTAATC AGGTTGTCCA AACCCAAGAG


 


31201
CAATCTGCTA ATAACAATAA TAAGGTTATG CAAAATTTTG CTAGTAGTGC TGATCTTGCA


 


31261
CAAATGAGAC AGGATCTAAG AAATCTTCAA GATGATAACG ATCGTGCTCG CAAAGGAATG


 


31321
GTTTCCTTAG AAGAATTCAA TAAACTAAAG GAACAAGTAG AAAAACTAAC AACAGGAGGC


 


31381
GATGATAATG GCAAGTGAAA CCTATGATTA TGAGTCATTT GATAATACGG ATCATACTAT


 


31441
GAAACAAATC GCTGACGCTA TTCGTCACAA GGGTTATGGA AAAGATGTGC GTGAGGCAAT


 


31501
TGCACAGGGC TTCGAAAACT TAGATAAACA TTTAAGTAGT ATTGAAGAAG AACTGAAACA


 


31561
ACAAGAAAAG AAAAAGTCAT CGTCTATGGA TGATATTTTT AATTCTTTTG GTAAGAAGGA


 


31621
GTGATGATAA ATGGCAAAAG AGATTAGTAA TCTGATTACC TTTAACACCT ACAAATTTGA


 


31681
ACGAGGCGGC CTTTTGGTTG ATATGTTTAA CCAATTTAAT GCTCGTGTAG GAGATCAAGG


 


31741
AACGGAATTA GCCATCCAGT GGGAAACTAG TAAGACTGAA ACTAAAATTA ATTTAAAAGA


 


31801
ACGAGGATTA CATTTCTTTG GGACAGGTTC AGTTGGACAG TACCTTGAAA AATTAGAAGA


 


31861
TGGAACTGGC TTTAAGATGT CTGCAGATGC ATCTACCGTT GAATGGGAAG ATAAAGATGA


 


31921
AGCTGGTAGT TTAGACGATG GAATTACGGT TGTTAAGTTG CCAAAACAAT TTTTCCCTCA


 


31981
AAAAGGTATT TTCTTTGGTT ACTTTGGCCT AAAAGATAGA CAAGGTAATA TCTTTACTAG


 


32041
TGTTAATGTT TGGTTCCGTG TTCTTGGCGG TGTTCCAACG ATGGGGGCTG CTATTCCTTA


 


32101
CTTTGTTACT GAATTTGATG AAGTATTAGA GCGATGCAAT GGTAAGATTA TTGACGCTTT


 


32161
AGCAGAATTA CGTGAAAAGT ACCAGGCAGA AGTTAAGAAG AATGAGGATA TGTCTGCGGA


 


32221
AACAAGAGCA GCGTTGAGTA AACTTGCTGA TGCTGTTGGT GCAATTCAAG CGCAGATTGA


 


32281
TGCAGGTAAT GTAATTACAC GTAAAGAATA TAATAATCTT GCTAATCAGA TTGATAATCG


 


32341
CCTTAGTAAA ATGACACAGA ACATTGAGAG TTTTTCATCA CTCGATGACT TAAAAGCTCA


 


32401
ATATCCTAAT GGAAAAGATG GACTCTTTGT TACTAATGAC AATAATCATA AGTATCAATA


 


32461
CAAGAATGGA TCATGGGTAG ATGAAGGTAT ATGGACAGTT ACTACCTTTG ATCCAGAAAC


 


32521
GCGACGACGC CTTACATACC TTGATACATC TAATTCTATT TTGCAAAAAT CACTCAGTGA


 


32581
ATTGACTAAA GAAGTCGTTG ATATAAATTG GTTTTTAGGC GAAATTGATG CAAATACTGG


 


32641
TAAAATTACA CCTCACGAAA GTTTTAATCG TGCATACTCA TCACTCAAAG TAATTGGTAA


 


32701
ATCACATGCT TTTGAATTTT TGCTTAATAC TGATTATTTG CAATATATTA GTATTTTTGA


 


32761
ATTCAATTCA AATGGGGAGA TTATTAAGCA CGATAGTGTT GGCGATAAAG CGATATATAC


 


32821
ATTTGAAAAG GATACGACTG CGGTAAGGTT TCAGATTACT AGTACTGCTG CTACTATGAA


 


32881
GCAAAATGAT TTACGCGATA CTATTCAAAA TAGTGGGCTA AAAATTATCG ATCGAGGACA


 


32941
CCGTTCAGTA ATTAATGATA TTGATCATTT ATATATCCGT GAACATTTAA TGCCGATTGA


 


33001
AGCAATTGAA GGCTATACAA TTAACACTAA TGTTGATTAT GGTGAAACTG TTGATGTATC


 


33061
ACATCCAATA GTAACGTCTG CGTTTCAATA TATTAATCAA GTATGTAAGC CGGGCGATAT


 


33121
TTTCGTAATT AATAATCTAT CTGGAGGCTT TAACGCAATG GCTTGGGCCT TTATTGATAG


 


33181
CGAGAACCGG TTAATTCAAA AATCTGAAGT AAACTTATCT CAATCTCAGG TTACTTTATA


 


33241
TGCGCCAAGT AATGCTGCAA AATTAATTAT CAATAATATG GATAGTAACT GTACAGCATA


 


33301
TAGATATACT CCGGATAAAG AACATTTAGC TAAAATTAAT ATTGCTTTAA CTACCAATAT


 


33361
TCATCGAAAT ATCAAGTTAG CTTATCAAAC AGGAGAAACA GTCTCATTAA TTCCTGAAAA


 


33421
AGTAAGTAAT TACAAATATA TTATATTAGA TTGTAATTTT AATGATGCTT TCAGAATAAA


 


33481
GGGTTATGGT GGATTGAATC CACGATTATA TGGATTTATT TCTGAGCAAA ACACATTAAT


 


33541
GAATGTTGCG CCTGCAAATG TTAATGATCA ATTAACTGAT GTCTTTATCA AAACGCCAAA


 


33601
AGGCGCCAAA AAATTAGTAG TTAATTTCAA TCTTGATCAA CAAGTAAATC CCAAACAAAT


 


33661
TGCTGAACTT TATAAGCTTC CAGACATTAA TGTTATTACT GAGCAAAAAA TAAAAAAGAA


 


33721
TTTATTGAAT CAACAAACAG TCGATACATT AATTTCTAGT TATATGAATG GTATATTACT


 


33781
TGGTAAAGAT TTAGCCAATC ATCTTGCGGA TTTTGCTAAA AGCGGCGATA AAATGGTTCA


 


33841
TGTATCAACG TTTTATAAAG TTAACGATAC TCTTTTTATG TCTTATTATG CTAATACAAG


 


33901
ATCGGCTTAC GAAGATCCAA CCCAGCATAC AGCACGATTA GTTTATGCAC CGTTTGAGAA


 


33961
TCTAAATCAA CAAACGTACA TCGATGTAGC CGACATTGGA CAAGAATATA ATGGTCAAAA


 


34021
GATTGAAGCA ATATACGATT CTTTGCTTCT TAAAACAAGT GATGATTCAT ATATGATCTA


 


34081
TGCCTTTACT GCTAAGGTAG GTGGTAAGTT CTATATGCTT TACCGTCGTT TTGATCCTAA


 


34141
GACTAAGCTG TTGAGCGATA TTCATACAAT GAATTTTAAA GTTGGAGTAA TGACGTCGAC


 


34201
ATTTGATACA GTAAGCGTTC ATGATTTGTT AGCTAAGGCA GGAATTGACT ATGATTATGA


 


34261
AGATCGTGAT ATTTCCTTTG TCCAAAAATT AAGTCCACGG ATTGAAGATG GCGTAGTGCA


 


34321
ATACTATGCA GGTATCGGAA TCTTACATTT TTGCTTTGTA GTAAAATCAA GCGACTTAAT


 


34381
CAACTGGACT TTTGTCAGTA CTCCAGACTT TATGTACAAG CCAGAATTTG AACCTTCTGT


 


34441
ATATGTTAAG GGAGACAATG TTTATTATTT TTGTCGTCAG CGAGGGACAG AAGGTAATGC


 


34501
AGTTTTAGCT AAGTATAATA TCCCTAATGG TCAATGGTCG AATCCGATTC TTGTACCAGA


 


34561
TACTCAATCA CGCTATGACT TTTTCGAGAA TAATAGTCAG TTGTACCTTG TTCATTCACC


 


34621
GCTAGACCGT AATCATATTA GTTTGATGCA GATTGATCAG AATGTTCTGG AAAAGAGCTA


 


34681
TGAAGTAGCA ACAGCAGTAG TACAAGATTG CTTCTACCCA TTTACTCAAA ATATTGATGG


 


34741
TCAGATGTAT ATGAGCTTTA CACAAAGTAG GCAACATATT TGGCTAAATA AATTTAATCC


 


34801
CCACAGTCTG CTAGATAGTG ATGTAGCAAC TATTTTCAGT AATTTAATTG AGTAGCAAAA


 


34861
CATAGTCGCC TTATAAATAC ACAATACATA GAAAGTTGCA TTGCTTAAAT GGAAACTTAA


 


34921
TGGGCGGCTT ATAGACTTTT ATTATTTAAA AGAGTATACT GGAATTATTC TTTTTATTGG


 


34981
GAGGAAATAT AAAAGTGACA ACTAAAGTGA GAGATAAATC GCTTGATATT ATCCGGGGGA


 


35041
GCATCCTTCT AGTTGTCTTG GGTCATATAT CTGGGATACC TTTTGAGTTA AAAAAGTACA


 


35101
TATATTCTTT TCATATACCA CTGTTCTTTT TTGTTTCGGG ATATTTGTTC AACTTTGCTA


 


35161
AATACAGGTA TTTTTCTTAT AAAGAGTTTA TAAAATATAA AGCTAAAAAA TATATCCTAC


 


35221
CTTATTTCAG AATGGGGTTG ATATGTTTGC TTCTATTTGG CATAGTTTAT CCACTATTTG


 


35281
CTGAAGGGTT TAGTAAACAG TATATGCTTC AATCTACAAA ATATGTTTTA GGTTTACTAT


 


35341
ACTCGCGTGG AGGTCCTAAT TATATGGCCT GGAGTTCGCC ACTTTGGTTT CTAACAGCCT


 


35401
TATTTATTGC AGAGATTATT TTTTTTGTGG TTCTAAAATT TAATTTTAAA TATCCATTAA


 


35461
TAGTGTTTGG AATCTTAGCT ATATTGAGTT ATATTTACTC GATTACAATT AAAATTCCGT


 


35521
TACCGTGGAA TATTGATGTT GCAATGTTTG CAGTGTTATT CATGTACCTA GGCTTTATTA


 


35581
CACACAAATA TAATTTAACT AAGCACATTA ACTTACCGGT TTTCTTACTT TTGATTGTTA


 


