CN117730146A - Production of lytic phages - Google Patents

Abstract

The present invention relates to a producer bacterial cell for producing lytic phage particles or a lytic phage-derived delivery vehicle, said producer bacterial cell stably comprising at least one phage structural gene and at least one phage DNA packaging gene, said phage structural gene and phage DNA packaging gene being derived from a lytic bacterial phage, wherein the expression of at least one of said phage structural genes and/or at least one of said phage DNA packaging genes in said producer bacterial cell is controlled by an induction mechanism.

Description

Production of lytic phages

Technical Field

The present invention relates to bacterial cells for producing lytic phage particles and methods of using such bacterial cells.

Background

Lytic bacteriophages (phages) are a self-replicating virus that is able to infect and lyse its specific host bacteria. Lytic phages are considered as an alternative solution against antimicrobial resistant pathogens due to their host specificity and non-toxicity.

However, lytic phages are naturally not stably maintained in the genome and/or as episomes/in strains, which makes their production on an industrial scale extremely complex.

Furthermore, most current phage or phage-derived delivery vehicle generation methods mean the use of bacterial species or strains as natural hosts for the phage as production cells. Such methods may be dangerous as a result when such bacterial cells are pathogenic, for example when they produce toxins. Furthermore, many bacterial species cannot be easily manipulated, for example, because of their growth conditions, or because these bacteria have no effective genetic tools.

Thus, there is a need for methods that allow for safe, easier and efficient production of lytic phages or lytic phage-derived particles.

The inventors believe that the phage may be regarded as a more or less large genetic circuit, the final output of which is to produce more phage particles. To this end, whether the phage is lytic, temperate or chronic (e.g., filamentous phage such as M13), the inventors believe that the information encoded in its genome can be broadly classified as:

genes specific for insertion/excision (for temperate phages).

Genes specifically for DNA replication, RNA transcription, etc. In fact, for example, some lytic phages encode their own RNA or DNA polymerase. Some genes modify the RNA polymerase of the host to be able to work beyond the terminator and if the prophage sequence is present in plasmid or linear plasmid form, some other genes are involved in the isolation of the prophage sequence.

Genes specifically used to package the newly synthesized phage genome into the newly synthesized phage capsid: termination enzymes and accessory proteins, ligases, and the like.

-structural genes specific for the construction of protein capsids for DNA: in addition to stringent structural genes such as capsid genes, tape measures, fibers, substrates, etc., many other genes are needed to assemble the components (chaperones, proteases) and proteins that can be packaged within the capsid, either as scaffolds or as leader proteins injected into the cell (e.g., RNA polymerase of phage N4 or some small leader proteins in other phages (minor pilot protein)).

Genes associated with defenses against host anti-phage mechanisms, degradation/modification of host elements to complete the cleavage cycle, super-rejection mechanisms or genes beneficial to the host.

DNA packaging and structural gene classes are deeply linked in that the packaging mechanism recognizes the pre-assembled heads and the DNA to be packaged in these heads, starting and stopping DNA packaging.

The inventors hypothesize that by extracting and differentiating all the modules defined above, it is possible to construct a system containing all the excision/insertion, replication and regulatory elements from non-lytic phages and encoding packaging/structural elements of lytic phages, since they can be considered as independent genetic modules as the inventors believe.

Treating them as independent genetic modules may also allow the construction of a system containing only the desired lytic phage structures and/or regulatory elements to be produced under the control of a primary regulatory element (e.g., an inducible repressor) that may not originate from a phage. For example, only the structural operon and DNA packaging machinery of lytic phage can be placed under the control of a repressor that responds to small molecules or physical/chemical signals (LacI, araC, phlF, λci, etc.), triggering the production of all elements required to produce a pure mature lytic phage delivery particle (phage or packaged phagemid). This "reduced" version of the phage genome is stably maintained in the bacterial production strain.

Summary of The Invention

The present invention stems from the unexpected discovery by the inventors that by exchanging the structural operon of an Escherichia coli (Escherichia coli) producing strain encoding a system that produces pure Lambda-packaged phagemid with structural elements of a strictly lytic phage, such as a T7 phage, the assembly and packaging of pure heterologous lytic phagemid particles can be driven upon supplementation with a plasmid containing the correct packaging signal (LTR for T7 phage). Thus, the inventors herein demonstrate that packaged phagemids can be structurally based on T7 lytic phage production, but are regulated and maintained in a lysogenic state by the Lambda prophage mechanism in E.coli producing strains.

The inventors have also shown that the structural operon of Propionibacterium freudenreichii (P.freudenreichii) prophage can be exchanged with the structural operon of the lytic phage of the skin bacillus acnes (C.acnes) strain. In this way, the inventors have shown that the assembly and packaging of pure Clostridium acnes phagemid can be driven by exchanging the structural operon of the Prophage of Propionibacterium freudenreichii with that of the lytic phage of the dermatophyte acnes strain.

The present invention thus relates to a producer bacterial cell for producing lytic phage particles or a lytic phage-derived delivery vehicle, said producer bacterial cell stably comprising at least one phage structural gene and at least one phage DNA packaging gene, said phage structural gene and phage DNA packaging gene being derived from a lytic bacteriophage,

wherein the expression of at least one of said phage structural genes and/or at least one of said phage DNA packaging genes in said producer bacterial cell is controlled by an induction mechanism.

The invention also relates to a method for producing lytic phage particles or a lytic phage-derived delivery vehicle comprising:

(a) Providing a producer bacterial cell of the invention, and

(b) In the production bacterial cell, inducing expression of the at least one of the phage structural genes and the at least one of the phage DNA packaging genes, and assembly of products expressed by the at least one phage structural gene and the at least one phage DNA packaging gene, thereby producing lytic phage particles or a lytic phage-derived delivery vehicle.

Another object of the present invention relates to a hybrid helper phage system comprising:

(i) At least one phage DNA packaging gene derived from a lytic bacteriophage,

(i') at least one phage structural gene derived from said lytic bacteriophage,

(i ") optionally, at least one phage gene involved in phage regulation derived from said lytic bacteriophage, and

(ii) At least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation,

wherein the genes (i), (i') and (ii) are contained in separate nucleic acid molecules or separate nucleic acid molecules, and

wherein the hybrid helper phage system does not comprise any expressed phage structural genes derived from the non-lytic bacteriophage.

Detailed Description

Production of bacterial cells

The present invention relates to a producer bacterial cell for producing lytic phage particles or a lytic phage-derived delivery vehicle, said producer bacterial cell stably comprising at least one phage structural gene derived from a lytic bacteriophage and at least one phage DNA packaging gene,

wherein the expression of at least one of said phage structural genes and at least one of said phage DNA packaging genes in said producer bacterial cell is controlled by an induction mechanism.

As used herein, the term "phage particle" refers to a functional or nonfunctional (e.g., non-proliferative and/or replicative) viral particle.

As used herein, the term "lytic phage particle" refers to a particle derived from a phage that is not naturally stable in the genome and/or remains as an episome/in a strain and thus has a strictly lytic (as opposed to lysogenic) lifestyle, i.e. the infection process always ends with lysis of the strain of interest.

As used herein, the term "lytic phage-derived delivery vehicle" refers to any means that allows the payload to be transferred into bacteria and derived from lytic bacteriophage. In the context of the present invention, the term "lytic phage-derived delivery vehicle" further encompasses lytic phage-derived particles that do not comprise any payload but are capable of targeting bacterial cells.

Lytic phage-derived delivery vehicles may refer to lytic phage-derived scaffolds and may be obtained from natural, evolved or engineered lytic phage.

Bacterial cells

The producer bacterial cells of the invention may belong to any bacterial species or strain, in particular as defined under the "target bacteria" section below.

However, the producer bacterial cell is preferably a non-pathogen bacterial cell. Still preferably, the producer bacterial cell is an easily manipulable bacterial cell.

By "readily manipulable" is meant herein that bacterial cells may be cultured and/or modified using well known techniques.

In certain preferred embodiments, the producer bacterial cell is an E.coli bacterial cell. Alternatively, the producer bacterial cell may be a Bacteroides (bacterioides) bacterial cell, more particularly a Bacteroides thetaiotaomicron (Bacteroides thetaiotaomicron) bacterial cell, a propionibacterium freudenreichii (Fusobacterium) bacterial cell or a Streptococcus (Streptococcus) bacterial cell. In a particular embodiment, the producer bacterial cell is a propionibacterium freudenreichii bacterial cell.

The production bacterial cells of the present invention may be obtained by any technique well known to the skilled person, in particular by introducing said phage structural genes and phage DNA packaging genes derived from lytic bacterial phages into the bacterial cells by any technique well known in the art.

The producer bacterial cells of the invention can generally be obtained by homologous recombination or recombinant engineering involving, for example, MAGE (Wannier et al Recombineering and MAGE. Nat Rev Methods Primers, 7 (2021)), e.g., using CRISPR, TALEN, meganuclease and/or zinc finger techniques, or site-specific recombination with phage integrase, PASTE (Ioanidi et al, drag-and-drop genome insertion without DNA cleavage with CRISPR-directed integers. Biorxiv 2021.11.01.466786 (2021) doi 10.1101/2021.11.01.466786), or transposon-related CRISPR-Cas system (Ma et al, trends Microbiol 29,565-568 (2021)).

Phage DNA packaging gene and phage structural gene

The production bacterial cells of the present invention stably comprise at least one phage structural gene derived from a lytic bacteriophage and at least one phage DNA packaging gene.

By "stably included" or "stably included (stably comprising)" is meant herein that the production bacterial cell retains the phage structural gene and phage DNA packaging gene, either incorporated into its chromosome or on an episome that is typically maintained in the cell by selection (e.g., with an auxotroph, or drug resistance marker). Each gene stably contained by the producer bacterial cell may be independently located on a plasmid, on a helper phage, or integrated into the producer bacterial cell chromosome.

In certain embodiments, the producer bacterial cell stably comprises at least two, 3, 4, or all phage structural genes derived from the lytic bacteriophage, and at least one phage DNA packaging gene derived from the lytic bacteriophage.

In certain embodiments, the producer bacterial cell stably comprises at least one phage structural gene derived from the lytic bacteriophage and at least two or all phage DNA packaging genes derived from the lytic bacteriophage.

In particular embodiments, the producer bacterial cell stably comprises at least two, 3, 4 or all phage structural genes derived from the lytic bacteriophage and at least two or all phage DNA packaging genes derived from the lytic bacteriophage.

In particular embodiments, the producer bacterial cell stably comprises all phage structural genes derived from the lytic bacteriophage and all phage DNA packaging genes derived from the lytic bacteriophage.

By "phage structural gene" is meant herein a gene from a bacteriophage that is involved in the construction of the capsid of a bacteriophage protein. Phage structural genes include genes encoding phage structural elements; genes encoding phage proteins involved in the assembly of phage structural elements; and genes encoding phage proteins packaged within the capsid as scaffolds or as lead proteins to be injected into targeted bacterial cells.

Phage building elements are well known to the skilled person and depend on the type of bacteriophage from which they are derived. Phage building elements can be proteins, but also RNAs (e.g., phi29 of some phages, such as bacillus subtilis (Bacillus subtilis), encodes a structural scaffold made of RNA). Phage structural elements generally include capsid proteins, tape proteins, fibers, substrate proteins, tail sheath proteins, whisker proteins, ornamental proteins, and the like.