35641
TTTTTGTATT ATCTGTAGCT TACAATAATG AGATTGACAT GAATTTAAGA AACTATGGCA


 


35701
ATGGATTTTT AACAATTATT AGTGGAACAA TAGGTACTGT TATATGCTTG CAAATTGCGC


 


35761
GATTGTTAAA AGAGAATAAG ATATTAGAGT TTTATGGTAA AAACACATTA TTTATCATGG


 


35821
GCTACACGTA TGCTGTGTTT AATTGTATTT TGGCTTTAAG CAGCCATTTT AGTACTGTAA


 


35881
AGAATGTTGT AGCTTCGTTT TTGATTCAAA TTATAATTTT AACTTTATTA ATAGTACTGA


 


35941
AGAATTTATT CAAACAAATA AAAAAGCCCA TTTATGCATA TACTAGGAAA ATTAACAATT


 


36001
AAAAAAACTA ATACGTCCTA CTCAGGGCGT TTTTATTTTA CCTTCAAAGG AGATGATCAA


 


36061
CATTGCCCTA TCATTTATTT ATGCTGCACC AAATGCAGAC ATTAGTTGAT GATAAACTAA


 


36121
TGTGGGCTTT TACAATCGTG ATGATTGTAG ATTTAATTAC GGGAATGATT AAACCGTATT


 


36181
ATGCAAAGAA AACAATTAAG AAAACTAATA GTTCAGTTGG AATCCCTGGG ATAATTAAAC


 


36241
ACACAGTAAT TTATTTAGTG GTAGTAATTG CTTATCCATA TCTTTATACG ATTGGAGCAA


 


36301
GCACGATGGC TACCACTTTT TTAATTGCTT GGATTTATCA ATATTTAATT TCAATTGTAG


 


36361
AAAATTGGAC AGAGATGGGG TGGTGGTTGC CTAAACCAAT CATGGATTTC TTTGAAGCCA


 


36421
AATTAGCTAA GGATCAAGAA GATTATGACC CATCTAAGTA CAATTTTCTT GGTAAATATA


 


36481
AAGGAGGTAA AAAGTAA.






Example 4. Selection of L. reuteri Promoters and Transcription Terminators
Global proteomics analysis of L. reuteri 3632 pellets and culture supernatants is performed to identify potential strong promoters and secretion signals, respectively. The proteins in the pellets and supernatants are ranked by their expression level, and the promoters and secretion signals from highly expressed and secreted proteins are identified by whole-genome sequencing (Example 2). The genetic elements are selected for use in expression cassettes that can be used to generate a genetically-modified bacterium which deliver the desired biomolecules to host animals in need thereof. Expression cassettes would comprise a suitable promoter, a heterologous coding sequence encoding a desired biomolecule, and a transcription terminator. The heterologous coding sequence could comprise a signal sequence for secretion, a cell-wall anchor sequence, and/or a detectable peptide tag.
Six suitable promoters are identified through analysis of the global proteomics data. Those promoter sequences are:








Xylulose-5-phosphate phosphoketolase promoter



(SEQ ID NO: 14)










  1
TTAAAGTATT AAAATAGATG TAAAATTTAT TTTTTTCAAA AGAAATTTTA ATTGTACACT



 


 61
GTTGGTATTG AACGGGGTTA AACAAAGGTA AATTAGCATT TCTGCGGATT AAGATAAATA


 


121
GAAAAATGTT AAAGAACACC TTAAAAAGAT TAATTTTTTA TAATTGGACC GTATCAATTT


 


181
GTAAAAAGGT TGACTTTTTG AAAAAAAAGT TTATCATTAA CATTGTAAAT TTAATGATTT


 


241
ACGTTATGTT GTTATAGAGC ACAGGACGTA TTGATTTATA TAGAAGGAGT GTTTATTAGA;


 








Elongation factor TU promoter



(SEQ ID NO: 15)










  1
ATGAATGGAC AGATGTTTTA ATCGCTAGAA TAGAAGGAAA GAAAGTCGCA ACAAATACGG



 


 61
TTTCTAGTAC GTGGCAGGAA CGACTAGGTA AGCAGATTGA CGAATTAATA GAAAAACATT


 


121
AGTCAAATAC ATTTACAAAT GAACAGATAG TTGATATTAT ATTTAAGAAT TCTTCTTCAG


 


181
AGCCTAAGAT TAAAGCTTTC AATTGGCGAA AAGAAGTTGT ACAATATGTA TAAAGGTATG


 


241
TCAGTCACCG AATCAGATGA TCTGGCATTA TACTTGTAAA TTATCAGGAG GTTTTCATTA;


 








Glyceraldehyde-3-phosphate dehydrogenase promoter



(SEQ ID NO: 16)










  1
ATCTCACGTG CGATCCATTA CACTAAGGGC GCGTCAACAA ATATTATACT ATCTTAAATA



 


 61
AGAATGAATT GCAAGCATTA TTTGAAAATT TTAATTAAAA TAACGCTTAC ATCAGAAAAA


 


121
TGTTGTGATT GAATAGACAA TTTTTTTGAA GATGGTATCA TAAGTATCGT AGGAGTTGTA


 


181
TTATTGCTTA GACCTTACCA CTGCGTCACT TACAATGGTT GAGAGTTGCG ATGCTGATGT


 


241
AATGTGATAA ACTAAGCAAG TACACTAATT ATGTTTTTTC CTAAAGGAGG AATTTGCAGT;


 








Glucose-6-phosphate dehydrogenase promoter



(SEQ ID NO: 17)










  1
TTGTTTAAGA TATCTTTCAA AGCTGCGGAA TTTTTCCCAG CTTTTTTAGT TAGTTTTGTT



 


 61
TTCATAAGCT ATAATTTTAA CCGATTCCAA ATTTCTTTTA AAAGTTTTTT TGATCTAGAC


 


121
CATTAATTGA TAAACGCTTA CCAAAGACTA ATCAACAAGC CATTTAGCGG TAGTGGTCCA


 


181
TTTTAACTTT CTAAGACATC TTCTCAGAAA ACGTTTCCTT TGATAGTGCA GATTGTGCTT


 


241
TAAGAGTATA TAATTGTCAC GGTATAAGAA TTTTCTGAAA TTTCAGAAGG AGTGAACATT;


 








L-lactate dehydrogenase promoter



(SEQ ID NO: 18)










  1
CTCCTCTATT ATTATTCCTG ATCAATTTTA AATTAATCTC CCTAGATAGG TATATTTTAG



 


 61
CACAGGTCAC CAACGTTCCA AAGTTTAATC TATGTTTAAA CTTTAATTTT CAAAAAAATG


 


121
CTATACTATG TTCACGATAC TTTAAGGAAA GGTGATTACA ATAGTGAGTC TCTTAATTGC


 


181
TATTCTTATC TGCTGGTTGC TATGGAAGAT TGGGGGTTTA ACGGTTAAGT TCATTGGTCT


 


241
AATCCTTCTT ATTCTATTAA TCGGGACATT AATTCATGTT TTACTTTGGC CAGCGATCCT


 


301
TTTAGCAGTT ATTATCTTAG GAGCAGGTTT ATTCACTAAC TAATTTATCT ATAAAATCTT


 


361
ATAGTAATTT TTCTGCGGAA TGTTATAATC ATTACTGTGA GAGAAATCTC AAATAATGTA


 


421
TACATAAGAT GAAAGGGAGA CTGTTTATT; 


and



 



tuf promoter








(SEQ ID NO: 19)










  1
ACAAATACGG TTTCTAGTAC GTGGCAGGAA CGACTAGGTA AGCAGATTGA CGAATTAATA



 


 61
GAAAAACATT AGTCAAATAC ATTTACAAAT GAACAGATAG TTGATATTAT ATTTAAGAAT


 


121
TCTTCTTCAG AGCCTAAGAT TAAAGCTTTC AATTGGCGAA AAGAAGTTGT ACAATATGTA


 


181
TAAAGGTATG TCAGTCACCG AATCAGATGA TCTGGCATTA TACTTGTAAA TTATCAGGAG


 


241
GTTTTCATTA.






The underlined regions of the above sequences denote the ribosomal binding sites.

Four suitable secretion signal peptides are also identified through analysis of the global proteomics data. Those secretion signal sequences can be derived from the following proteins:








C protein alpha-antigen precursor



(SEQ ID NO: 20)










   1
MVSKNNHQFY QQKHAERKQR WGIRKLSVGV ASVLLGTTFM LYGNHAVLAD TVTSPSDDVT



 


  61
RSTTTQGGNK DKVTEGTTEG TTSTPQTSGD STDKQANGQN VNQQVPTTDT EEATNHQDTP


 


 121
QGQDTTQNTT NVDKKDTEVT PANDATTPTT QKITAKFTTA KFTTAKFTAA KFKVLAARPV


 


 181
MKVAGTASLP ISNQDIKLDS QPMLTEIINK PTDNWVYNNL KWYQDTSTEK IKEILQNHTA


 


 241
NDESGRYYFA GVANYNEHYH AIYLLARSNN LNDNSLYVTI LHTGLGKNIQ EAVVAPGESK


 


 301
KVEYSGTTHT PIFTNYDGTS ASIDLDGIEK GDNIYGMVVG FAYGHNTGIK GDPASMGNGF


 


 361
VMTPIPTKMT TTIHYIDQAT GDEIAVPKSF EGVAYQKYTI TGEAPTIDGY TLKKSPETTG


 


 421
YISPYKVGES YDFRLDKHVV IKQTVIDAQG LVRVTAYYDG EVLNNTTRYL GNKLNVNDRM


 


 481
SFISHGKWYT YINQITSTND GIVYYYAKDG SEDKSEVRVH YIDVTGSKNS IFVPGDGEEV


 


 541
ATDKISGKLG ENYNYDVNLP TDYNLATNQA NTVNGTYTID HHDEYVYVVK KTSAELDPTV


 


 601
PAKTKVDNPT SLTADEKKTI EDKIVEANKD KFPEGTGVTV ANDGKATITY PDKSVDTIEG


 


 661
NQLVEEKTSA EKLDPTVPAK TKVDNPTSLT ADEKKTIEDK IVEANKDKFP EGTGVTVAND


 


 721
GKATITYPDK SVDTIEGNQL VEEKTSAEKL DPTVPAKTKV DNPTSLTADE KKTIEDKIVE


 


 781
ANKDKFPEGT GVTVANDGKA TITYPDKSVD TIEGNQLVEE KTSAEKLDPT VPAKTKVDNP


 


 841
TSLTADEKKT IEDKIVEANK DKFPEGTGVT VANDGKATIT YPDKSVDTIE GNQLVEEKTS


 


 901
AEKLDPTVPA KTKVDNPTSL TADEKKTIED KIVEANKDKF PEGTGVTVAN DGKATITYPD


 