Phage proteins involved in the assembly of structural elements are well known to the skilled person and depend on the type of bacteriophage from which they are derived and optionally on the structural elements encoded by other phage structural genes. Phage proteins involved in the assembly of structural elements generally include phage chaperones and phage proteases.

Phage proteins that are packaged within the capsid as scaffolds or as lead proteins to be injected into target host cells are well known to the skilled artisan and depend on the type of bacteriophage from which they are derived. Examples of such phage proteins are RNA polymerase or small leader proteins from phage N4.

As the skilled artisan will appreciate, the presence of a particular phage structural gene in a production bacterial cell of the invention will depend on the bacteriophage from which the phage structural gene is derived.

By "phage DNA packaging gene" is meant herein a gene from a bacteriophage that participates in packaging the bacteriophage genome into the bacteriophage capsid. Phage DNA packaging genes are well known to the skilled artisan and include genes encoding phage terminases, genes encoding phage accessory proteins, genes encoding phage ligases, genes encoding phage exonucleases involved in DNA packaging, and genes encoding phage endonucleases involved in DNA packaging.

In certain embodiments, the producer bacterial cell further stably comprises at least one gene derived from the lytic bacteriophage involved in phage regulation.

By "genes involved in phage regulation" is meant herein phage genes involved in phage interaction with a host. Examples of genes involved in phage regulation include phage genes encoding the primary repressor, phage genes encoding anti-stop proteins, phage genes involved in transcription (such as phage genes encoding RNA polymerase), phage genes involved in superrejection mechanisms, phage genes involved in defense against host anti-phage mechanisms, phage genes involved in degradation and/or modification of host elements, e.g., to complete the lytic cycle, and phage genes that are beneficial to the host.

In particular embodiments, the producer bacterial cell stably comprises phage genes derived from the lytic bacteriophage that are involved in defense against host anti-phage mechanisms.

In another specific embodiment, the producer bacterial cell stably comprises a bacteriophage gene involved in transcription, such as a bacteriophage gene encoding an RNA polymerase, derived from the lytic bacteriophage.

In a specific embodiment, said phage structural gene and phage DNA packaging gene derived from said lytic bacteriophage, and optionally said gene derived from said lytic bacteriophage involved in phage regulation, are contained in at least one plasmid, chromosome and/or helper phage. In a specific embodiment, the phage structural gene and phage DNA packaging gene derived from the lytic bacteriophage, and optionally the gene derived from the lytic bacteriophage involved in phage regulation, are comprised in at least two separate nucleic acid molecules, in particular at least two plasmids, chromosomes, helper phage, or a combination thereof.

In a specific embodiment, the phage structural genes and phage DNA packaging genes derived from the lytic bacteriophage, and optionally the participating phage controls, genes derived from the lytic bacteriophage are included in a hybrid helper phage system as defined below.

In a specific embodiment, the phage structural gene and phage DNA packaging gene derived from the lytic bacteriophage and optionally the gene derived from the lytic bacteriophage involved in phage control are contained in a helper phage.

Induction mechanism

In the context of the present invention, the expression of at least one of said phage structural genes and/or at least one of said phage DNA packaging genes in said producer bacterial cells as defined in the "phage DNA packaging genes and phage structural genes" section above is controlled by at least one induction mechanism.

In a specific embodiment, the expression of at least one of said phage structural genes, in particular at least two, at least three or all of said phage structural genes in said producer bacterial cells is controlled by at least one induction mechanism, in particular by one induction mechanism.

In a specific embodiment, the expression of at least one, in particular at least two, at least three or all of said phage DNA packaging genes in said producing bacterial cells is controlled by at least one induction mechanism, in particular by one induction mechanism.

In certain embodiments, the same induction mechanism controls the expression of at least one of the phage structural genes and at least one of the phage DNA packaging genes.

In alternative embodiments, the expression of at least one of the phage structural genes and the expression of at least one of the phage DNA packaging genes are controlled by different induction mechanisms.

By "induction mechanism" is meant herein a mechanism encoded by a gene or set of genes contained, in particular stably contained, in the producer bacterial cell, which is capable of inducing their controlled gene expression in response to a given trigger.

In certain embodiments, the induction mechanism further controls the copy number of the at least one of the phage structural genes and/or the at least one of the phage DNA packaging genes. In other words, in a specific embodiment, the induction mechanism further controls the replication of said at least one of said phage structural genes and/or said at least one of said phage DNA packaging genes, in particular the replication of a nucleic acid molecule carrying said at least one of said phage structural genes and/or said at least one of said phage DNA packaging proteins.

In certain embodiments, the induction mechanism further controls assembly of products expressed by the at least one of the phage structural genes and the at least one of the phage DNA packaging genes.

Examples of such induction mechanisms include:

an induction system based on protein repressors or activators, which react to small molecules (e.g. sugars, quorum sensing molecules, gases, synthetic molecules, peptides, amino acids, metabolites, etc.), physical signals (temperature, pressure, etc.), chemical signals (osmotic pressure, pH, etc.), biological signals (cell density, DNA damage, etc.); these systems may be activated by secondary proteins such as orthogonal RNA polymerase or sigma factors.

Activating or repressing the protein degradation system due to transcription of the promoter.

RNA-based induction systems such as aptamers that react to the above signals, such as RNAi, CRISPRi, fulcrum systems, riboswitches, etc.

-one or more nucleic acids comprising at least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation.

In particular embodiments, the induction mechanism comprises at least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication, and/or phage regulation.

Thus, in particular embodiments, the producer bacterial cell further comprises at least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication, and/or phage regulation.

Genes involved in phage excision/insertion, phage DNA replication and/or phage regulation

By "genes involved in phage excision/insertion" is meant herein genes from lysophage involved in excision of the phage (present as a prophage) from the genome or episome of the bacterial cell and/or insertion of the phage (present as a prophage) into the genome or episome of the bacterial cell.

By "genes involved in phage DNA replication" is meant herein genes from lysophage involved in phage DNA replication mechanisms. Examples of genes involved in phage DNA replication include genes encoding DNA polymerase and genes involved in isolation thereof if the prophage sequence is present in plasmid or linear plasmid form.

By "genes involved in phage regulation" is meant herein phage genes involved in phage interaction with a host. Examples of genes involved in phage regulation include phage genes encoding the primary repressor, phage genes encoding anti-stop proteins, phage genes involved in the superrepulsion mechanism, phage genes involved in the defense host against phage mechanisms, phage genes involved in degradation and/or modification of host elements, e.g., to complete the lytic cycle, and phage genes that are beneficial to the host.

In the context of the present invention, the genes involved in phage excision/insertion, phage DNA replication and/or phage regulation are not DNA packaging genes or structural genes as defined above.

In a preferred embodiment, the production bacterial cells of the present invention comprise at least one gene, preferably all genes, derived from a second type of bacteriophage involved in phage excision/insertion; at least one gene, preferably all genes, derived from a non-lytic bacteriophage involved in phage DNA replication; and/or at least one gene, preferably all genes, derived from a non-lytic bacteriophage involved in phage regulation.

In the context of the present invention, the producer bacterial cells do not comprise genes derived from lytic bacterial phages involved in phage excision/insertion and/or phage DNA replication.

In certain embodiments, the genes derived from the non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication, and/or phage regulation are contained in at least one plasmid, chromosome, and/or helper phage. In a specific embodiment, the genes derived from the non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation are comprised in at least two separate nucleic acid molecules, in particular at least two plasmids, chromosomes, helper phage or combinations thereof.

In a specific embodiment, the genes derived from the non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation are comprised in a hybrid helper phage system as defined below.

In a specific embodiment, the genes derived from the non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation are comprised in a helper phage system, more particularly on the same helper phage system as the phage structural genes and phage DNA packaging genes derived from the lytic bacteriophage and optionally the genes involved in phage regulation derived from the lytic bacteriophage.

In particular embodiments, the producer bacterial cell is from the same bacterial species or strain from which the non-lytic bacteriophage is derived and/or to which the non-lytic bacteriophage is targeted.

In a more specific embodiment, the producer bacterial cell is an E.coli bacterial cell. In another specific embodiment, the producer bacterial cell is a propionibacterium freudenreichii bacterial cell.

Other elements

In a particular embodiment, the producer bacterial cell of the invention further comprises at least one gene involved in the transcription of phage RNA.

By "gene involved in phage RNA transcription" is meant a gene from a temperate or lytic phage involved in phage RNA transcription machinery. Examples of such genes include genes encoding phage RNA polymerase and phage genes encoding proteins that modify the RNA polymerase of the host that are generally capable of exceeding terminator operation.

Bacteriophage and genes derived from bacteriophage

By "gene derived from a bacteriophage" is meant herein that the sequence of the gene is obtained from the bacteriophage, which is optionally modified, recoded and/or optimized compared to the sequence originally present in the bacteriophage. For example, the sequences may be recoded for codon exchange or optimization or prevention of recombination.

Bacteriophages are obligate intracellular parasites that multiply within bacteria by the selection of some or all of the host biosynthetic machinery (co-opt). Alternatively, some bacteriophages (lytic bacteriophages) are not naturally stable in the bacterial genome and/or as episomes in bacteria and cause bacterial lysis. Most phages range in size from 24-200nm in diameter. Phages contain nucleic acids (i.e., genomes) and proteins, and can be encapsulated by lipid membranes. Depending on the phage, the nucleic acid genome may be DNA or RNA, and may exist in circular or linear form. The size of the phage genome varies depending on the phage. The simplest phage has a genome that is only a few thousand nucleotides in size, whereas more complex phages can contain more than 100,000 nucleotides in their genome, and in rare cases more than 1,000,000. The number and amount of individual types of proteins in a phage particle will vary depending on the phage.

By "lytic phage" is meant herein a bacteriophage that infects bacteria or archaea, and which is naturally unable to be stably maintained in the genome and/or as an episome/in a strain, and thus has a strictly lytic (as opposed to lysogenic) lifestyle, and causes lysis and destruction of bacteria or microbial cells after phage replication. Once the cells are destroyed, phage progeny can seek new host cells (e.g., bacteria) to infect. The skilled artisan knows which of the bacteriophages listed below are lytic phages.

In particular embodiments, the non-lytic bacteriophage is a temperate bacteriophage, a filamentous bacteriophage, or a pseudolysogenic bacteriophage.

By "temperate bacteriophage" or "lysogenic bacteriophage" is meant herein a bacteriophage that infects bacteria or archaea, which can be stably maintained in the genome and/or as an episome/in a strain, and which in its lysogenic state accompanies cell replication without production of viral particles. It is well known to the skilled person which bacteriophages are temperate phages among the bacteriophages listed below.

"filamentous phage" is herein intended to mean a bacteriophage characterized by having a single-stranded DNA genome surrounded by a long protein capsid cylinder. Generally, bacteria infected with filamentous phage do not lyse during the life cycle and replication of the phage, but rather experience a decrease in growth rate as phage particles are secreted from the membrane. It is well known to the skilled person which bacteriophages are filamentous phages among the bacteriophages listed below.