 961
KSVDTIEGNQ LVEEKTSAEK LDPTVPAKTK VDNPTSLTAD EKKTIEDKIV EANKDKFPEG


 


1021
TGVTVANDGK ATITYPDKSV DTIEGNQLVE EKTSAEKLDP TVPAKTKVDN PTSLTADEKK


 


1081
TIEDKIVEAN KDKFPEGTGV TVANDGKATI TYPDKSVDTI EGNQLVEEKT SAEKLDPTVP


 


1141
AKTKVDNPTS LTADEKKTIE DKIVEANKDK FPEGTGVTVA NDGKATITYP DKSVDTIEGN


 


1201
QLVEEKTSAE KLDPTVPAKT KVDNPTSLTA DEKKTIEDKI VEANKDKFPE GTGVTVANDG


 


1261
KATITYPDKS VDTIEGNQLV EEKTSAEKLD PTVPAKTKVD NPTSLTADEK KTIEDKIVEA


 


1321
NKDKFPEGTG VTVANDGKAT ITYPDKSVDT IEGNQLVEEK TSAEKLDPTV PAKTKVDNPT


 


1381
SLTADEKKTI EDKIVEANKD KFPEGTGVTV ANDGKATITY PDKSVDTIEG NQLVEEKTSA


 


1441
EKLDPTVPAK TKVDNPTSLT ADEKKTIEDK IVEANKDKFP EGTGVTVAND GKATITYPDK


 


1501
SVDTIEGNQL VEEKTSAEKL DPTVPAKTKV DNPTSLTADE KKTIEDKIVE ANKDKFPEGT


 


1561
GVTVANDGKA TITYPDKSVD TIEGNQLVEE KTSAEKLDPT VPAKTKVDNP TSLTADEKKT


 


1621
IEDKIVEANK DKFPEGTGVT VANDGKATIT YPDKSVDTIE GNQLVEEKTS AEKLDPTVPA


 


1681
KTKVDNPTSL TADEKKTIED KIVEANKDKF PEGTGVTVAN DGKATITYPD KSVDTIEGNQ


 


1741
LVEEKTSAEK LDPTVPAKTK VDNPTSLTAD EKKTIEDKIV EANKDKFPEG TGVTVANDGK


 


1801
ATITYPDKSV DTIEGNQLVE EKTSAEKLDP TVPAKTKVDN PTSLTADEKK TIEDKIVEAN


 


1861
KDKFPEGTGV TVANDGKATI TYPDKSVDTI EGNQLVEEKT SAEKLDPTVP AKTKVDNPTS


 


1921
LTADEKKTIE DKIVEANKDK FPEGTGVTVA NDGKATITYP DKSVDTIEGN QLVEEKTSAE


 


1981
KLDPTVPAKT KVDNPTSLTA DEKKTIEDKI VEANKDKFPE GTGVTVANDG KATITYPDKS


 


2041
VDTIEGNQLV EEKTSAEKLD PTVPAKTKVD NPTSLTADEK KTIEDKIVEA NKDKFPEGTG


 


2101
VTVANDGKAT ITYPDKSVDT IEGNQLVEEK TSAEKLDPTV PAKTKVDNPT SLTADEKKTI


 


2161
EDKIVEANKD KFPEGTGVTV ANDGKATITY PDKSVDTIEG NQLVEEKTSA EKLDPTVPAK


 


2221
TKVDDPTKLT NDEKKEVEDN IRDHNTGLPE GTKIAVGDNG DTTITYPDKS VDTIEGNQLV


 


2281
EEKTSAEKLD PTVPAKTKVD DPTKLTNDEK KEVEDNIRDH NTGLPEGTKI AVGDNGDTTI


 


2341
TYPDNSVDTI PGDKVVEGKS DAAKNEPKVP GDKVKVDDPN KLTEDEKSEV VKAVEDANKD


 


2401
ENGKSTLPEG SKVTVGDNGD VTVTYPDGSK DTIPGDKVVE GKGTEGQTDA DKNEPKVPGD


 


2461
KVKVDDPNKL TEDEKSEVVK AVEDANKDEN GKSTLPEGSK VTVGDNGDVT VTYPDGSKDT


 


2521
IPGDKVVEGK GTEGQTDADK NEPKVPGDKV KVDDPNKLTE DEKSEVVKAV EDANKDENGK


 


2581
STLPEGSKVT VGDNGDVTVT YPDGSKDTIP GDKVVEGRGT EGQTDADKNE PKVPGDKVKV


 


2641
DDPTKLTEDE KSDVEQAIKD ANKDENGKST LPEGSKVTVG DNDDVTVTYP DGSKDTIPGD


 


2701
KVVEGKGTEG QTDADKNEPK VPGDKVKVDD PNKLMEDEKS DVEQAIKDAN KDENGKSTLP


 


2761
EGSKVTVSDN GDVTITYPDG SKDTIPGDQV IEGKSDADKN TPNVPGGDKV KVDDPTKLTD


 


2821
NEKNAVKDKV DEANSNLPDG TKVTVGDDGT TTITYPDGST NTISGHDLVT GKTDADKYPL


 


2881
NPGQAVNVVD PNHLTQAEQD QVKEAIQTTN PTAPIATITV DTAGNVQVTF ADGSTTTLQA


 


2941
NLHKHVTEAT TGSAIKPGVG TNGGQTKGAT STNQTATKQQ AQQHLPQTGD QPATWAMLSG


 


3001
LGVAFLGLLG LKKKRED; 


 








Arabinogalactan endo-1, 4-beta-galactosidase  precursor



(SEQ ID NO: 21)










   1
MEIKKHFKLY KDGKKWCCAA IATTVLGIGL AIGSPSVLAD ADTITSTSDA NNSLVKNDNT



 


  61
SDTDSNSEST FTDTNKNSTN EKEINENKNI DSSQQINQEQ TKSNNSEEQT TPVNVKAENT


 


 121
DIKDSIPEKS TPNSFKEING STYYYGENGD LYRNQFYNNW GRTYYFQANG ARLDNGFYNN


 


 181
WGRTYYFGSD GARWDNRFYN NWGRTYYFQN DGSRLDNSFY NNWGRTYYFG VDGARWDNRY


 


 241
MVKWGRAYYF GNDGALLQNQ LKSINGINYW INNEGIIPLK NQFLTANENQ LFYFDGNGSL


 


 301
VVNKFYHNWG HTYYFGSDGA RYTDQFLNRD GKVYYFDNQG IMYQDQYYKN WGHTYYFGSD


 


 361
GARYTDQFLN RDGKVYYFDN QGIMYQDQYY KNWGHTYYFG SDGARYTDQF LNRDGKVYYF


 


 421
DNQGIMYQDQ YYKNWGHTYY FGSDGARYTD QFLNRDGKVY YFDNQGIMVT NQVRVIDGKG


 


 481
YEFNDNGEAT ETSDMGQTRD TVAKEVAQAL TNQGIKGVKY DWRNTNNDYQ ELALHDIAQE


 


 541
VAQGDTNPDK NVIEKKLQAN NLLSGKVLVV YSTDFTNDDP QKITNTFMNS YDFTNADNSV


 


 601
LGVGADLNKN KLVIILFKPG EKAEQPQATS TISASISDIF KKAGVNVDVD NGLTKGSVVN


 


 661
SADLGNALTN GTAELLKGDK GTIISQEVLK AIFAAFAGNT SAVEGTKNYY NGNDAYHYEF


 


 721
WLEGQSADDK LNNFLALNKG AKYGDQLKVN YTATLVFGQE TGTNSNESKV PASERTDEQL


 


 781
DLAYKTGTDT GLRYDSVKVE KIPGMTDDMV RGVDVSSYQA LINAGVKFYD FNGQESNLFK


 


 841
ILKDSGVNWV RLRVWNDPYN AQGQPYAGGD NNEENLIKMA KEASDNGLKL LIDFQYSDFW


 


 901
TDPAQQILPK AWRNLSHGEM SQEVYLYTSK ILNDLQKAGA SVKMVQIGNE ITNGAFGLYT


 


 961
GRNGGGNWAS LWETSDGDQV AKYIQAGSSA VRRIDPTIKV AIQLETPEIN KYRGIMNVLK


 


1021
KNNVDYDYLG TSYYPFWSTT QGNGWYDNVD LGYGANTPVN LEAIEKMAWN EFGKRTVILE


 


1081
SGWLNNTNDA DGTHNSVGEN NETTNIDRYS ADPQGQVDEI EDMYNAIIAQ KGLGAFYWEP


 


1141
AWIPVKAGWN NWQYNKLMSN IYGSGWASQY AKGYAPDSVL YYDGKEAWGG SSWDNISLFD


 


1201
DHGHPLQSLN VYNGMLNGYE SPKNVKSSLS TQLVKIWNET DVIPNDGLTE GTKLSTDLFG


 


1261
TTQLSGNDGQ SIGNAELTKL AGRLKDGISS KVYTAANGAR YHYIYWLEGG NNKVNTFVSA


 


1321
NKDAKYGQPL IANYSATVVV DSEPGTQVAT SPLQIKISQV WNTVNNEEIK IDNPLKQGDL


 


1381
ITDKSDNAFS GILNSKDIKE ALTGEKGKDV SESTVNDVKS LLPKEVKGSK TYTTADGNQY


 


1441
YYDFWLASVE TSNVNYGEPI IVNYTASLKW LG;


 








Chromosome segregation protein



(SEQ ID NO: 22)










   1
MEKTMKKKAL VATTAVAGIT LVGEVTTVHA ADNVQQPVNE QNVNQSSQEE KQAAQNLQNA



 


  61
QSDVNTATEA NSNAQDNLAS ANNNLSNAKK AVSDQAAKVA DATKAQSDAS TKVDNDNKVV


 


 121
ADAQQKADQA TPANIENAKQ AIEGQNKVID QDNENIKYSN TDQDKAQNTL NNAQSNEDKA


 


 181
NATLSNKKSS QASAQNNVKQ AEDALNGTHL VEAQNAFNQA QSNVENAQSK YDQANNQLSD


 


 241
AQKKVTTNQN DLTAKNKALD NINNQVDTDQ NNVNSNQATA DSASSATQVA QNAVDQTKQS


 


 301
LDKVIEELNG FSENTIKVPA GAQEAYEAFI DAVDNNADQS QLDSLAKKMY DTLHQGQGTN


 


 361
GINHFNSSKY DQNQLVDVDH LTTDQLNELT QFAADMINSA RKAWGSDKNA GTLIPTQGVS


 


 421
EMAQQIAKGY VSDNWHISQG HDVKRVTAAA GLIGLNDAGQ FYEDASEGYV HAWPWEKDSY


 


 481
TMDNLKEAVY DSILGMLFAD DNSGNGHMTD LLGLHVNRKE DHQYFGLSTN MCPGSYMGQL


 


 541
HFIIVENDPA YIKDPQTFNA KGGTTKIEYI DPKVQLNQQK DILTTTLSTQ QADLATKQDA


 


 601
LNKANQNLAN AKKQLSEDQD LQTVAQQNRD SAQKALNDAT AKVSNLQATV NSLSQDLNSA


 


 661
KATLDQAKKT LESYTADHKA KLDNYNNAKA ALDDANKAVA EAQSAVDTAV NETKIAQNNL


 


 721
DQKKQAVTDA QNKLANDQEY LATLKQNLAD LQNAPQNLQK AKDQLAKDQI ALDNANKDLQ


 


 781
NQKDSLDELN KKLEDAQVKV NEAQSAANVT KATLDQAQAK LSDAEATWKE LHNDAHRYGN


 


 841
VVKVTPITME AGTSLPDPVI ENGFTVNTGT NQLFVSLAAI DSSNNNIPQG TKASWANRSK


 


 901
ALTDSQNAGS YSEDILITFP DNSTVTVPVD LTVTAKKITE DQKATEGGYH IVNGSVVDKQ


 


 961
NNLVSGWTVK NGQMVDPEGN VIKTTMSTAQ GVTIEKNNSK SGNTKTNMIQ TSLTIANNKA


 


1021
TTNKDNQLPQ TGNYNNNTKV LGLAGIALAS ALTMFGYKKR QHN;


 








D-gamma-glutamyl-meso-diaminopimelic acid endopeptidase Cwls precursor



(SEQ ID NO: 23)










   1
MKSTTKKILA SSLGVAGAMA MGTVTAKADT TVTVNAGDSL NGIAQKYNVS ADDIATANHL



 


  61
QNKELIFVGQ KLTIPTKDKN ETPANNAEKK DQASKNSQSL QDSVNKAMSY LGTPYVWGGN


 


 121
KPGGFDCSGL VQYCYGIPQR TTYEQQALGP HIHDNVLNAP YGALVFYGSD DAPYHVAISL


 


 181
GDGRIIQAPN ENETVKITDQ QYFPGNYYVV MH,


and



 








Chromosome Partition Protein Smc



(SEQ ID NO: 40)










   1
MNKANQKVAD DTTAVNNKQT DVNNAAEAKK NADEALKNAN DAQTSAQKNK DAKQAIADEA



 


  61
SVALADANTA VKDAQAKVDA INDKLANFNT ITLPAGYKDD LIAYYNYFGN SNYNQDEANN


 


 121
LAQDLLKYRD QAMSQNKFKD NLSDDRVVDI DNLNSTDRAE LSQFVASLIN QVRTQMGTNL


 


 181
VISSPAADDY AEQVSQNYNK DNWNSADNGK HDQSALNNAT DQLNISWNGE NMGLDQSIFT


 


 241
TDYTVLTDGT KLPTGNKQTI NDLKHLIYDD FISMMFDDAD SAWGHATNFA GIDNFAAEKQ


 


 301
AVGFSLDKFY NTHYDLVEAN QKVEENSYTL PSINALTQKL ADAKDDLSIK QTDQASKQKA


 


 361
NDDAQNALSS ANQVLVAAQN DVKDKTATAQ EANDNLTTAQ NDLATLQNQL SADQANQKQA


 


 421
QTTFDSFDAD LATKQANLQK ATDSLKAEQG RLAIAQADLD NANKALSDAN NNLAQKKQVV


 


 481
ENDNETLKVD NDKLVQLQNN LSDLQNAPKL LAAAKEQVAT AQKALADAQE AYNVANDKLT


 


 541
SLKQTAAGTT TNVSKAQQAL AEAKNNEDAA KEVLDQAQQA LTELRQKEAL AKQVAEEQAK


 


 601
LAAEKEAKDN GYHIENNQVV DAKGNSVNGW TVKGNQIVSP TNATVDPAVS VTTNVNVDSK


 


 661
GQVQPQTSVT ANSVKTVAAT ESANPVATTT VQTREQYKQQ LKSNNQLPQT GNNDSAVLSL


 


 721
AGVALAAMLS LFGIKKREY.






A person of skill in the art would recognize that, because of the redundancy of the genetic code, multiple nucleic acid sequences could encode the above peptides.

Example 5. Expression Cassettes
Based on the information provided in the previous examples, expression cassettes that produce high levels of secreted target biomolecules can be designed. For example, one of these expression cassettes (CwlS_C2; SEQ ID NO: 23) has evolved to deliver endopeptidase (cell wall hydrolase) to the cell wall. That endopeptidase plays an important role in cell division and separation. Replacing the endopeptidase with a heterologous coding region would result in the secretion of desired target biomolecules, such as anti-infective biomolecules that target pathogenic bacteria. A specific example is that the novel mersacidins disclosed herein could be expressed at high levels by exchanging the native mersacidin promoter with a strong promoter (Example 4; SEQ ID Nos: 14-19) and optionally a strong secretion signal (Example 4; SEQ ID Nos: 20-23).
Expression vectors are designed and synthesized with different combinations of some or all of the following components: an origin of replication for replication in L. reuteri (e.g. from the high copy number plasmid; SEQ ID NO: 8), an origin of replication for replication in E. coli, a drug resistance marker for selection, a strong promoter for expression in Lactobacillus, a signal sequence for secretion, a heterologous coding region encoding a desired biomolecule, an expressed peptide tag for detection, a cell-wall anchor for secretion, and terminators for transcription termination.
Alternatively, a heterologous coding region encoding a desired biomolecule could be integrated into the chromosome of the genetically-modified microorganism. Chromosomal integration of the expression cassette (a strong promoter for expression in Lactobacillus, a signal sequence for secretion, a heterologous coding region encoding a desired biomolecule, terminators for transcription termination, and optionally an expressed peptide tag for detection and a cell-wall anchor for secretion) is accomplished with a suicide vector. The suicide vector comprises an origin of replication for replication in E. coli, a drug resistance marker for selection, and an expression cassette flanked by nucleic acids homologous to a specific region of the chromosome. An example of such a suicide vector is presented in FIG. 3A.
For chromosomal integration, L. reuteri genes may be interrupted by the insertion of the expression cassette. Maps of these chromosomal loci are presented in FIG. 3B. Two genes which may be interrupted are Transposase and Uracil phosphoribosyl (UPP) transferase. Chromosomal integration may also be accomplished within the pyrE gene locus, and pyrE deletion and subsequent re-insertion may serve as a selection marker for integration (Sakaguchi et al., Bioscience of Microbiota, Food and Health, 32: 59-68 (2013)). A modified pyrE gene may also be used for biocontainment of recombinant bacteria, as the recombinant bacteria cannot survive in an environment absent metabolic supplements.
Example 6. Enzymatic Genes for Inclusion in Expression Cassettes
An expression cassette would comprise a heterologous coding region encoding a desired biomolecule. The desired biomolecule may be a biomolecule with anti-infective activity. The anti-infective activity could be lysis of pathogenic bacteria by a lytic enzyme, for example from a bacteriophage, with specificity to a certain genus of pathogenic bacteria.
Lytic enzymes may include PlyCM, a lytic enzyme targeting Clostridium perfringens, encoded by a sequence of:








(SEQ ID NO: 24)










   1
ATGGAAAGCC GTAATAACAA TAACCTGAAG GGCATCGATG TGAGCAACTG GAAGGGCAAC



 


  61
ATCAATTTTC AAAGCGTCAA AAATGACGGT GTTGAAGTTG TTTACATTAA GGCAACCGAA


 


 121
GGCAACTACT TCAAAGACAA ATATGCTAAG CAAAACTACG AGCGCGCTAA AGAACAGGGT


 


 181
CTGCGTGTGG GCTTCTACCA CTTTTTCCGC GCAAACAAAG GTGCCAAAGA TCAGGCGAAC


 


 241
TTCTTCGTGA ATTACCTGAA CGAAATCGGT GCGGTCAATT ATGACTGTAA ACTGGCACTG


 


 301
GACATCGAGA CTACCGAAGG CGTCGGTGCG CGTGACCTGA CCTCTATGTG CATCGAGTTC


 


 361
CTGGAAGAGG TGAAGCGTAT TACGGGTAAG GAAGTTGTCG TGTACACCTA TACCAGCTTC


 


 421
GCGAACAATA ATCTGGATTC CCGTCTGTCT AGCTATCCGG TGTGGATTGC GCACTATGGC


 


 481
GTCAACACCC CGGGTGCGAA CAATATCTGG AGCGAGTGGG TGGGTTTCCA GTACAGCGAG


 


 541
AATGGCTCCG TCGCCGGTGT CAGCGGTGGC TGCGATATGA ACGAATTTAC CAATGGTATC


 


 601
TTTATTGACT CGAACAATTT CACGTTGGAC AATGCAACGA CCAAAAATGT TAGCATTAAG


 


 661
CTGAACATTC GCGCCAAGGG TACGACCAAC AGCAAAGTTA TTGGTAGCAT TCCGGCGAAC


 


 721
GAAAAGTTTA AGATCAAATG GGTTGATGAA GATTACCTGG GTTGGTATTA CGTTGAGTAT


 


 781
AACGGTATCG TGGGTTACGT TAACGCCGAT TACGTCGAGA AACTGCAAAT GGCGACCACG


 


 841
CATAATGTTA GCACCTTTCT GAATGTACGC GAGGAGGGTT CCTTGAATAG CCGTATTGTG


 


 901
GACAAGATCA ACACTGGCGA CATCTTTCGT ATTGACTGGG TTGATAGCGA TTTCATTGGT


 


 961
TGGTATCGTG TGACGACGAA AAACGGCAAG GTCGGCTTTG TTAATGCAGA GTTTGTGAAA


 


1021
AAGTTGTAA;






PlySS2, a lytic enzyme targeting Streptococcus suis and Staphylococcus aureus, encoded by a sequence of:









(SEQ ID NO: 25)










  1
ATGACAACAG TAAATGAAGC ATTAAATAAT GTAAGAGCTC AGGTTGGGTC CGGTGCGTCT



 


 61
GTTGGCAACG GCGAATGCTA CGCTTTGGCT AGTTGGTACG AGCGCATGAT TAGTCCGGAT


 


121
GCAACTGTCG GACTTGGCGC TGGTGTGGGC TGGGTCAGCG GTGCAATCGG CGATACAATC


 


181
TCTGCCAAAA ACATCGGCTC ATCATACAAC TGGCAAGCTA ACGGCTGGAC AGTTTCCACA


 


241
TCTGGTCCAT TTAAAGCAGG TCAGATTGTG ACGCTTGGGG CAACACCAGG AAACCCTTAC


 


301
GGACATGTGG TAATCGTCGA AGCAGTGGAC GGCGATAGAT TGACTATTTT GGAGCAAAAC


 


361
TACGGCGGGA AACGTTATCC CGTCCGTAAT TATTACAGCG CTGCAAGCTA TCGTCAACAG


 