By "pseudolysogenic phage" is meant herein a bacteriophage in a host cell in a stage of arrested development that neither has multiplication of the phage genome (as in lytic development) nor its replication in synchronization with the cell cycle and stable maintenance in the cell line (as in lysogenic), which takes place without degradation of the viral genome, allowing subsequent restart of viral development.

In a particular embodiment, the lytic bacteriophage is selected from the group consisting of lytic bacteriophages of the order of the end phage (Order Caudovirales) and/or non-lytic bacteriophages are selected from the group consisting of non-lytic bacteriophages of the order of the end phage based on the classification by Krupovic et al (Krupovic et al, arch virol.2016Jan;161 (1): 233-47):

Myophagidae (Myoviridae) (such as, without limitation, the following genera: cp220 virus, cp8 virus, ea214 virus, felixo1 virus, moogle virus, susp virus, hp1 virus, P2 virus, kay virus, P100 virus, silvia virus, spo1 virus, tcarboma virus, twart virus, cc31 virus, jd18 virus, js98 virus, kp15 virus, moon virus, rb49 virus, rb69 virus, S16 virus, schizot4 virus, sp18 virus, T4 virus, cr3 virus, se1 virus, V5 virus, abouo virus, agate virus, agrican357 virus, ap22 virus, arv virus, B4 virus, bastille virus Bc431 virus, bep 78 virus, bsepmu virus, biquarta virus, bxz virus, cd119 virus, cp51 virus, cvm virus, eah virus, el virus, hapuna virus, jimmer virus, kpp10 virus, M12 virus, machina virus, martha virus, msw3 virus, mu virus, myohalo virus, nit1 virus, P1 virus, pakpuna virus, pbena virus, phikz virus, rheph4 virus, rsl2 virus, rslena virus, securda 5 virus, sep1 virus, spn3 virus, svuna virus, tg1 virus, vhml virus and Wph virus

Podovidae (Podovidae) (such as, without limitation, fri1 virus, kp32 virus, kp34 virus, phikmv virus, prado virus, sp6 virus, T7 virus, cp1 virus, P68 virus, phi29 virus, nona33 virus, pocj virus, tl2011 virus, bpep 22 virus, bpp virus, cba41 virus, dfl virus, ea92 virus, epsilon15 virus, F116 virus, G7c virus, jwalpha virus, kf1 virus, kpp25 virus, lit1 virus, luz24 virus, luz7 virus, N4 virus, nonana virus, P22 virus, page virus, phieco32 virus, prtb virus, sp58 virus, una virus and Vp5 virus)

Longfoot phage (Siphoviridae) (such as, without limitation: cam virus, lika virus, R4 virus, acacian virus, cooper virus, pg1 virus, pipiffish virus, rosebus virus, brujita virus, che9C virus, hawkeye virus, lot virus, jersey virus, K1g virus, sp31 virus, lmd virus, una virus, bongo virus, rey virus, butters virus, charlie virus, redi virus, baxter virus, nyphadora virus, bignuz virus, fishe burn virus, phayonce virus, kp36 virus, rogue1 virus, rtp virus, T1 virus Tls virus, ab18 virus, amigo virus, anatole virus, andromeda virus, attis virus, barnyard virus, bernal13 virus, biseptima virus, bron virus, C2 virus, C5 virus, cba181 virus, cbast virus, ceci virus, che8 virus, chi virus, cjw virus, corndog virus, crones virus, D3112 virus, D3 virus, decurro virus, demoshenes virus, doucette virus, E125 virus, eiau virus, ff47 virus, gaia virus, giles virus, tls virus, ab18 virus, amigo virus, anatole virus, andromeda virus, attis virus, barnyard virus, bernal13 virus, biseptima virus, bron virus, C2 virus, C5 virus, cba181 virus, cbast virus, ceci virus, che8 virus, chi virus, cjw virus, corndog virus, crones virus, D3112 virus, D3 virus, decurro virus, demosghenes virus, doucette virus, E125 virus, eiau virus, ff47 virus, gaia virus, giles virus, tria virus, trigintaduo virus, vegas virus, vendetta virus, wbeta virus, wildcat virus, wizard virus, woes virus, xp10 virus, ydn virus and Yua virus

Ekkaman virus family (Ackermannviridae) (such as, without limitation, the following genera: ag3 virus, limestone virus, cba120 virus and Vi1 virus)

In particular embodiments, lytic and/or non-lytic bacteriophages are not part of the end phage purpose, but are derived from a source having an unspecified purpose such as, without limitation, the multi-layered phage family (tectviridae) (such as the following genera: the genus A, B, cover phage (Cortovirus), the family of filoviridae (Inoviridae), the family of filoviridae (such as the genus Protovora (fibre virus), the family of zonal phage (Habenivirus), the family of filoviruses (Inovirus), the genus Linear phage (Lineavirus), the genus short-rod phage (Plactrovirus), the genus MarshAN_SNe (Saetevirus), the genus bat phage (Vesperovirus), the family of vesicular (Cytosviidae) (such as the genus Cytosviidae (Cytosviidae)), the family of smooth phage (Levidae) (such as the genus Isovides (Allevirus)), the family of smooth phage (Levidae), the genus Picoviridae (Levidae) (such as the genus Alvovirus), the genus Phamygdavirus (Phamivirus) and the genus Phamygdalidae (Phillips), the genus Phamygdaliotavirus (Phillips), the genus of Picoviridae (Phillips), the genus (Phillips) and the genus of the genus Phalamitis (Phillips).

In particular embodiments, the archaebacteria targeted by lytic and/or non-lytic bacteriophages is not part of a tailed phage of interest, but is derived from a family of unspecified interest such as, without limitation, the family of bottle-forming viruses (Ampulloviridae), the family of mini spindle-forming phages (Fuseloviridae), the family of globoviridae (globoviridae), the family of microdroplets (Guttaoviridae), the family of lipophage (lipotricxividae), the family of polymorphic enveloped viruses (Plaolioviridae), the family of rod-forming phages (Rudiviidae), the genus salt terminal protein phages (Salterprovirus), and the family of biperiviridae (Bicaudaviridae).

In particular embodiments, the lytic bacteriophage is from a given bacterial species or strain. In another particular embodiment, the non-lytic bacteriophage is from the same or different given bacterial species or bacteria.

By "bacteriophage from a given bacterial species or strain" is meant herein a bacteriophage that specifically targets a particular bacterial species or strain and/or a bacteriophage that is hosted by a particular bacterial species or strain.

A non-exhaustive list of bacterial genera and their known host-specific bacterial viruses is presented in the following paragraphs. Synonyms and spelling variants are indicated in brackets. Homonyms repeat as frequently as they occur (e.g., D, D, d). Unnamed phages are indicated by "NN" beside their genus, and their numbers are given in brackets.

Bacteria of the genus actinomycetes (actinomycetes) can be infected with the following phages: av-I, av-2, av-3, BF307, CT1, CT2, CT3, CT4, CT6, CT7, CT8 and 1281.

Bacteria of the genus Aeromonas (Aeromonas) can be infected by the following phages: AA-I, aeh2, N, PMl, TP446, 3, 4, 11, 13, 29, 31, 32, 37, 43-10T, 51, 54, 55r.1, 56RR2, 57, 58, 59.1, 60, 63, aehl, F, PM2, 1, 25, 31, 40rr2.8t, (synonym=44R), (synonym=44rr2.8t), 65, PM3, PM4, PM5, and PM6.

Bacteria of the genus Bacillus (Bacillus) can be infected by the following phages: A. aizl, al-K-I, B, BCJAl, BCl, BC2, BLLl, BLl, BP, BSLl, BSL2, BSl, BS3, BS8, BS15, BS18, BS22, BS26, BS28, BS31, BS104, BS105, BS106, BTB, B1715V1, C, CK-I, coll, corl, CP-53, CS-I, CSi, D, D, D, D5, entl, FP8, FP9, FSi, FS2, FS3, FS5, FS8, FS9, G, GH, GT8, GV-I, GV-2, GT-4, g3, gl2, gl3, gl4, gl6, gl7, g21, g23, g24, g29, H2, kenl, KK-88, kuml, kyul, J W-1, LP52, (synonyms = LP-52), L7, mexl, MJ-I, mor2, MP-7, MPLO, MP12 MP14, MP15, neol, N2, N5, N6P, PBCl, PBLA, PBPl, P2, S-a, SF2, SF6, shal, sill, SP02, (synonym = ΦSPP1), SP beta, STI, STi, SU-Il, t, tbI, tb2, tb5, tbIO, tb26, tb51, tb53, tb55, tb77, tb97, tb99, tb560, tb595, td8, td6, tdl5, tgI, tg4, tg6, tg7, tg9, tgIO, tgIl, tgl, tgl5, tg21, tinl, tin7, tin8, tinl3, tm3, tocl, togl, toll, TP-I, TP-10vir, TP-15C, TP-16C, TP-17C, TP-19, TP35, TP51, TP-84, tt4, tt6, A, B, C, D, E, VA-9、W、wx23、wx26、Yunl、α、γ、pl l、/> IA. IB, 1-97A, 1-97B, 2, 3, 5, 12, 14, 20, 30, 35, 36, 37, 38, 41C, 51, 63, 64, 138D, I, II, IV, NN-Bacillus (13), alel, ARl, AR2, AR3, AR7, AR9, bace-11, (synonym=11), bastille, BLl, BL2, BL3, BL4, BL5, BL6, BL8, BL9, BP124, BS28, BS80, ch, CP-51, CP-54, D-5, darl, denl, DP-7, entl, foSi, foS2, FS4, FS6, FS7, G, gall, gamma, GEl, GF-2, GSi, GT-I, GT-2, GT-3, GT-4, GT-5, GT-6, GT-7, GV-6, gl5, 19 110, is, K, MP9, MP13, MP21, MP23, MP24, MP28, MP29, MP30, MP32, MP34, MP36, MP37, MP39, MP40, MP41, MP43, MP44, MP45, MP47, MP50, NLP-I, no. l, N17, N19, PBSl, PKl, PMBl, PMB, PMJl, S, SPOl, SP3, SP5, SP6, SP7, SP8, SP9, SPlO, SP-15, SP50, (synonym = SP-50), SP82, SST, subl, SW, tg, tgl2, tgl3, tgl4, thul, tha Λ, thas, tin4, tin23, TP-13, TP33, TP50, TSP-I, V type, VI type, V, vx, β22,> 1. 1, 2C, 3NT, 4, 5, 6, 7, 8, 9, 10, 12, 17, 18, 19, 21, 138, III, 4 (Bacillus megaterium), 4 (Bacillus sphaericus), AR13, BPP-IO, BS32, BS107, bl, B2, GA-I, GP-IO, GV-3, GV-5, g8, MP20, MP27, MP49, nf, PP5, PP6, SF5, tgl8, TP-I, versailles, ind >1-97、837/IV、/>Bacillus (1), butlo, BSLlO, BSLI, BS6, bsil, BS16, BS23, bsllol, BS102, gl8, morl, PBLl, SN, thu2, thu3, tmI, tm2, TP-20, TP21, TP52, F-type, G-type, IV-type, h N-BacMus (3), BLE, (synonym = oc), BS2, BS4, BS5, BS7, blO, B12, BS20, BS21, F, MJ-4, PBA12, AP50-04, AP50-11, AP50-23, AP50-26, AP50-27 and Bam35. The following bacillus-specific phages were defective: DLP10716, DLP-11946, DPB5, DPB12, DPB21, DPB22, DPB23, GA-2, M, no. IM, PBLB, PBSH, PBSV, PBSW, PBSX, PBSY, PBSZ, phi, spa, type 1 and μ.

Bacteria of the genus bacteroides can be infected with the following phages: crAss-phage, ad I2, baf-44, baf-48-B, baf-64, bf-I, bf-52, B40-8, fl, β1, 11. 67.1, 67.3, 68.1, mt-Bacteroides (3), bf42, bf71, bdellovibrio (Bdellovibrio) (1) and BF-41.

Bacteria of the genus Bordetella (Bordetella) can be infected with the following phages: 134 and NN-Botrytis (3).

Bacteria of the Borrelia genus (Borrelia) can be infected with the following phages: NN-borrelia (1) and NN-borrelia (2).

Bacteria of the genus Brucella (Brucella) can be infected with the following phages: a422, bk, (synonym=berkeley), BM29, FOi, (synonym= FOl), (synonym= FQl), D, FP2, (synonym=fp2), (synonym=fd 2), fz, (synonym=fz75/13), (synonym=firenze 75/13), (synonym=fi), fi, (synonym=fl), fim, (synonym= FIm), (synonym=fim), fiU, (synonym= FlU), (synonym= FiU), F2, (synonym=f2), F3, (synonym=f3), F4, (synonym=f4), F5, (synonym=f5), F6, F7, (synonym=f7), F25, (synonym=f25), F44, (synonym= -F44), F45U, (synonym=f25u), (synonym F48, F69, F48), (w=t35), (brm=t35), (f=t35), and (synonym=t35), (t=t35), and (synonym=t35) X, 3, 6, 7, 10/1, (synonym=10), (synonym=f8), 12m, 24/11, (synonym=24), (synonym=f9), 45/111, (synonym=45), 75, 84, 212/XV, (synonym=212), (synonym=fi 0), (synonym= FlO), 371/XXIX, (synonym=371), (syn=fn), (synonym=fl) and 513.

Bacteria of the genus Burkholderia (Burkholderia) can be infected with the following phages: CP75, NN-Burkholderia (1) and 42.

Bacteria of the genus Campylobacter (Campylobacter) can be infected with the following phages: type C, NTCC12669, NTCC12670, NTCC12671, NTCC12672, NTCC12673, NTCC12674, NTCC12675, NTCC12676, NTCC12677, NTCC12678, NTCC12679, NTCC12680, NTCC12681, NTCC12682, NTCC12683, NTCC12684, 32f, 111C, 191, NN-campylobacter (2), vfi-6, (synonym = V19), vfV-3, V2, V3, V8, V16, (synonym = Vfi-1), V19, V20 (V45), V45, (synonym = V-45) and NN-campylobacter (1).

Bacteria of the genus Chlamydia (Chlamydia) can be infected with the following phages: chpl.

Bacteria of the genus Clostridium (Clostridium) can be infected with the following phages: CAKl, CA5, CA7, CE beta, (synonym=1c), CE gamma, cldl, C-n71, C-203Tox-, de beta, (synonym=id), (synonym= lDt 0x+), HM3, KMl, KT, ms, NAl, (synonym=naltox+), PA135Oe, pf, PL73, PL78, PL81, PL, P50, P5771, P19402, lCt0x+, 2Ct0X 2d3 (synonym=2dt0x+), 3C, (synonym=3ctox+), 4C, (synonym=4ct0x+), 56, III-l, NN-Clostridium (61), NBlt0X+, alpha l, CAl, HMT, HM2, PFl 5P-23, P-46, Q-05, Q-Oe, Q-16, Q-21, Q-26, Q-40, Q-46, S111, SA02, WA01, WA03, wm, W523, 80, C, CA2, CA3, CPTl, CPT4, cl, C4, C5, HM7, H11/A1, H18/Ax, FWS23, hi58ZA1, K2ZA1, K21ZS23, ML, NA2t0X; pf2, pf3, pf4, S9ZS3, S41ZA1, S44ZS23, α2, 41, 112ZS23, 214/S23, 233/Ai, 234/S23, 235/S23, II-l, II-2, II-3, NN-Clostridium (12), CAl, fl, K, S2, 1, 5, and NN-Clostridium (8).

Bacteria of the genus Corynebacterium (Corynebacterium) can be infected with the following phages: CGKl (defective), A, A, A3, alOl, a128, a133, a137, a139, a155, a182, B, BF, B17, B18, B51, B271, B275, B276, B277, B279, B282, C, capi, CCl, CGl, CG2, CG33, CL31, cog, (synonym=cg 5), D, E, F, H, H-I, hqi, hq2, 11ZH33, ii/31, J, K, K, (synonym=ktox "), L, L, (synonym=ltox+), M, MC-I, MC-2, MC-3, MC-4, MLMa, N, O, ovi, ov2, ov3, P, P, R, RP, RS29, S, T, U, UB1, ub2, UH1, UH3, UH5, UH6, β, (synonym=βtox+), βhv64, βvir, γ, (synonym=γto-), γl9, δ, (δ= pto +), Φ -, (synonym), = pto-), and (n-)Omega, IA, 1/1180, 2/1180, 5ad/9717, 7/4465, 8ad/10269, 10/9253, 13Z9253, 15/3148, 21/9253, 28, 29, 55, 2747, 2893, 4498 and 5848.

Bacteria of the Enterococcus genus (Enterococcus) are infected with the following phages: DF78, fl, F2, 1, 2, 4, 14, 41, 867, dl, SB24, 2BV, 182, 225, C2F, E3, E62, DS96, H24, M35, P3, P9, SBlOl, S2, 2BII, 5, 182a, 705, 873, 881, 940, 1051, 1057, 21096C, NN-enterococcus (1), PEl, fl, F3, F4, VD13, 1, 200, 235 and 341.

Bacteria of the Erysipelothrix genus (erysiphe thrix) can be infected with the following phages: NN-erysipelothrix (1).

Bacteria of the genus Escherichia may be infected with the following phages: BW73, B278, D6, D108, E, el, E24, E41, FI-2, FI-4, FI-5, HI8A, ffl8B, i, MM, mu, (synonym = mu), (homonymMeaning = MuI), (synonym = Mu-I), (synonym = MuI), (synonym = μ), 025, phI-5, pk, PSP3, pl, plD, P2, P4 (defective), sl,(defective), -> Psi (defect type), 7A,15 (defective), 18, 28-1, 186, 299, H-Escherichia (2), AB48, CM, C4, C16, DD-VI, (synonym=dd-VI), (synonym=DDVI), (synonym=DDVi), E4, E7, E28, FIl, FI3, H, hl, H3, H8, K3, M, N, ND-2, ND-3, ND4, ND-5, ND6, ND-7, ox-I (synonym=OXl), (synonym=HF), ox-2 (synonym=0x2), ox-3, ox-4, ox-5, (synonym=0x5), ox-6, (synonym=66F), ox-4, ox-5,5 0111, phI-I, RB, RB43, RB49, RB69, S, saI-I, sal-2, sal-3, sal-4, sal-5, sal-6, TC23, TC45, tuII-6, (synonym=tuii), tuIP-24, tuII 46, tuIP-60, T2, (synonym=ganutia), (synonym=γ), (synonym=pc), (synonym=p.c.), (synonym=t-2), (synonym=t2), (synonym=p4), T4, (synonym=t4), T6, T35, αl, 1, IA, 3, (synonym=c3), 3a, 3t+, (synonym=3), (synonym=ml), and (synonym=ml) >9266Q, CFO, HK620, J, K, klF, m, 59, no. a, no. e, no.3, no.9, N4, sd, (synonym=sd), (synonym=sa) 3 (synonym=sd), a,(synonym=cd), T3, (synonym=t-3), (synonym=t3), T7, (synonym=t-7), (synonym=t7), WPK, W31, Δh,Φ04-CF、Φ05、Φ06、Φ07、Omega 8, 1, 3, 7, 8, 26, 27, 28-2, 29, 30, 31, 32, 38, 39, 42, 933W, NN-escherichia (1), esc-7-11, AC30, CVX-5, cl, DDUP, ECl, EC, E21, E29, fl, F26S, F S, hi, HK022, HK97, (synonym = Φhk 97), HK139, HK253, HK256, K7, ND-I, no. D, PA-2, q, S2, tl, (synonym = α), (synonym = P28), (synonym = T-I), (synonym = Tx), T3C, T, (synonym = T-5), (synonym = T5), UC-I, w, β4, γ2, λ (synonym = lambda), (synonym = λ), Φd326, -j, (synonym = P28)>Φ06、Φ7、Φ10、/>χ, (synonym=χi),2. 4, 4A, 6, 8A, 102, 150, 168, 174, 3000, AC6, AC7, AC28, AC43, AC50, AC57, AC81, AC95, HK243, klO, ZG/3A, 5A, 21EL, H19-J, 933H, O, 157 typing phage 1-16, JES-2013, 121Q, 172-1, 1720a-02, ADB-2, AKFV33, av-05, bV_EcoS_AHP42, bV_EcoS_AHP24, bC_EcoS_AHS24, bV_EcoS_AKS96, CBA120.

Bacteria of the genus fusobacterium are infected with the following phages: NN-Fusobacterium (2), fv83-554/3, fv88-531/2, 227, fv2377, fv2527 and fv8501.

Bacteria of the genus Haemophilus (Haemophilus) are infected with the following phages: HP1, S2 and N3.

Bacteria of the genus Helicobacter (Helicobacter) are infected with the following phages: HP1 and the genus-helicobacter (1).

Bacteria of the genus Klebsiella (Klebsiella) are infected with the following phages: AIO-2, KI4B, kl6B, kl9, (synonym=k19), kl14, kl15, kl21, kl28, kl29, KI32, kl33, kl35, kl106B, kl171B, kl181 6732B, AIO-I, AO-I, AO-2, AO-3, FC3-10, K, kl1, (synonym= KIl), kl2, (synonym=k12), kl3, (synonym=k13), (synonym=kl70/11), kl4, (synonym=k14), kl5, (synonym=k15), kl6, (synonym=k16), kl7, (synonym=k17), kl8, (synonym=k18), kl19, (synonym=k19), and so on Kl27, (synonym=k127), kl31, (synonym=k131), kl35, kl171 7513-I, kl4B, kl8, kl11, kl12, kl13, kl16, kl17, kl18, kl20, kl22, kl23, kl24, kl26, kl30, kl34, kl106B, KIi B, kl B, kl xi 328, P5046, 11, 380, III, IV, VII, VIII, FC3-11, kl2B, (synonym=k12B), kl25, (synonym=k125), kl42B, (synonym=k142) and (synonym=k142B, kl181B, (synonym= KIl 81) K1181B, kl765/! (synonym=k1765/1), kl842B (synonym=k1832B), kl937B (synonym=k1937b), ll, 7. 231, 483, 490, 632, and 864/100.

Bacteria of the genus leptospira (Lepitospira) are infected with the following phages: LEl, LE3, LE4 and-NN-leptospira (1).