421
GTCGTGCATT ACATCACACC GCCTGGCACG GTCGCACAGT CAGCACCCAA CCTTGCAGGC


 


481
TCTCGTTCCT ATCGCGAGAC GGGCACTATG ACTGTCACGG TCGATGCTCT CAATGTTCGC


 


541
AGGGCGCCAA ATACTTCAGG CGAGATTGTA GCAGTATACA AGCGTGGTGA ATCATTTGAC


 


601
TATGATACTG TCATCATCGA TGTCAATGGC TATGTCTGGG TGTCTTACAT AGGCGGCAGC


 


661
GGCAAACGTA ACTACGTTGC GACGGGCGCT ACCAAAGACG GTAAGCGTTT CGGCAATGCT


 


721
TGGGGTACAT TTAAATAA;






and CP025C, a lytic enzyme targeting Clostridium perfringens, encoded by a sequence of:









(SEQ ID NO: 26)










1
ATGTCGAAGA TTTTTGGTTT AGATGCGGGT CATTGTACGA GCGGCGCAGA TACGGGTGCG



 


61
CAGGGCAATG GTTACAAAGA ACAAGACTTG ACCCGTCAAGT TGTTACCTA TCTGAGCGAA


 


121
TACTTGGAGA AAGAGGGCCA CACTACCAAG TACTGCCATT GCAATAGCGC GAGCACGGTT


 


181
AACGAATCCC TGCGCTATCG TGTGAACAAA GCCAACTCCA TCGGTGTCGA CTACTTCGTG


 


241
AGCATCCACC TGAACGCCGG TGGCGGCGTT GGTACCGAAA CGTACATCTG CGCGCGTGGC


 


301
GGCGAGGCCG AGCGCGTGGC GAAACGCGTC AATTCTAAAC TGGTGCAGTA CGGTTATCGT


 


361
GACCGTGGTG TCAAGGTTGG TAATCTGTAT GTGATTAAGA ACACCAATGC ACCGGCTATC


 


421
CTGGTTGAGA TCTGTTTCAT TGACAGCAGC AGCGATGTGG CAAAGTTTAA CGCGAAGGCA


 


481
ATCGCGAAAG CGATTGCTGA GGGTCTGCTG GATAAAACCA TTGGTGAAGT CGAGAATAAG


 


541
TAA.






Additionally, two of the three productive prophages found in L. reuteri strain 3632 contain putative lytic enzymes. These lytic enzymes may be used to target pathogenic bacteria or to control environmental spreading of genetically-modified Lactobacillus. These two lytic enzymes are: an N-acetylmuramoyl-L-alanine amidase sle1 precursor from prophage locus 1 with a nucleic acid sequence of:








(SEQ ID NO: 27)










   1
ATGCGTAATC AATTCATCGA TGTTTCAAGT TATCAACCAG ATACTGTTGC CTTTTTCCAA



 


  61
GCTGCTAAAG CTCAGGGTGC ATTAGGGGTC GTTGTTAAGT TAACGGAAGG GTCCGAAGAT


 


 121
GGTTCGGCTT ATGTTAATCC ACGTGCGGCC GCTCAAATTC GTAATGCCTT AGCGGTTGGC


 


 181
TTGCGCGTTT CCTGTTACCA CTTTGCTCGT TATACATCAG TGACTGATGC ACAAAATGAA


 


 241
GCTCGATTCT TCGTTAAAAT CGCTAAGCAA TTTGGTATGT ATGACGATAC TTTGATGATT


 


 301
GATGATGCGG AAGTTCATTC AACTGCAGAT TATCAATCAG TATCCTTAGC CTTTCTTCAA


 


 361
GAAGTAGAAG CTCTTGGTTA CAAGAATACT GGGATTTACT CCATGAAGTC CTTCTTCACT


 


 421
GGCGGTATTC TTAATTCACA TGGCTTTGAT TCCCGGAAGA TTTGGATTGC TGGCTATGGT


 


 481
GTGACTGAAC TGGGGATTGA TAATGCAAGT GCTTGGCAAT ATTCTGATCA TAGCATCATG


 


 541
GGAATTGATA CTAGTTATGA CTTTGACGGT GCCTTTACGA CTGGTTTAGT ATCAGGCAAT


 


 601
GTTCCGCAAG CTGTTATTCC AGCACCACAG CCGGTTCAAC ATATTGGTCA CCCAGCTACT


 


 661
GGAACCTACA TTGTTCAGCC GGGCGATACA TTGAGTGGAA TTGCAGAAAA ATACGGGACT


 


 721
ACTTATCAGA ACCTAGCAGC AATCAATGGT ATTGGTAATC CAAACCAGAT CAATGTCGGC


 


 781
CAAGTCCTCA AAGTCACCGG AAAAGTATCA AACGAAAATA CTTACTTTGT TCAATCAGGC


 


 841
GATACGTTAT CCGGAATTGC CACCAAATTC GGCACCACTG TCTCAGACCT CGTAAGCCGT


 


 901
AATCACATTA CTAACCCGAA TGTGATCTAC GTTGGGCAAA AACTCTACTT AGCCGGCAAC


 


 961
GGACAATCCA ATGCTTATAC TGTCCAAGCA GGGGACACAC TAAGCGGAAT TGCGGCTAAG


 


1021
TTTGGCAAGA CCTGGCAAGC ATTAGCTCAA AAGAATGGCA TCGCAAATCC TAATATGATT


 


1081
TTCATTGGTC AAACAATTCA GATTTAA,






and a Peptidase family M23 from prophage locus 6 with a nucleic acid sequence of:









(SEQ ID NO: 28)










1
GTGTACCGAA TTATTGGTTA TAATGAACCA ACAGATAAAG CAGGATTTAT TGTACTGGAT



 


61
CCCCGAGTTA ATCGTCATAT TAGTTCGGGA AAACTCACGC TTAAAGAATC TAATATTGAT


 


121
GATTTGACTA TTACGGTTAA TCAAGCAAGT CCATTATGGG ACAACGTAAG GCCTTATCAT


 


181
ACTCATGTTA ACGTTTATGA TGATAATGAA CTTATTTTTC GTGGACGAGC TATCAAACCT


 


241
AAAAAGTCGA TGGAAGAAAG CGGACAATTC ATTCGTGAAT ATGTTTTTGA AGATATTGAA


 


301
GCATATCTCA TGGATAGCAC CCAAAGATTT TATGAAGGTG TTGGTCAAAC GCCCAAAGAA


 


361
TTTTTACAAA CTTTAATCGA TGTTCATAAT TCACAGGTTC CTGACTATAA AAAGTTTCAA


 


421
GTCCGGAATG TAAATGTCAC TAATAATAAG GATGACCAAT ATCGACAAAT TGATTATCCC


 


481
AAAACTAGCG ATGCTATTAA TGATAAATTA GTTAAATCTC TTGGTGGTTA TATTGTGACT


 


541
ACTTACAACG CTAACGGAAT AAACTACATT GACTACTTAA CGGATATTGG GGTTGATCAT


 


601
AAAGATGATA CTCCTATTCA GTTAGCTAAA AATATGAAGT CTGCAAGTAT GCAAATTGAT


 


661
CCTACTAAGG TGATTACAAG ACTGATTCCA CTGGGAAAGA CACTAGAACC ATCAAAAGTT


 


721
GATGTAAGTG ATGATGATGG AGAGGGCGGT TCTGGATCAT TAGATAGCCC TGAAGAATTT


 


781
TGTAAATCAG AAATTAATGC TACTTGGGGT AGTGATATTA ATAATATGAA ACAAGATTTT


 


841
GCCGCTCGTT CTTCGAGAGT TCGGGCTTGG GGAGTGGACG TTAATCGTTT ATATGATGTG


 


901
GTGAAAAATG CTGGAGTAAG TCCTGAATGG TTCTTTGCTT ATGAACTTCA AGAACAAGGA


 


961
ACTTACTATG GATGGCTTAA CCATACTTAT CGACACGGTG ATGCGTATAG TGATGCGCAA


 


1021
TCTGTTTGTG AGTGGATTAA AAATTGTTCA AATAGTAATT CCATTAATCC AGCATGGAGC


 


1081
GCACCGGAAG GATCAATGGC GCCGAATCAA GCATTAGCGG ATAAATGGAA TCAAGAGTTT


 


1141
GGAAAAGGTA CTATTGGCCG CGTTTATTTA CAAGGGACTG CCGCTGCTGT TTGGGATTTA


 


1201
GCTGGTCAAA CGCCTAATCC AGCTATTGGA AAGCCAATTA GTGGATGCAT TTCTTGTATT


 


1261
AAACGTTGGG GTGGTCATTC TAATGCAGCT GGTGGTACAT GGGGATGGCC TTTTCCTGAT


 


1321
GTTGGGGAAG GTCATTTTTC TCAAGTTCAG AGTTTCGGAA ATGATGGCGG ATATCGTCAA


 


1381
AATAGTTATC ACGATGGTGT GGATTTTGGA TCAATAGATC ATCCTGGTAG AGAAGTGCAT


 


1441
TGTATTCATG GTGGAACGGT AACTATCAAA TCAGCTATGG GTGGCTTAGG TAATTTTGTG


 


1501
GTTATTCATA CGCCGGAAGG ATTCAATATC GTTTATCAAG AAGCTTTTAG TTCTCCCTCT


 


1561
AATATTATTG TTAGTGTTGG GCAAAAAGTA AAAACTGGTG ATGTAATTGG ATATCGTGAT


 


1621
ACAGACCATG TTCATATTGG CGTAACTAAG CAAGATTTTT ATCAAGCAGT TCGAAATTCT


 


1681
TTTTCTCCTG CAGGTGGTTG GCTAGATCCA GTAAAACTAA TTAAAGAAGG TGGCGATGGG


 


1741
TCTAAACCAC AAGAAGGAAA GAAAGATCAA ACTGTTGATA ATAGTAATGC TGCACGTCCT


 


1801
AAATTAACCA TTACTACTGT CAATAACGGT AGAGACTATA TTGATATTCC TGATTTACAA


 


1861
AAAGAATTCG GTATTATTGA GGGAACTGTT GAATTTGATA ATGTAGATGA TCCGAATGTT


 


1921
TTAATGCAAC AAGCTCAAAC ATGGATAAAG GCTCAAAGAA TACCTCAAAG TTGGGAAGTT


 


1981
ACAGCTTTAG AATTACATAT GACAAACTTC AAATCTTTTA AGGTTGCTGA TAGGTACATG


 


2041
TTTATTAATC CAAATGTTGC AAAACCCCAA TTATTACGAA TTACTCAAAA AGAAATTGAT


 


2101
TTACTAAAGC CCCATGCGTC TTCATTAACG ATTGGTGATA AGACGATGGG GCTTACTGAT


 


2161
TATCAGTTAG AAAATCAAGT CAATTTTCAA CAATTTAAGG AAATTCGAGT GATGGTTAAT


 


2221
CAGGTTGTCC AAACCCAAGA GCAATCTGCT AATAACAATA ATAAGGTTAT GCAAAATTTT


 


2281
GCTAGTAGTG CTGATCTTGC ACAAATGAGA CAGGATCTAA GAAATCTTCA AGATGATAAC


 


2341
GATCGTGCTC GCAAAGGAAT GGTTTCCTTA GAAGAATTCA ATAAACTAAA GGAACAAGTA


 


2401
GAAAAACTAA CAACAGGAGG CGATGATAAT GGCAAGTGA.






Example 7. Single Chain Antibodies and their Activity
An expression cassette would comprise a heterologous coding region encoding a desired biomolecule. The desired biomolecule may be a biomolecule with anti-infective activity. The anti-infective activity could be inhibition or neutralization of toxins produced by pathogens. The inhibition or neutralization could be accomplished with single chain antibodies. Lactobacillus has been described as an expression system for single chain antibodies directed against host attachment factors. WO2012/019054. Toxins to be targeted by single chain antibodies include C. perfringens alpha toxin and NetB.
Camelid heavy-chain only (VHH) antibodies against C. perfringens alpha toxin and NetB are prepared by QVQ Holding BV (Utrecht, NL). Briefly, two llama calves each are immunized with either recombinant alpha toxin or NetB variant W262A. Neither of these immunogens are haemolytic. The immunized llamas are boosted twice with toxin peptides. On days 44 and 72 after the primary immunization, blood samples are taken and RNA isolated for phage library construction. Phage libraries are screened for binding activity towards each of the two toxins. The candidate antibodies are sequenced and further screened in bioassays as described below.
Alpha toxin causes membrane damage to a variety of erythrocytes and cultured cells. It is preferentially active towards phosphatidylcholine (PC or lecithin) and sphingomyelin (SM), two major components of the outer leaflet of eukaryotic cell membranes. The N-terminal domain possesses full activity towards phosphatidylcholine but lacks the sphingomyelinase activity and is not haemolytic or cytotoxic. The C-terminal domain is devoid of enzymatic activity, but interaction between the N- and C-terminal domain is essential to confer sphingomyelinase activity, haemolytic activity and cytotoxicity to the toxin. Although alpha toxin is a potent haemolysin, the lysis of erythrocytes is only seen after intravenous administration of toxin in experimental animals or in cases of clostridial septicaemia.
The inhibitory capacity of the VHH antibodies directed towards alpha toxin on the alpha toxin lecithinase activity is determined by measuring its effect on egg yolk lipoproteins. Fresh egg yolk is centrifuged (10,000×g for 20 min at 4° C.) and diluted 1:10 in PBS. The ability of the VHHs to neutralize the alpha toxin activity is assessed by pre-incubating a two-fold dilution series of the VHHs (two wells per dilution, 5 μM starting concentration) with a constant amount of alpha toxin (either 5 μg/ml recombinant alpha toxin or 3.33×10−4 U/μl alpha toxin from Sigma, P7633) for 30 minutes at 37° C. prior to the addition of 10% egg yolk emulsion. As a positive control, serum from calves immunized with the recombinant alpha toxin is used, starting from a 1:4 dilution. After incubation at 37° C. for 1 hour, the absorbance at 650 nm (A650) was determined. Alpha toxin activity is indicated by the development of turbidity which results in an increase in absorbance.
The control serum is able to neutralize the lecithinase activity of both the commercial and the recombinant alpha toxin. An eight-fold dilution of the antiserum (corresponding to 3.12% serum) is able to completely neutralize the alpha toxin lecithinase activity of the recombinant alpha toxin, whereas only the highest dilution of the antiserum (corresponding to 25% serum) is able to completely neutralize the lecithinase activity of the commercial alpha toxin. Considerable difference in inhibitory capacity is observed between five candidate VHH antibodies. VHH EAT-1F3 had no effect on the lecithinase activity of either of the alpha toxins. The neutralizing capacity of EAT-1A2 and EAT-1C8 is very similar and is the same for both the recombinant and commercial alpha toxin. The maximal inhibitory capacity is preserved until a 32-fold dilution (0.16 μM VHH) of the VHHs. However, both EAT-1A2 and EAT-1C8 are unable to completely neutralize the lecithinase activity, resulting in 40% to 50% residual lecithinase activity. Two other VHHs, EAT-1F2 and EAT-1G4 show a difference in neutralizing capacity towards the recombinant and the commercial alpha toxin. EAT-1F2 has a high neutralizing capacity towards the recombinant alpha toxin but is unable to completely neutralize the commercial alpha toxin, resulting in about 25% residual lecithinase activity. In contrast to EAT-1F2, EAT-1G4 neutralizes 100% of the lecithinase activity of the commercial alpha toxin, but is less capable of neutralizing the recombinant alpha toxin.
Neutralization of the alpha toxin haemolytic activity by the VHH antibodies directed towards alpha toxin is determined by measuring its effect on sheep erythrocytes. Similar to the inhibition of the alpha toxin lecithinase activity, the ability to neutralize the haemolytic activity is assessed by pre-incubating a two-fold dilution series of the VHH antibodies (two wells per dilution, 5 μM starting concentration) with a constant amount of alpha toxin (6.25×10−5 U/μl alpha toxin from Sigma, P7633) for 30 minutes at 37° C. prior to the addition of 1% sheep erythrocytes. As a positive control, serum from calves immunized with the recombinant alpha toxin is used, starting from a 1:4 dilution. After incubation at 37° C. for 1 hour, the plates are centrifuged to pellet intact red blood cells. The supernatant is transferred to a new 96 well plate and the A550 is determined. Alpha toxin activity is indicated by the increase in absorbance due to release of haemoglobin from the erythrocytes.
The inhibitory capacity of the VHH antibodies towards the alpha toxin haemolytic activity is determined using the commercial alpha toxin only, as the recombinant alpha toxin shows no haemolytic activity. Up to a 16-fold dilution of the control serum (corresponding to 1.56% serum) completely inhibits the alpha toxin haemolysis. To the contrary, none of the candidate VHHs has an effect on the haemolytic activity of alpha toxin. Because the control serum contains polyclonal antibodies, whereas the VHHs are monoclonal, the combined effect of all 5 VHHs towards alpha toxin is determined (1 μM of each VHH in the highest dilution, corresponding to 5 μM VHHs in total). Combining the VHHs has no effect on the alpha toxin haemolysis.
Based on the above results, EAT-1F2 and EAT-1G4 are selected for further characterization. The peptide sequence of EAT-1F2 is:







(SEQ ID NO: 29)


EVQLVESGGGLVQAGGSLRLSCAGSGRTGSLYSMGWFRQAPGKEREFVA


 


AITWRPSSTYYADSVKGRFTISRDDAKNTVYLQMNSLKPEDTAVYFCAA


 


RPRGGLSPTPQAYDYWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHH


 


HHHHGAA,






and the peptide sequence of EAT-1G4 is:








(SEQ ID NO: 30)


EVQLVESGGGLVQPGGSLRLSCAASGSIATINDMGWFRQAPGKQRDWVAT


 


IVSDGSTAYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCSARRH


 


YGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA.






An R27H mutant of EAT-1F2 (SEQ ID NO: 49) may also be generated to improve protease resistance of the antibody.
A person of skill in the art would recognize that, because of the redundancy of the genetic code, multiple nucleic acid sequences could encode the above peptides. However, an exemplary sequence encoding EAT-1F2 could be:








(SEQ ID NO: 31)










  1
GAGGTGCAGC TCGTGGAAAG TGGCGGAGGT CTTGTTCAGG CTGGGGGATC GCTCCGTCTG



 


 61
AGCTGTGCGG GGTCTGGCAG AACAGGTAGT CTCTATTCCA TGGGTTGGTT TCGGCAGGCC


 


121
CCGGGTAAGG AGCGGGAGTT CGTTGCAGCG ATTACGTGGA GGCCCAGCTC TACCTACTAC


 


181
GCGGACAGCG TAAAGGGACG ATTCACCATT AGTAGAGACG ACGCAAAGAA TACTGTATAT


 


241
TTGCAGATGA ATTCGTTGAA GCCTGAGGAC ACCGCTGTCT ATTTTTGCGC GGCGCGACCG


 


301
AGGGGCGGTC TCTCCCCGAC ACCTCAAGCA TATGATTACT GGGGACAAGG GACCCAAGTC


 


361
ACTGTATCCA GTGCGGCCGC GAGCGGCAGC CTTGAACAAA AGCTGATAAG CGAGGAGGAT


 


421
CTCAATGGTG CTGCACATCA TCATCACCAT CACGGGGCAG CG,






and an exemplary sequence encoding EAT-1G4 could be:









(SEQ ID NO: 32)










1
GAAGTTCAGC TTGTAGAGTC CGGTGGGGGT CTTGTACAGC CCGGCGGGAG CTTGCGACTC



 


61
TCATGCGCTG CTTCCGGAAG CATTGCGACA ATAAATGATA TGGGTTGGTT TAGACAAGCC


 


121
CCCGGGAAGC AGCGTGACTG GGTCGCGACT ATTGTGAGTG ACGGCAGCAC GGCTTATGCG


 


181
GACTCAGTGA AAGGGAGATT TACGATTTCG CGAGATAACG CGAAAAACAC TGTATACCTG


 


241
CAGATGAATT CACTCAAGCC GGAAGATACA GCTGTGTATT ATTGTTCTGC CCGACGGCAC


 


301
TACGGACAGG GGACCCAGGT CACAGTCTCG AGCGCTGCCG CCAGTGGGTC ACTCGAGCAG


 


361
AAGCTGATAT CAGAGGAGGA CCTTAACGGT GCGGCGCACC ATCACCACCA TCATGGTGCG


 


421
GCG.






NetB is a heptameric beta-pore-forming toxin that forms single channels in planar phospholipid bilayers. The NetB activity is influenced by membrane fluidity and by cholesterol, which enhances the oligomerization of NetB and plays an important role in pore formation. NetB has high haemolytic activity towards avian red blood cells.
Neutralization of the NetB haemolytic activity by camelid VHH antibodies directed towards NetB is determined by measuring NetB-mediated lysis of chicken erythrocytes. The ability to neutralize the NetB haemolytic activity is assessed by pre-incubating a two-fold dilution series of the VHH antibodies (two wells per dilution, 5 μM starting concentration) with a constant amount of NetB toxin (20 μg recombinant NetB) for 30 minutes at 37° C. prior to the addition of 1% chicken erythrocytes. The non-toxic NetB variant W262A is included as a negative control as this variant displays no haemolytic activity. Positive control serum from rabbits immunized with the recombinant NetB (wild type NetB) is used, starting from a 1:4 dilution. After incubation at 37° C. for 1 hour, the plates are centrifuged to pellet intact red blood cells. The supernatants are transferred to a new 96 well plate and the A550 is determined. NetB activity is indicated by the increase in absorbance due to release of haemoglobin from the erythrocytes. The control serum is able to neutralize the haemolytic activity of NetB. VHH antibodies ENB-1F4 and ENB-1F10 have no effect on the NetB haemolysis. ENB-1B9 has intermediate inhibitory capacity, while ENB-1D11 and ENB-1A4 are able to neutralize the NetB haemolysis up to a 4- to 8-fold dilution (1.25 μM-0.625 μM VHHs).
Based on the above results, ENB-1A4 and ENB-1D11 are selected for further characterization. The peptide sequence of ENB-1A4 is:







(SEQ ID NO: 33)


EVQLVESGGGLVQAGGSLRLSCAASGSIFSTNVMGWYRQAPGKQREFVAG


 


ITIGGTARYPDSVKGRFTISRDNTQNTVYLQMNNLKPEDTAVYYCNAVLP


 


SDQRRWSWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA,






and the peptide sequence of ENB-1D11 is:








(SEQ ID NO: 34)


EVQLVESGGGLVQTGGSLRLSCTASGTIDMTYGLIWYRQAPGKERELVAS


 


IRRDGRTNYADSVKGRFTISIDNAKNSIHLQMNSLKPDDTARYYCNSPYH


 


ALWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHEIHHGAA.