Bacteria of the Listeria genus (Listeria) are infected with the following phages: a511, 01761, 4211, 4286, (synonym = BO 54), a005, a006, a020, a500, a502, a511, al 18, a620, a640, B012, B021, B024, B025, B035, B051, B053, B054, B055, B056, blOl, BI lO, B545, B604, B653, C707, D441, HSO47, hlOG, H8/73, H19, H21, H43, H46, H107, H108, HI lO, H163/84, H312, H340, H387, H391/73, H684/74, H924A, PSA, U153,(synonym = P35), 00241, 00611, 02971A, 02971C, 5/476, 5/911, 5/939, 5/11302, 5/11605, 5/11704, 184, 575, 633, 699/694, 744, 900, 1090, 1317, 1444, 1652, 1806, 1807, 1921/959, 1921/11367, 1921/11500, 1921/11566, 1921/12460, 1921/12582, 1967, 2389, 2425, 2671, 2685, 3274, 3550, 3551, 3552, 4276, 4277, 4292, 4477, 5337, 5348/11363, 5348/11646, 5348/12430, 5348/12434, 10072, 11355C, 11711A, 12029, 12981, 13441, 90666, 90816, 93253, 907515, 910716 and nnl-listeria (15).

Bacteria of the genus Morganella (Morganella) are infected with the following phages: 47.

bacteria of the genus Mycobacterium (Mycobacterium) are infected with the following phages: 13. AGl, ALi, ATCC 11759, A2, B.C3, BG2, BKl, BK5, butyl, B-I, B, B7, B30, B35, clark, cl, C2, DNAIII, DSP1, D4, D29, GS4E, (synonym=GS 4E), GS7, (synonym=GS-7), (synonym=GS 7), IPa, lacticola, legendre, leo, L5, (synonym=ΦL-5), MC-I, MC-3, MC-4, minetti, MTPHI, mx4, myF P/59a, phlei, (synonym=phlei1), phlei4, polonus II, rabinovitschi, smegmatis, TM4, TM9, TMlO, TM20, Y7, ylO,IB. IF, IH, 1/1, 67, 106, 1430, bl, (synonym=bol), B24, D, D29, F-K, F-S, HP, polonus I, roy, rl, (synonym=rl-Myb), (synonym=ri), 11, 31, 40, 50, 103a, 103B, 128, 3111-D, 3215-D, and NN-mycobacterium (1).

Bacteria of the Neisseria genus (neissenia) are infected with the following phages: group I, group II and NPl.

Bacteria of the genus Nocardia (Nocardia) are infected with the following phages: MNP8, NJ-L, NS-8, N5 and Ttin-nocardia.

Bacteria of the genus Proteus (Proteus) are infected with the following phages: pm5, 13vir, 2/44, 4/545, 6/1004, 13/807, 20/826, 57, 67b, 78, 107/69、121、9/0、22/608、30/680、PmI、Pm3、Pm4、Pm6、Pm7、Pm9、PmIO、PmI l、Pv2、πl、7/549、9B/2、10A/31、12/55、14、15、16/789、17/971、19A/653、23/532、25/909、26/219、27/953、32A/909、33/971、34/13、65、5006M、7480b、VI、13/3a、Clichy 12、π2600、/>1/1004, 5/742, 9, 12, 14, 22, 24/860, 2600/D52, pm8 and 24/2514.

Providencia (Providencia) bacteria are infected with the following phages: PL25, PL26, PL37, 9211/9295, 9213/921Ib, 9248, 7/R49, 7476/322, 7478/325, 7479, 7480, 9000/9402 and 9213/921Ia.

Bacteria of the genus Pseudomonas (Pseudomonas) are infected with the following phages: pfI, (synonym=pf-I), pf2, pf3, PP7, PRRl, 7S, im-pseudomonas (1), AI-I, AI-2, B17, B89, CB3, col 2, col 11, col 18, col 21, C154, C163, C167, C2121, E79, F8, ga, gb, H22, K1, M4, N2, nu, PB-I, (synonym=pbl), pfl6, PMN17, PPl, PP8, psal, psPl, psP2, psP3, psP4, psP5 PS3, PS17, PTB80, PX4, PX7, PYOl, PYO2, psP5, psP 6, psP 9, PYOlO, PYO13, psP 14, psP 16, psP 18, psP 19, psP 20, psP 29, psP 32, psP 33, psP 35, psP 36, psP 38, psP 39, psP 41, psP 42, psP 45, psP 47, psP 48, psP 64, psP 69, psP 103, psP2, S2, un-PsP 4, yan, (synonym = Φkz), - ->Φmu78、/> 1/72, 2/79, 3/DO, 4/237, 5/406, 6C, 6/6660, 7v, 7/184, 8/280, 9/95, 10/502, 11/DE, 12/100, 12S, 16, 21, 24, 25F, 27, 31, 44, 68, 71, 95, 109, 188, 337, 352, 1214, H N-Pseudomonas (23), A856, B26, CI-I, CI-2, C5, D, gh-1, fl 16, HF, H90, K5, K6, kl 04, K109 K166, K267, N4, N5, O6N-25P, PE, pf, PPN25, PPN35, PPN89, PPN91, PP2, PP3, PP4, PP6, PP7, PP8, PP56, PP87, PPl 14, PP206, PP207, PP306, PP651, psp231a, pssy401, pssy9220, psi, PTB2, PTB20, PTB42, PXL, PX3, PXLO, PX12, PX14, PYO, PYO71, R, SH6, SH133, tf, ya5, ya7, and @>ΦKf77、/>ΦmnF82、/>1. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12B, 13, 14, 15 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 31, 53, 73, 119x 145, 147, 170, 267, 284, 308, 525, NN-Pseudomonas (5), af, A7, B3, B33, B39, BI-I, C22, D3, D37, D40, D62, D3112, F7, flO, g, gd, ge, g ζ Hwl2, jb 19, KFL L °, OXN-32P, O6N-52P, PCH-I, PC-1, PC35-1, PH2, PH51, PH93, PH132, PMW, PM13, PM57, PM61, PM62, PM63, PM69, PM105, PMl, PM681, PM682, PO4, PPl, PP4, PP5, PP64, PP65, PP66, PP71, PP86, PP88, PP92, PP401, PP711, PP891, pssy41, pssy42, pssy403, pssy404, pssy420, pssy923, PS4, PS-IO, pz, SDl, SLl, SL3, SL5, SM, 2. 2F, 5, 7M, 11, 13/441, 14, 20, 24, 40, 45, 49, 61, 73, 148, 160, 198, 218, 222, 236, 242, 246, 249, 258, 269, 295, 297, 309, 318, 342, 350, 351, 357-1, 400-1, H, N-Pseudomonas (6), glol, M6a, ll, PB2, psyl 5, pssy4210, pssy4220, PYO12, PYO, PYO, PYO50, PYO, PYO, PYO53, PYO57, PYO, PYO, PX2, PX5, SL4, and so on>And 1214.

Bacteria of the genus Rickettsia (Rickettsia) are infected with the following phages: NN-rickettsia.

Bacteria of the genus Salmonella (Salmonella) are infected with the following phages: b. beccles, CT, d, dundee, f, feIs 2, GI, GUI, GVI, GVIII, k, K, I, j, L, 01, (synonym=0-1), (synonym=o1), (synonym=o-I), (synonym=7), 02, 03, P3, P9a, plO, sab3, sab5, sanlS, sanl7, SI, taunton, viI, (synonym= ViI), 9, imSalmonella (1), N-I, N-5, N-IO, N-17, N-22, 11, 12, 16-19, 20.2, 36, 449C/C178, 966A/C259, a, b.a.o.r., e, G4, GUI, L, LP7, M, MG, N-18, PSA68, P4, P9C, P22, (synonym=p22), (synonym=plt22), P22al, P22-4, P22-7, P22-11, SNT-I, SNT-2, SP6, villi, viIV, viV, viVI, viVII, worksop, sj, 34, 1, 37, and (40), (40=l) ])、1、422、2、2.5、3b、4、5、6、14(18)、8、14(6,7)、10、27、28B、30、31、32、33、34、36、37、39、1412、SNT-3、7-11、40.3、c、C236、C557、C625、C966N、g、GV、G5、Gl 73、h、IRA、Jersey、MB78、P22-1、P22-3、P22-12、Sabl、Sab2、Sab2、Sab4、Sanl、San2、San3、San4、San6、San7、San8、San9、Sanl3、Sanl4、Sanl6、Sanl8、Sanl9、San20、San21、San22、San23、San24、San25、San26、SasLl、SasL2、SasL3、SasL4、SasL5、SlBL、SII、ViII、1. 2, 3a, 3al, 1010, ym-Salmonella (1), N-4, sasL6 and 27.

Bacteria of the genus Serratia (Serratia) are infected with the following phages: A2P, PS, SMB3, SMP5, SM2, V40, V56, ic, ΦCP-3, ΦCP-6, 3M, 10/la, 20A, 34CC, 34H, 38T, 345G, 345P, 501B, SMB2, SMP2, BC, BT, CW2, CW3, CW4, CW5, lt232, L2232, L34, L.228, SLP, SMPA, V.43, sigma,Φcp6-1, Φcp6-2, Φcp6-5, 3T, 5, 8, 9F, 10/1, 2OE, 32/6, 34B, 34CT, 34P, 37, 41, 56D, 56P, 6OP, 61/6, 74/6, 76/4, 101/8900, 226, 227, 228, 229F, 286, 289, 290F, 512, 764a, 2847/10, 2847/1Oa, l.359, and SMBl.

Bacteria of the Shigella genus (Shigella) are infected with the following phages: fsa, (synonym=a), FSD2D, (synonym=d2d), (synonym=w2d), FSD2E, (synonym=w2e), fv, F6, F7.8, H-Sh, PE5, P90, sfII, sh, SHm, SHrv, (synonym=hiv), SHvi, (synonym=hvi), SHVvm, (synonym=hviii), skγ66, (synonym=γ66), (synonym=yββ), (synonym=γ6b), SKm, (synonym=siiib) 5 (synonym=ui), SKw, (synonym=sima), (synonym=iv), SIC ^TM (synonym=siva.), (synonym=iva), SKvi, (synonym= KVI), (synonym=svi), (synonym=vi), SKvm, (synonym=svm), (synonym=viii), SKV pi IA, (synonym=svma), (synonym=viiia), STvi, STK, STx1, STxn, S66, W2, (synonym=d2c), (synonym=d20), and,3-SO-R, 8368-SO-R, F, (synonym=fs7), (synonym=k29), flO, (synonym=fslo), and SO on,(synonym=k31), I1, (synonym=alfa), (synonym=fsa), (synonym=kl8), (synonym=α), I2, (synonym=a), (synonym=k19), SG33, (synonym=g35), (synonym=so-35/G), SG35, (synonym=so-55/G), SG3201, (synonym=so-3201/G), SHn, (synonym=hii), SHv, (synonym=shv), SHx, SHX, SKn, (synonym=k2), (synonym=kii), (synonym=sn), (synonym=ssii), (synonym=ii), SKrv, (synonym=sm), (synonym=iv), synonym (V), synonym=va, (synonym=swab), (synonym=iva), SKV, (synonym=k4), (synonym=kv), (synonym=sv), (synonym= SsV), (synonym=v), SKx, (synonym=k9), (sx= SsX), (SX) (synonym=x), STV, (synonym=t35), (synonym=35-50-R), STvm, (synonym=t8345), (synonym=8345-SO-S-R), W1, (synonym=d8), (synonym=fsd8), W2A, (synonym=d2a), (synonym=fs 2A), DD-2, sf6, FSi, (synonym=fl), sf6, (synonym=f6), SG42, (synonym=so-42/G), SG3203, (synonym=so-3203/G), SKF12, (synonym= SsF 12), (synonym=f12), STn, (synonym=1881-SO-R), γ66, (synonym=gamma 66 a), (synonym=ssγ66), >BIl, DDVII, (synonym=DD 7), FSD2B, (synonym=W2B), FS2, (synonym=F2), FS4, (synonym=F4), FS5, (synonym=F5), FS9, (synonym=F9), FIl, P2-S0-S, SG36, (synonym=SO-36/G), (synonym=G36), SG3204, and SO (synonym=so-3204/G), SG3244, (synonym=so-3244/G), SHi, (synonym=hi), SHv pi, (synonym=hvii), SHK, (synonym= HIX), SHx1, SHx pi, (synonym= HXn), SKI, KI, (synonym=s1), (synonym= SsI), SKVII, (synonym=kviii), (synonym=svpi), (synonym=ssvii), SKIX, (synonym= KIX), (synonym= HXnS1 x), (synonym=ssix), SKXII, (synonym=kxii), (synonym=sxn), (synonym=ssxii), STi, STffl, STrv, STVi, STv pi, S70, S206, U2-S0-S, 3210-SO-S, 3859-SO-S, 4020-SO-S, SHm, (synonym=hpi i), SH χi, (synonym= HXt) and SKxI, (synonym= KXI), (synonym=sχi), (synonym=ssxi), (synonym=xi).