An R56H mutant of ENB-1D11 (SEQ ID NO: 50) may also be generated to improve protease resistance of the antibody.
A person of skill in the art would recognize that, because of the redundancy of the genetic code, multiple nucleic acid sequences could encode the above peptides.
However, an exemplary sequence encoding ENB-1A4 could be:








(SEQ ID NO: 35)










  1
GAGGTACAAC TGGTTGAGAG TGGGGGTGGT TTGGTGCAAG CCGGAGGTTC CTTACGTTTG



 


 61
TCTTGCGCGG CTAGTGGGAG CATCTTTTCA ACAAACGTAA TGGGGTGGTA CCGCCAAGCC


 


121
CCAGGTAAGC AGCGGGAATT TGTGGCCGGG ATAACGATCG GAGGAACTGC GAGGTATCCT


 


181
GATAGTGTGA AAGGGCGTTT CACAATTAGT CGAGATAATA CACAGAATAC TGTCTATCTC


 


241
CAAATGAATA ATCTCAAGCC CGAAGACACA GCAGTTTATT ATTGTAATGC CGTTCTCCCC


 


301
TCTGATCAGC GTCGATGGAG CTGGGGACAA GGCACCCAGG TTACGGTTAG CAGCGCGGCA


 


361
GCGTCTGGTT CGCTCGAGCA AAAGCTCATA TCTGAGGAGG ACCTGAACGG GGCAGCCCAC


 


421
CATCACCACC ATCACGGAGC AGCT,






and an exemplary sequence encoding ENB-1G4 could be:









(SEQ ID NO: 36)










1
GAGGTACAGC TGGTGGAGTC CGGCGGTGGT TTGGTGCAAA CCGGGGGTAG TCTGCGGCTT



 


61
AGTTGCACGG CGTCTGGGAC AATAGACATG ACTTATGGTC TCATATGGTA CAGGCAAGCG


 


121
CCTGGGAAAG AGAGGGAACT CGTTGCGAGT ATCAGAAGGG ACGGCCGCAC AAATTACGCT


 


181
GATTCAGTGA AAGGGCGCTT CACTATCTCG ATCGATAATG CGAAAAACAG TATTCACCTT


 


241
CAAATGAACT CCCTTAAGCC CGATGATACC GCCAGGTATT ATTGCAACAG CCCATATCAC


 


301
GCACTTTGGG GTCAGGGTAC GCAGGTAACA GTGTCTAGTG CGGCAGCCTC TGGTAGTTTG


 


361
GAGCAAAAGT TGATAAGTGA GGAGGACTTA AATGGGGCGG CACATCACCA CCACCATCAT


 


421
GGGGCGGCT.






Example 8 Multifactorial Expression Cassettes
Expression cassettes may be designed and synthesized with different combination of the following components: promoter for expression, signal sequence for secretion, cell-wall anchor for secretion, at least one heterologous coding region encoding a desired biomolecule, and terminators for transcription termination. An expression cassette may contain multiple heterologous coding regions.
One representative expression cassette for the expression of camelid antibodies is:








(SEQ ID NO: 37)










  1
AAATTAAAAG GCTGGATTTT TTCGGCCTTT TTTTAGTGCA AATAATTATT TTTTACGTAT



 


 61
TTATATTATA GGGCTAATCA CTAAACTAAT AATTAGTGGT TGAAGCGCTG AAAATTTTCT


 


121
GCTATTTTAT TAATAGTTTG ATAATAAAAT AATGATATTT AATATAAAGA GGGATAAACG


 


181
AAATAATGAA ATCAACAACA AAGAAAATTC TTGCATCGTC GTTAGGGGTA GCTGGCGCAA


 


241
TGGCAATGGG CACGGTAACT GCAAAGGCTG ATACGACCGT TACGGTCAAT GCTGGCGATA


 


301
GTTTGAATGG GATTGCTCAA AAGTATAATG TTAGTGCGGA TGATATTGCA ACCGCTAATC


 


361
ACTTGCAAAA TAAAGAGTTG ATTTTTGTGG GACAAAAGTT GACAATTCCA ACCAAAGATA


 


421
AAAATGAAAC AGAAGTTCAA TTAGTTGAAA GTGGTGGTGG TTTAGTTCAA CCAGGTGGTA


 


481
GTTTACGTTT ATCATGTGCT GCAAGTGGTT CAATTGCAAC TATTAATGAT ATGGGTTGGT


 


541
TTCGTCAAGC ACCAGGAAAG CAACGTGATT GGGTTGCTAC TATTGTTAGT GATGGTTCAA


 


601
CTGCTTATGC TGATAGTGTT AAAGGTCGTT TTACTATTTC ACGTGATAAT GCTAAGAATA


 


661
CTGTTTACCT TCAAATGAAT AGTCTTAAGC CAGAAGATAC TGCAGTTTAC TATTGTTCAG


 


721
CTCGTCGTCA TTATGGTCAA GGTACTCAAG TTACTGTTAG TTCAGCTGCA GCTAGTGGTT


 


781
CATTAGAACA AAAATTAATT TCAGAAGAAG ATTTAAATGG TGCAGCTCAT CATCATCATC


 


841
ATCATGGTGC AGCTTACTTC CCTGGAAATT ACTATGTTGT GATGCATTAA TATTCCCTTT


 


901
CACCTCACCT TTAATAATTT AAATTAGTAA TTATCTTGCG CATCACAAAA GAGTGCTATA


 


961
TACTATTTCA GATTAGAAAG TTTTATGAGG GAGACAAATT G;






where nucleotides 1 to 185 represent a promoter sequence from the Cwls endopeptidase (SEQ ID NO: 41), nucleotides 186 to 262 encode the Cwls endopeptidase secretion signal (SEQ ID NO: 23), nucleotides 263 to 407 encode a LysM domain for attachment to extracellular polysaccharides, nucleotides 408 to 431 are a spacer sequence, nucleotides 432 to 854 encode VHH EAT-1G4 (SEQ ID NO: 30) and nucleotides 855 to 890 are another spacer sequence ending in a stop codon for translation termination.

A second representative expression cassette for the expression of camelid antibodies is:








(SEQ ID NO: 38)










   1
GAAGTACAAA GTTACTTTAA CTATAATGAA AAACAAGACA ATATAAAGAA AACAACATAT



 


  61
AAGGTTCAGT TCATAACTGA TTAGATTTAT AATAAATATT GTAAATCGGA CAAAAATAAA


 


 121
TTAATTTTCA ATTAATTCAA AAAAACCATA TTTTTTTCGT TTTGGCATAT TTGGATTTGC


 


 181
TACACTAAAG ATGATCAAGA AAGGGGAAAA GATAATCTTC AATCTTGTGT ACTTAGTTTG


 


 241
TTAATTAATT TATAAATTTA GGGAGGAAAC CTATCATGGA AAAGACTATG AAAAAGAAAG


 


 301
CTTTAGTTGC AACTACTGCT GTAGCCGGTA TTACTTTAGT AGGAGAGGTT ACTACTGTTC


 


 361
ATGCCGCTGA CAATGTACAA CAAGAAGTTC AATTAGTTGA AAGTGGTGGT GGTTTAGTTC


 


 421
AACCAGGTGG TAGTTTACGT TTATCATGTG CTGCAAGTGG TTCAATTGCA ACTATTAATG


 


 481
ATATGGGTTG GTTTCGTCAA GCACCAGGAA AGCAACGTGA TTGGGTTGCT ACTATTGTTA


 


 541
GTGATGGTTC AACTGCTTAT GCTGATAGTG TTAAAGGTCG TTTTACTATT TCACGTGATA


 


 601
ATGCTAAGAA TACTGTTTAC CTTCAAATGA ATAGTCTTAA GCCAGAAGAT ACTGCAGTTT


 


 661
ACTATTGTTC AGCTCGTCGT CATTATGGTC AAGGTACTCA AGTTACTGTT AGTTCAGCTG


 


 721
CAGCTAGTGG TTCATTAGAA CAAAAATTAA TTTCAGAAGA AGATTTAAAT GGTGCAGCTC


 


 781
ATCATCATCA TCATCATGGT GCAGCTAAGA AGATTACAGA AGATCAAAAA GCAACAGAAG


 


 841
GCGGTTATCA TATTGTTAAT GGATCTGTTG TAGATAAGCA GAATAACTTG GTTAGTGGTT


 


 901
GGACTGTTAA GAATGGTCAA ATGGTTGATC CTGAAGGTAA CGTTATCAAA ACAACAATGT


 


 961
CTACAGCCCA AGGTGTTACT ATTGAAAAAA ATAATAGCAA GTCCGGGAAT ACAAAGACAA


 


1021
ACATGATTCA AACTTCTTTA ACTATTGCTA ACAACAAGGC AACAACAAAC AAAGACAACC


 


1081
AGTTACCACA AACTGGCAAT TACAACAACA ATACAAAGGT ATTAGGATTA GCTGGTATTG


 


1141
CACTTGCATC TGCTTTAACT ATGTTTGGAT ACAAGAAGCG CCAACATAAC TAATTTTCTT


 


1201
ACTTGATGGG TTTCTAAATA AAAAATGGAC TACTTCAGCT CAAGGTAGCC CATTTTTATT


 


1261
ATTATAGTGA AGCAGTTATT CTTACGTATA ACCAGACTAA AATATAATAA AAATCTATTA


 


1321
TATATTAATC AACATCTCGG TTTAATCGTT AAAACTCCTC TGAGAGCTAA TTGTTAATAT


 


1381
TGAGTTGTAT AGT,






where nucleotides 1 to 275 represent a promoter sequence from chromosome segregation protein (SEQ ID NO: 42), nucleotides 276 to 383 encode a secretion signal from the chromosome segregation protein (SEQ ID NO: 22), nucleotides 384 to 806 encode VHH EAT-1G4 (SEQ ID NO: 30), nucleotides 807 to 1190 encode the cell wall anchor sequence from the chromosome segregation protein (SEQ ID NO: 22), and nucleotides 1191 to 1193 are a stop codon for translation termination.