Bacteria of the genus Staphylococcus (Staphylococcus) are infected with the following phages: A. EW, K, ph5, ph9, phIO, phl3, pl, P2, P3, P4, P8, P9, plO, RG, SB-I, (synonym = Sb-I), S3K, twort, Φsk311, 06. 40, 58, 119, 130, 131, 200, 1623, STCl, (synonym = STCl), STC2, (synonym = STC 2), 44AHJD, 68, ACl, AC2, A6"C", A9"C", b581, CA-I, CA-2, CA-3, CA-4, CA-5, DI L, L39X35, L54a, M42, nl, N2, N3, N4, N5, N7, N8, nlO, ni L, N12, N13, N14, N16, ph6, phl2, phl4, UC-18, U4, U15, sl, S2, S3, S4, S5, X2, Z1>ω、11、/> (synonym = P11-M15), 15, 28A, 29, 31B, 37, 42D, (synonym = P42D), 44A, 48, 51, 52A, (synonym = P52A), 52B, 53, 55, 69, 71, (synonym = P71), 71A, 72, 75, 76, 77, 79, 80 a, 82A, 83A, 84, 85, 86, 88A, 89, 90, 92, 95, 96, 102, 107,108、111、129-26、130、130A、155、157、157A、165、187、275、275A、275B、356、456、459、471、471A、489、581、676、898、1139、1154A、1259、1314、1380、1405、1563、2148、2638A、2638B、2638C、2731、2792A、2792B、2818、2835、2848A、3619、5841、12100、AC3、A8、AlO、A13、b594n、D、HK2、N9、N15、P52、P87、Sl、S6、Z4、/>3A, 3B, 3C, 6, 7, 16, 21, 42B, 42C, 42E, 44, 47a5 47C, 51, 54x1, 70, 73, 75, 78, 81, 82, 88, 93, 94, 101, 105, 110, 115, 129/16, 174, 594n, 1363/14, 2460 and mS-staphylococcus (1).

Bacteria of the streptococcus genus are infected with the following phages: EJ-I, NN-Streptococcus (1), a, cl, FL0Ths, H39, cp-I, C ρ -5, cp-7, cp-9, cp-IO, AT298, A5, alO/Jl, alO/J2, alO/J5, alO/J9, A25, BTI, b6, CAl, c20-l, c20-2, DP-I, dp-4, DTl, ET42, elO, FA101, FEThs, fkappa, FKKIOI, FKLIO, FKP, FKH, FLOThs, fyIOl, FL, F, F20140/76, g, GT-234, HB3, (synonym = HB-3), HB-623, HB-746, M102, O1205), PST, PO, pl, P2, P3, P5, P6, P8, P9, P12, P13, P14, P49, P50, P51, P52, P53, P54, P55, P56, P57, P58, P59, P64, P67, P69, P71, P73, P75, P76, P77, P82, P83, P88, sc, sch, sf, sfIl1, (synonym = SFiI l),>(synonym = Φsfil), SFil9, (synonym=sfil9), -j>Sfi21, (synonym=sfi21), -j>ST0、STX、st2、ST2、ST4、S3、/>s265、Φ17、/>Φ57、/> Φ7201, ωl, ω2, ω3, ω4, ω5, ω6, ω8, ω lO, 1, 6, 9, 1OF, 12/12, 14, 17SR, 19S, 24, 50/33, 50/34, 55/14, 55/15, 70/35, 70/36, 71/ST15, 71/45, 71/46, 74F, 79/37, 79/38, 80/J4, 80/J9, 80/ST16, 80/15, 80/47, 80/48, 101, 103/39, 103/40, 121/41, 121/42, 123/43, 123/44, 124/44, 337/ST17 and mStreptococcus (34).

Bacteria of the Treponema genus (Treponema) are infected with the following phages: NN-Treponema (1).

Bacteria of the genus Vibrio (Vibrio) are infected with the following phages: CTX- 1、>HC2-2、ΦHC2-3、ΦHC2-4、ΦHC3-1、ΦHC3-2、ΦHC3-3、ΦHD1S-1、ΦHD1S-2、ΦHD2S-1、ΦHD2S-2、ΦHD2S-3、ΦHD2S-4、ΦHD2S-5、ΦHDO-1、ΦHDO-2、ΦHDO-3、ΦHDO-4、ΦHDO-5、ΦHDO-6、ΦKL-33、ΦKL-34、ΦKL-35、ΦKL-36、ΦKWH-2、ΦKWH-3、ΦKWH-4、ΦMARQ-1、ΦMARQ-2、ΦMARQ-3、ΦMOAT-1、ΦO139、ΦPEL1A-1、ΦPEL1A-2、ΦPEL8A-1、ΦPEL8A-2、ΦPEL8A-3、ΦPEL8C-1、ΦPEL8C-2、ΦPEL13A-1、ΦPEL13B-1、ΦPEL13B-2、ΦPEL13B-3、ΦPEL13B-4、ΦPEL13B-5、ΦPEL13B-6、ΦPEL13B-7、ΦPEL13B-8、ΦPEL13B-9、ΦPEL13B-10、 1-II, 5, 13, 14, 16, 24, 32, 493, 6214, 7050, 7227, II, (synonym = group II), -a-j->V, VIII, -m-Vibrio (13), KVP20, KVP40, nt-1, O6N-22P, P, el, e2, e3, e4, e5, FK, G, I, K, nt-6, nl, N2, N3, N4, N5, O6N-34N-P, OXN-72-P, OXN-85P, OXN-100P, P, ph-I, PL163/10, Q, S, T,>1-9, 37, 51, 57, 70A-8, 72A-4, 72A-10, 110A-4, 333, 4996, I (synonym=I group), III (synonym=III group), VI (synonym=A-Sarkov), VII, IX, X, H N-Vibrio (6), pAl, 7-8, 70A-2, 71A-6, 72A-5, 72A-8, 108A-10, 109A-6, 109A-8, lOA-1, 110A-5, 110A-7, hv-1, OXN-52P, P, P38, P53, P65, P108, picl, TPl3 VP3, VP6, VP12, VP13, 70A-3, 70A-4, 70A-10, 72A-1, 108A-3, 109-B1, 110A-2, 149>IV, (synonym = group IV), NN-vibrio (22), VP5, VPIl, VP15, VP16,αl, α2, α3a, α3b, 353B and H N-Vibrio (7).

Bacteria of the genus Yersinia (Yersinia) are infected with the following phages: H. H-I, H-2, H-3, H-4, lucas110, lucas 303, lucas 404, yerA3, yerA7, yerA20, yerA41, 3/M64-76, 5/G394-76, 6/C753-76, 8/C239-76, 9/F18167, 1701, 1710, PST, 1/F2852-76, D' Herelle, EV, H, kotljarova, PTB, R, Y, yerA, and, 12, 3, 4/C1324-76, 7/F783-76, 903, 1/M6176 and Yer2AT.

In a particular embodiment, the lytic bacteriophage is selected from the lytic bacteriophages listed above, and the non-lytic bacteriophage is selected from the non-lytic bacteriophages listed above.