A third representative expression cassette for the expression of camelid antibodies is:








(SEQ ID NO: 39)










   1
GCAATGCACA AGATGCTGAA ACAAAGGCAC AACAAAATGC AGATCAAGCT TCACCAGCTA



 


  61
ATATTCAAAA GGCACAAGAT GCTATTGCTA ATCAAGAAAC TCAAATTAGT AAAGACACCG


 


 121
ATGCTATTAA TGACGCTAAC AAAGCCGTTA GCGATGCACA AAGCACAGTT GATGCAGCGC


 


 181
AAAAAAAGTT AATGATGCAA CTACTGCTCG TGACAATCAA CAAAAGAATG TTGATACTGC


 


 241
TAGTGATGCA GTTAAGAATG CTCAAGCTAT TCTTGACAAC AGTGATCAGG CTAAAAAGGA


 


 301
AGCCCAAGAT GCTTTGAACA AGGCTAACCA AAAAGTTGCT GATGATACTA CTGCCGTTAA


 


 361
CAACAAACAA ACTGATGTTA ACAATGCAGC AGAAGCTAAG AAGAATGCAG ATGAGGCATT


 


 421
GAAGAACGCC AATGATGCGC AAACTTCTGC ACAAAAGAAT AAAGATGCTA AGCAAGCAAT


 


 481
TGCTGATGAG GCAAGTGTAG AAGTTCAATT AGTTGAAAGT GGTGGTGGTT TAGTTCAACC


 


 541
AGGTGGTAGT TTACGTTTAT CATGTGCTGC AAGTGGTTCA ATTGCAACTA TTAATGATAT


 


 601
GGGTTGGTTT CGTCAAGCAC CAGGAAAGCA ACGTGATTGG GTTGCTACTA TTGTTAGTGA


 


 661
TGGTTCAACT GCTTATGCTG ATAGTGTTAA AGGTCGTTTT ACTATTTCAC GTGATAATGC


 


 721
TAAGAATACT GTTTACCTTC AAATGAATAG TCTTAAGCCA GAAGATACTG CAGTTTACTA


 


 781
TTGTTCAGCT CGTCGTCATT ATGGTCAAGG TACTCAAGTT ACTGTTAGTT CAGCTGCAGC


 


 841
TAGTGGTTCA TTAGAACAAA AATTAATTTC AGAAGAAGAT TTAAATGGTG CAGCTCATCA


 


 901
TCATCATCAT CATGGTGCAG CTGCTGAAAA GGAAGCTAAA GATAATGGCT ACCATATCGA


 


 961
AAATAACCAA GTTGTTGACG CTAAAGGTAA TAGTGTCAAT GGCTGGACAG TTAAGGGCAA


 


1021
CCAAATTGTT AGTCCAACTA ATGCTACTGT AGATCCCGCT GTTTCTGTAA CCACCAATGT


 


1081
CAATGTTGAT AGTAAAGGTC AAGTACAACC ACAAACTAGT GTTACTGCTA ATAGTGTTAA


 


1141
GACTGTAGCT GCAACTGAAT CAGCAAATCC AGTAGCAACT ACTACTGTGC AAACCCGCGA


 


1201
ACAATACAAG CAACAATTGA AGAGCAATAA TCAATTACCA CAAACTGGTA ATAATGATAG


 


1261
TGCTGTTCTT TCACTTGCTG GAGTAGCACT TGCAGCAATG TTGAGTTTGT TCGGTATTAA


 


1321
GAAACGTGAA TACTAATTTA GAAAATGTAA GTATTATTAT GTAAAAAGGT TCAACCAAAT


 


1381
TGGCTGAACC TTTTTGTCTA AAATTTAAGG AGAAGTTTT,






where nucleotides 1 to 313 represent a promoter sequence from Chromosome Partition Protein Smc (SEQ ID NO: 43), nucleotides 314 to 499 encode a secretion signal from Chromosome Partition Protein Smc (SEQ ID NO: 40), nucleotides 500 to 922 encode VHH EAT-1G4 (SEQ ID NO: 30), nucleotides 923 to 1333 encode the cell wall anchor sequence from Chromosome Partition Protein Smc (SEQ ID NO: 40), and nucleotides 1334 to 1336 are a stop codon for translation termination.

Many other combinations of the disclosed expression cassette elements are also possible.
Example 9: Anti-infective Molecules Derived from Bacillus 
Certain Bacillus strains may also be used as DFM, for example for poultry. Over one hundred Bacillus isolates are collected from chicken cecum. The chickens are sourced with the state of Indiana, USA. Isolated strains are selected to withstand 100° C. for 10 minutes for development as pellet-stable products. The isolated strains are confirmed to be Bacillus using 16S rRNA sequencing. The isolates are tested for antimicrobial activity against various poultry pathogens such as Salmonella Enteritidis, avian pathogenic E. coli, Clostridium perfringens, Enterococcus cecorum and Campylobacter jejuni. Different isolates display different antimicrobial activities against these pathogens. Of all the isolates, five isolates are cytotoxic for all the pathogens tested. All 5 isolates are isolated from samples collected from the same study, suggesting that they may potentially be clonal.—Two isolates, D24 and D72, are selected for further characterization.
The two selected isolates (D24 and D72) are sequenced by ILLUMINA® sequencing and determined to most likely be Bacillus velezensis. The genomic sequences are analyzed for potential bacteriocins, antimicrobial peptides and digestive enzymes. The isolates contain several full loci potentially encoding for antimicrobial peptides and bacteriocins. Considering the feasibility of engineering these antimicrobial peptides and bacteriocins into an L. reuteri expression system, five ribosomal bacteriocins are selected for further study. Based on the sequence homology, the five Bacillus bacteriocins are identified to be
Antimicrobial Peptide LCI:


    
    
        AIKPVPSPNGIFAASFELNGTTWIFKYKYYDSSKGYWVGIYESVDK (SEQ ID NO: 44); Circularin A:
    
    


LASTLGISTAAAKKAIDIIDAASTIASIISLIGIVTGAGAISYAIVATAKTMIKKYGKK YAAAW (SEQ ID NO: 45); Lanthipeptide 3 (Plantaricin C): EFSGGGGAEQRGISQGNDGKLCTLTWECGLCPTHTCWC (SEQ ID NO: 46); Lanthipeptide 5 (Lichenicidin A2): SDATPMTVTPTTITIPISLAGCPTTKCASIVSPCN (SEQ ID NO: 47); and Lanthipeptide 6: SEATPMTVTPTTITIPISLAGCPTTKCASIVSPCND (SEQ ID NO: 47). These bacteriocins have been previously shown to kill various Gram-negative and Gram-positive pathogens.
These Bacillus-derived anti-infective peptides may be incorporated into an expression cassette and expressed recombinantly. Recombinant Bacillus-derived anti-infective peptides may be expressed alone or (without limitation) one or more of the other anti-infective peptides or phage disclosed in Example 2, the lytic enzymes disclosed in Example 6, or antibodies such as those disclosed in Example 7.
Example 10: In Vivo Effects of L. reuteri 
An identified strain can effectively function as a direct feed microbial only if the strain is able colonize the host gastrointestinal tract. The ability of L. reuteri strains to colonize chickens is assessed following different routes of administration.
L. reuteri is administered orally to 10-day old chicks. At various times post inoculation, chicks are sacrificed and the presence of L. reuteri is measured in the crop, small intestine, and cecum. Strain 3632 is able to colonize all three tissues and remain detectable for at least 18 days.
L. reuteri is administered by in ovo inoculation (i.e. injection into the egg) 3 days before hatching. Strain 3632 is found in the tissues of chicks at 3 and 7 days post hatching, while colonization of a control strain declines after day 3.
L. reuteri is administered in ovo by spraying a liquid containing bacteria onto the egg surface. Again, Strains 3630 and 3632 are able to colonize and remain present in chicks 3 and 7 days post hatching. Inoculation by spray is just as efficient as in ovo injection but requires less manipulation of the incubating egg.
Recombinants strains 3630 and 3632 carrying expression cassettes comprising one of the antibodies of Example 7 or the PlyCM lytic enzyme are administered by in ovo inoculation. The recombinant strains are able to efficiently colonize chicken gastrointestinal tracts at comparable levels to unmodified parental strains. Thus, genetic manipulation as contemplated herein does not alter the ability of the modified strains to act as DFM.
Chicks carrying L. reuteri strains or modified strains are assessed for their susceptibility to necrotic enteritis. Chicks are inoculated with L. reuteri live expression systems wherein the expression cassettes contain either a single chain antibody or a lytic enzyme. Inoculation is done orally by providing chicks drinking water containing 108 recombinant L. reuteri. 
An experiment is performed as given in Table 2, with 40 one-day-old chicks placed in each of ten groups. Group 1 chicks served as the untreated and unchallenged controls. Group 2 chicks served as untreated challenge controls. Group 3 represents challenged chicks given standard antibiotic therapy. Groups 2-10 each received 2×108 CFU of C. perfringens on each of days 17 and 18. Groups 4-9 each received prophylactic inoculation with L. reuteri continuously from day 1. Group 10 received only therapeutic doses of L. reuteri on days 18-21. On day 21 chicks are sacrificed and small intestinal tissues are examined for the presence of lesions. Survival of chicks within each treatment group is also recorded.
As shown in Table 2, the four antibodies can each ameliorate disease to some degree. A combination of strains may be more effective than any single strain.







TABLE 2







Antibodies produced by recombinant L. reuteri can treat necrotic enteritis.


















Lesion
reduction
Mortality
Reduction in


Group
Strain
Ab
Challenge
score (avg)
in lesions
(avg)
mortality

















1
none
none
none
0
n/a
0
n/a


2
none
none
Yes
1.80
n/a
27.5
n/a


3
none
none
Yesa
0.375
79.17%
2.5
90.91%


4
3630
1G4
Yes
1.35
25.00%
22.5
18.18%




1F2









1D11









1A4







5
3630
1G4
Yes
0.5
72.22%
10
63.64%


6
3630
1F2
Yes
0.95
47.22%
20
27.27%


7
3630
1D11
Yes
1.25
30.56%
25
 9.09%


8
3630
1A4
Yes
1.45
19.44%
22.5
18.18%


9
3630
1D11
Yes
0.675
62.50%
0
100.0%



3632
1D11







10
3630
1G4
Yes
1.275
29.17%
17.5
36.36%




1F2









1D11









1A4





aTreated with BMD. Bacitracin methylene disalicylate