In the context of a particular embodiment of the present invention, the lytic bacteriophage and/or non-lytic bacteriophage is selected from Salmonella virus SKML39, shigella virus AG3, dickeya virus (Dickeya) virus Limestone, dickeya virus RC2014, escherichia virus CBA120, escherichia virus PhaxI, salmonella virus 38, salmonella Det7, salmonella virus GG32, salmonella virus PM10, salmonella virus SFP10, salmonella virus SH19, salmonella virus SJ3, escherichia virus ECML4, salmonella virus Marshall, salmonella virus Maynard, salmonella virus SJ2, salmonella virus STML131, salmonella virus ViI, erwinia virus Ea2809, klebsiella virus 0507KN21, serratia virus IME250, serratia virus MAM1, campylobacter virus CP21, campylobacter virus CP220, salmonella virus Campylobacter virus CPt10, campylobacter virus IBB35, campylobacter virus CP81, campylobacter virus CP30A, campylobacter virus CPX, campylobacter virus NCTC12673, erwinia virus Ea214, erwinia virus M7, escherichia virus AYO A, escherichia virus EC6, escherichia virus HY02, escherichia virus JH2, escherichia virus TP1, escherichia virus VpaE1, escherichia virus wV8, salmonella virus FelixO1, salmonella virus HB2014, salmonella virus Mushroum, salmonella virus UAB87, citrobacter (Citrobacter) virus Mogle, citrobacter virus Mordin, escherichia virus SUSP1, escherichia virus HP2, aeromonas virus phiO18P, haemophilus virus 1, haemophilus virus 2, haemophilus virus HP2, salmonella virus, pasteurella (Pasteurella) virus F108, vibrio virus K139, vibrio virus Kappa, burkholderia virus phi52237, burkholderia virus phiE122, burkholderia virus phiE202, escherichia virus 186, escherichia virus P4, escherichia virus P2, escherichia virus Wphi, mannheimia (Mannheimia) virus PHL101, pseudomonas virus phiCTX, ralstonia (Ralstonia) virus RSA1, salmonella virus Fels2, salmonella virus PsP, salmonella virus SopEphi, yersinia virus L413C, staphylococcus virus G1, staphylococcus virus G15, staphylococcus virus 7, staphylococcus virus K, staphylococcus virus MCE2014, staphylococcus virus P108, staphylococcus virus S253, staphylococcus virus S25, staphylococcus virus S25, and Staphylococcus 4 Staphylococcus virus SA12, listeria virus A511, listeria virus P100, staphylococcus virus Remus, staphylococcus virus SA11, staphylococcus virus Stau2, bacillus virus Camphawk, bacillus virus SPO1, bacillus virus BCP78, bacillus virus TsirBombab, staphylococcus virus Twit, enterococcus virus phiEC24C, lactobacillus (Lactobacillus) virus Lb338-1, lactobacillus virus LP65, enterobacter virus PG7, escherichia virus CC31, klebsiella virus JD18, klebsiella virus PKO111, escherichia virus Bp7, escherichia virus IME08, escherichia virus JS10, escherichia virus JS98, escherichia virus QL01, escherichia virus VR5, enterobacter virus Eap3, klebsiella virus KP15, klebsiella virus KP27, klebsiella virus Matisse, klebsiella virus Miro, citrobacter virus Merlin, citrobacter virus Moon, escherichia virus JSE, escherichia virus phi1, escherichia virus RB49, escherichia virus HX01, escherichia virus JS09, escherichia virus RB69, shigella virus UTAM, salmonella virus S16, salmonella virus STML198, vibrio virus KVP40, vibrio virus nt1, vibrio virus ValKK3, escherichia virus VR7, escherichia virus VR20, escherichia virus VR25, escherichia virus VR26, shigella virus SP18, escherichia virus AR1, escherichia virus C40, escherichia virus E112, escherichia virus HY 134, escherichia virus HY01, escherichia virus E09, im 3, im 09 Escherichia virus RB14, escherichia virus T4, shigella virus Pss1, shigella virus Shfl2, yersinia virus D1, yersinia virus PST, acinetobacter (Acinetobacter) virus 133, aeromonas virus 65, aeromonas virus Aeh1, escherichia virus RB16, escherichia virus RB32, escherichia virus RB43, pseudomonas virus 42, cronobacter (Cronobacter) virus CR3, cronobacter virus CR8, cronobacter virus CR9, cronobacter virus PBES02, pectobacterium (Pectobacter) virus phiTE, cronobacter virus GAP31, escherichia virus 4MG, salmonella virus SSE121, escherichia virus FFH2, escherichia virus FV3, JEV 5, salmonella virus, brevibacterium (Brevibacillus) virus Abouo, brevibacillus virus Davies, bacillus Agate, bacillus virus Bobb, bacillus virus Bp8pC, erwinia virus Deimos, erwinia virus Ea35-70, erwinia virus RAY, erwinia virus Simmy50, erwinia virus specialty G, acinetobacter virus AB1, acinetobacter virus AB2, acinetobacter virus AbC62, acinetobacter virus AP22, arthrobacter (Arthrobacter) virus ArV, arthrobacter virus Trina, bacillus virus AvesoBmore, bacillus virus B4, bacillus virus Bigbertha, bacillus virus Riley, bacillus virus Spock, bacillus virus Troll, bacillus virus Bastille, bacillus virus 003, bacillus virus Bc 82, bc 1, bc431 virus Bacillus virus BM15, bacillus virus deep, bacillus virus JBP901, burkholderia virus Bsep 1, burkholderia virus Bsep 43, burkholderia virus Bsep 781, burkholderia virus BsepNY 3, xanthomonas (Xanthomonas) virus OP2, burkholderia virus Bsepmu, burkholderia virus phiE255, aeromonas virus 44RR2, mycobacterium virus Alice, mycobacterium virus Bxz, mycobacterium virus Dandelion, mycobacterium virus HyRo, mycobacterium virus I3, mycobacterium virus Nappy, mycobacterium virus Sebata, clostridium virus phiC2, clostridium virus phiCD27, clostridium virus phiCD119, bacillus virus CP51, bacillus virus JL, bacillus, escherichia virus M10, and Bacillus virus M10, erwinia virus Asesino, erwinia virus EaH2, pseudomonas virus EL, halomonas (Halomonas) HAP1, vibrio virus VP882, brevibacterium virus Jimmer, brevibacterium virus Osiris, pseudomonas virus Ab03, pseudomonas virus KPP10, pseudomonas virus PAKP3, sinorhizobium (Sinorhizobium) virus M7, sinorhizobium virus M12, sinorhizobium virus N3, erwinia virus Machina, arthrobacter virus Brent, arthrobacter virus Jawnski, arthrobacter virus Martha, arthrobacter virus Sonny, edwardsiella virus MSW3, edwardsiella virus PEi21, escherichia Mucor, shigella virus Sph, salmonella virus (Halerium) spore, bacillus sp.1, bacillus sp.sp.1, and E.sp.sp.1 Aeromonas virus 43, escherichia virus P1, pseudomonas virus CAb02, pseudomonas virus JG004, pseudomonas virus PAKP1, pseudomonas virus PAKP4, pseudomonas virus PaP1, burkholderia virus BsepF 1, pseudomonas virus 141, pseudomonas virus Ab28, pseudomonas virus DL60, pseudomonas virus DL68, pseudomonas virus F8, pseudomonas virus Pseudomonas virus JG024, pseudomonas virus KPP12, pseudomonas virus LBL3, pseudomonas virus LMA2, pseudomonas virus PB1, pseudomonas virus SN, pseudomonas virus PA7, pseudomonas virus phiKZ, rhizobium (Rhizobium) virus RHEPH4, rockwell virus RSF1, rockwell virus RSL2, rockwell virus RSL1, aeromonas virus 25, aeromonas virus 31, aeromonas virus Aes12, aeromonas virus Aes508, aeromonas virus AS4, aeromonas (stenotophomonas) virus IME13, staphylococcus virus IPLAC1C, staphylococcus virus SEP1, salmonella virus SPN3US, bacillus virus 1, geobacillus (Geobacillus) virus GBSV1, yersinia virus R1RT, yersinia virus TG1, bacillus virus G, bacillus virus PBS1, microcystis (Microcystis) virus Ma-LMM01, vibrio virus MAR, vibrio virus VHML, vibrio virus VP585, bacillus virus 13, bacillus virus Hakuna, bacillus virus Megann, bacillus virus WPh, acinetobacter virus AB3, acinetobacter virus p1, acinetobacter virus Fr1, acinetobacter 200 Acinetobacter virus PD6A3, acinetobacter virus PDAB9, acinetobacter virus phiAB1, escherichia virus K30, klebsiella virus K5, klebsiella virus K11, klebsiella virus Kp1, klebsiella virus KP32, klebsiella virus KpV289, klebsiella virus F19, klebsiella virus K244, klebsiella virus Kp2, klebsiella virus KP34, klebsiella virus Klebsiella virus KpV, klebsiella virus 3271, klebsiella virus KpV475, klebsiella virus SU503, klebsiella virus SU552A, pantoea (Pantoea) virus Limelight, pantoea virus Limezero, pseudomonas virus LKA1, pseudomonas virus phiKMV, xanthomonas virus F20, xanthomonas virus F30, murilla (Xylella) virus Prado, erwinia virus Era, erwinia virus, escherichia virus K5, escherichia virus K1-5, escherichia virus K1E, salmonella virus SP6, escherichia virus T7, kluyvera (Kluyvera) virus Kvp1, pseudomonas virus gh1, prochlorococcus (Prochlorococcus) virus PSSP7, synechococcus (Synechococcus) virus P60, syn5, streptococcus virus Cp1, streptococcus virus Cp7, staphylococcus virus 44AHJD, streptococcus virus C1, bacillus virus B103, bacillus virus GA1, bacillus virus phi29, kurthia (Kurthia) virus 6, actinomyces virus 1, mycoplasma virus P1, escherichia virus 24B, escherichia virus 933W, escherichia virus Min27, escherichia virus 28, st 2, st 02J 75J 13, shigella virus II, shigella virus 75J 13, and so on. Escherichia virus 191, escherichia virus PA2, escherichia virus TL2011, shigella virus VASD, burkholderia virus Bsep 22, burkholderia virus Bsepil 02, burkholderia virus Bsepmigl, burkholderia virus DC1, bodhami virus BPP1, burkholderia virus BsepC 6B, cellularum (Cellulophaga) virus Cba41, cellophaga virus Cba172, goldkola (Dinonoseobabacter) virus DFL12, erwinia virus Ea9-2, erwinia virus Frozen, escherichia virus phiV10, salmonella virus Epsilon15, salmonella virus SPN1S, pseudomonas virus F116, pseudomonas virus H66, escherichia AP5, APE.EC 4, and Erwinia virus EC 4, escherichia virus EC1UPM, escherichia virus ECBP1, escherichia virus G7C, escherichia virus IME11, shigella virus Sb1, achromobacter (Achromobacter) virus Axp3, achromobacter virus JWAlpha, edwardsiella virus KF1, pseudomonas virus KPP25, pseudomonas virus R18, pseudomonas virus Ab09, pseudomonas virus LIT1, pseudomonas virus PA26, pseudomonas virus LIT1, and Pseudomonas Ab22, pseudomonas CHU, pseudomonas LUZ24, pseudomonas PAA2, pseudomonas PaP3, pseudomonas PaP4, pseudomonas TL, pseudomonas KPP21, pseudomonas LUZ7, escherichia N4, salmonella 9NA, salmonella SP069, salmonella BTP1, salmonella HK620 Salmonella virus P22, salmonella virus ST64T, shigella virus Sf6, bacillus virus Page, bacillus virus Palmer, bacillus virus Pascal, bacillus virus Pony, bacillus virus Pookie, escherichia virus 172-1, escherichia virus ECB2, escherichia virus NJ01, escherichia virus phiEco32, escherichia virus Septima11, escherichia virus SU10, brula virus Pr, brula virus Tb, escherichia virus Pollock, salmonella virus FSL-058, salmonella virus FSL-SP-076, helicobacter virus 1961P, helicobacter virus KHP30, helicobacter virus KHP40, hamiltonia (Hamiltonia) virus APSE1, lactococcus (KSLacca) virus Y1, WMUlmi 3, WMUlmi 4, WMUlmi 3, and Ypsilosis, pseudomonas virus 119X, roselle (Roseobacillus) virus SIO1, vibrio virus VpV, vibrio virus VC8, vibrio virus VP2, vibrio virus VP5, streptomyces (Streptomyces) virus Amela, streptomyces virus phiCAM, streptomyces virus Aarocola, streptomyces virus Caliburn, streptomyces virus Danzina, streptomyces virus hydroa, streptomyces virus Izzy, streptomyces virus Lannister, streptomyces virus Lika Streptomyces virus Sujidade, streptomyces virus Zemlya, streptomyces virus ELB20, streptomyces virus R4, streptomyces virus phiHau3, mycobacterium virus Acandian, mycobacterium virus Baee, mycobacterium virus Reprobabate, mycobacterium virus Adawi, mycobacterium virus Bane1, mycobacterium virus BrownCNA, mycobacterium virus Chrisnmic, mycobacterium virus Cooper, mycobacterium virus JAMaL, mycobacterium virus Nigel Mycobacterium virus Stinger, mycobacterium virus Vincenzo, mycobacterium virus Zemanar, mycobacterium virus Apizium, mycobacterium virus Manad, mycobacterium virus Oline, mycobacterium virus Osmaximus, mycobacterium virus Pg1, mycobacterium virus Soto, mycobacterium virus Suffolk, mycobacterium virus Athena, mycobacterium virus Bernardo, mycobacterium virus Gadjet, mycobacterium virus Pipefish, mycobacterium virus Godines, mycobacterium virus Rosebus, mycobacterium virus Babsiella, mycobacterium virus Brujita, mycobacterium virus Che9c, mycobacterium virus Sbash, mycobacterium virus Hawkeye, mycobacterium virus Plot, salmonella virus AG11, salmonella virus Ent1, salmonella virus 18SE, salmonella virus Jey, salmonella virus L13, salmonella virus LS1 PA, salmonella virus SE2, salmonella virus SETP3, salmonella virus SETP7, salmonella virus SETP13, salmonella virus LN 101, salmonella virus SS3e, salmonella virus wksl3, escherichia virus K1G, escherichia virus K1H, escherichia virus K1ind1, escherichia virus K1ind2, salmonella virus SP31, leuconostoc (Leuconostoc) virus Lmd1, leuconostoc virus LN03, leuconostoc virus LN04, leuconostoc virus LN12, leuconostoc virus LN6B, leuconostoc virus P793, leuconostoc virus 1A4, leuconostoc virus Ln8, leuconostoc virus Ln9, leuconostoc virus LN25, leuconostoc virus LN34, leuconostoc virus TR3, bacillus virus Bong, bacillus Reng, mycobacterium, miche, mycobacterium and Mycobacterium Mycobacterium virus Charlie, mycobacterium virus Picqueaks, mycobacterium virus Xeno, mycobacterium virus Panchino, mycobacterium virus Phrann, mycobacterium virus Redi, mycobacterium virus Skinyp, gordonia virus BaxterFox, gordonia virus Yezy, gordonia virus Kita, gordonia virus Zirinka, gordonia virus Nymphara, mycobacterium virus Bignuz, mycobacterium virus Brusacram, mycobacterium virus Donovan, mycobacterium virus hburn, mycobacterium virus Jebeks, mycobacterium virus Malihi, mycobacterium virus yonce, enterobacter virus F20, klebsiella virus 1513, klebsiella virus KLPN1, klebsiella virus KP36, klebsiella virus P24, klebsiella AH, klebsiella 24, and so called after the use of the virus, escherichia virus AKS96, escherichia virus C119, escherichia virus E41C, escherichia virus Eb49, escherichia virus Jk06, escherichia virus KP26, escherichia virus Rogue1, escherichia virus ACGM12, escherichia virus Rtp, escherichia virus ADB2, escherichia virus JMW 1, escherichia virus JMW 2, escherichia virus T1, shigella virus PSf2, shigella virus Shigella virus Shfl1, citrobacter virus Stevie, escherichia virus TLS, salmonella virus SP126, cronobacter virus Esp2949-1, pseudomonas virus Ab18, pseudomonas virus Ab19, pseudomonas virus PaMx11, arthrobacter virus Amigo, propionibacterium virus Anatole, propionibacterium virus B3, bacillus virus Andromada, bacillus virus Blastoid, bacillus virus Curly, bacillus virus Eoghan, bacillus virus Finn, bacillus virus Glitterin, bacillus virus Riggi, bacillus virus Taylor, gordon virus Attis, mycobacterium virus Barnyard, mycobacterium virus Konstantinine, mycobacterium virus Predator, mycobacterium virus Bernal13, staphylococcus virus 77, staphylococcus virus 108PVL, mycobacterium virus Bron, mycobacterium virus Faith1, mycobacterium virus Joedirt, mycobacterium virus Rumpelstin, lactobacillus virus bIL67, lactobacillus virus C2, lactobacillus virus C5, lactobacillus virus Ld3, lactobacillus virus Ld17, lactobacillus virus Ld25A, lactobacillus virus LLdR, lactobacillus virus phLdb, cellophagostomum virus C121, cellophagostomum virus Cbavirus Cb 171, and Phacophagostomum virus 181, bacillus virus 250, bacillus virus IEBH, mycobacterium virus Ardmere, mycobacterium virus Avani, mycobacterium virus Boomer, mycobacterium virus Che8, mycobacterium virus Che9D, mycobacterium virus Deadp, mycobacterium virus Dlane, mycobacterium virus Dorothy, mycobacterium virus Dotproduct, mycobacterium virus Drago, mycobacterium virus Frutroop, mycobacterium virus Gumbie, mycobacterium virus Ibhube, mycobacterium virus Llij, mycobacterium virus Mozy, mycobacterium virus Mutaformia 13, mycobacterium virus Pacc40, mycobacterium virus PMC, mycobacterium virus Raey, mycobacterium virus Rockyrror, mycobacterium virus SG4, mycobacterium virus Shauna1, mycobacterium virus Shacan, mycobacterium virus Sparus, mycobacterium virus Taj, mycobacterium Wee, mycobacterium Mycobacterium virus Yoshi, salmonella virus Chi, salmonella virus FSLSP030, salmonella virus FSLSP088, salmonella virus iEPS5, salmonella virus SPN19, mycobacterium virus 244, mycobacterium virus Bask21, mycobacterium virus CJW1, mycobacterium virus Eureka, mycobacterium virus Kostya, mycobacterium virus Porky, mycobacterium virus Pumpkin, mycobacterium virus Sirduracell, mycobacterium virus Toto, mycobacterium virus Corndog, mycobacterium virus Firecracker, rhodobacter (Rhodobater) virus RcCroneus, pseudomonas virus D3112, pseudomonas virus DMS3, pseudomonas virus FHA0480, pseudomonas virus LPB1, pseudomonas virus MP22, pseudomonas virus MP29, pseudomonas virus MP38, pseudomonas virus KOR, pseudomonas virus D3, pseudomonas virus G1, pseudomonas virus, the genus Arthrobacter virus Decurro, the genus Gordomorphs Katyusha, the genus Gordomorphs Kvothes, the genus Propionibacterium virus B22, the genus Propionibacterium Doucete, the genus Propionibacterium virus E6, the genus Propionibacterium virus G4, the genus Burkholderia virus phi6442, the genus Burkholderia virus phi1026B, the genus Burkholderia virus phiE125, the genus Edwardsiella virus eiAU, the genus Mycobacterium virus Ff47, the genus Mycobacterium virus Muddy, the genus Mycobacterium virus Gaia, the genus Mycobacterium virus Giles, the genus Arthrobacter virus Captenmurica, the genus Arthrobacter Gordon, the genus Gordomorphs, the genus Gordon 2, the genus Paenium (Paenibacillus) Harrison, the genus EK99P1, the genus Escherichia virus HK, the genus JL 578, the genus Soddalike 1, the genus Edison 1, the genus Ala, the genus Aldribba, the genus Aldrips 2008 virus YD 1, the genus Aldrips (YD 1) Escherichia virus HK022, escherichia virus HK75, escherichia virus HK97, escherichia virus HK106, escherichia virus HK446, escherichia virus HK542, escherichia virus HK544, escherichia virus HK633, escherichia virus mEp234, escherichia virus mEp, escherichia virus mEpX1, escherichia virus mEpX2, escherichia virus mEp043, escherichia virus 35213 Escherichia virus mEp237, escherichia virus mEp, escherichia virus mEp460, escherichia virus mEp505, escherichia virus mEp, brevibacterium virus Jent, achromobacter virus 83-24, achromobacter virus JWX, arthrobacter virus Kellezzio, arthrobacter virus Kitkat, arthrobacter virus Bennie, arthrobacter virus DrRobert, arthrobacter virus Glenn, arthrobacter virus HunterDalle, arthrobacter virus, the genus Arthrobacter virus Joann, arthrobacter virus Korra, arthrobacter virus Preamble, arthrobacter virus Pumancar, arthrobacter virus Wayne, mycobacterium virus Alma, mycobacterium virus Arturo, mycobacterium virus Astro, mycobacterium virus Backyardagan, mycobacterium virus BBPiebs31, mycobacterium virus Benedict, mycobacterium virus Bethlehem, mycobacterium virus Billknuckles, mycobacterium virus Brns, mycobacterium virus Bxb1, mycobacterium virus Bxz2, mycobacterium virus Che12, mycobacterium virus Cuco, mycobacterium virus D29, mycobacterium virus Doom, mycobacterium virus icb, mycobacterium virus Ehoria, mycobacterium virus George, mycobacterium virus Glader, mycobacterium virus Goose, mycobacterium virus Hammer, heldavirus, mycobacterium virus Jaer, mycobacterium virus JC27 Mycobacterium virus Jeffibunny, mycobacterium virus JHC117, mycobacterium virus KBG, mycobacterium virus Kssjeb, mycobacterium virus Kugel, mycobacterium virus L5, mycobacterium virus Lesedi, mycobacterium virus LHTSCC, mycobacterium virus lockley, mycobacterium virus Marcel, mycobacterium virus Microwolf, mycobacterium virus Mrgordo, mycobacterium virus Museum, mycobacterium virus Nepal, mycobacterium virus Packman, mycobacterium virus Peaches, mycobacterium virus Perseus, mycobacterium virus Pukovnik, mycobacterium virus Rebeuca, mycobacterium virus Redr, mycobacterium virus Ridgecb, mycobacterium virus Rockstar, mycobacterium virus Saintus, mycobacterium virus Skipe, mycobacterium virus on, mycobacterium virus Itzer, mycobacterium virus SWU1, mycobacterium virus Pyger 17a, mycobacterium Swinga, mycobacterium Swingta, mycobacterium virus Timshel, mycobacterium virus Trixie, mycobacterium virus Turbo, mycobacterium virus Twister, mycobacterium virus U2, mycobacterium virus Violet, mycobacterium virus Wonder, escherichia virus DE3, escherichia virus HK629, escherichia virus HK630, escherichia virus Lambda, arthrobacter virus Laroye, mycobacterium virus Halo, mycobacterium virus Lieffie, mycobacterium virus Marvin, mycobacterium virus Mosmoris, arthrobacter virus Cium, arthrobacter virus Mudcat, escherichia virus N15, escherichia virus 9g, escherichia virus JenK1, escherichia virus JenP2, pseudomonas virus NP1, pseudomonas virus Pax25, bakan virus, mycobacterium virus Omujou, mycobacterium virus Cotta, mycobacterium virus Mycobacterium virus Optimus, mycobacterium virus Thibault, polarobacter (Polarobacter) virus P12002L, polarobacter virus P12002S, non-slip bacterium (Nonlabens) virus P12024L, non-slip bacterium virus P12024S, thermus (Thermus) virus P23-45, thermus virus P74-26, listeria virus LP37, listeria virus LP110, listeria virus LP114, listeria virus P70, propionibacterium virus ATCC29399BC, propionibacterium virus ATCC29399BT, propionibacterium virus atacne, propionibacterium virus Keiki, propionibacterium virus Kubed, propionibacterium virus Lauchelly, propionibacterium virus MK, propionibacterium virus Ouroxos, propionibacterium virus P91, propionibacterium virus P105, propionibacterium virus P1001, propionibacterium virus P1. Propionibacterium virus, propionibacterium virus P100A, propionibacterium virus P100D, propionibacterium virus P101A, propionibacterium virus P104A, propionibacterium virus PA6, propionibacterium virus Pacnes201215, propionibacterium virus PAD20, propionibacterium virus PAS50, propionibacterium virus PHL009M11, propionibacterium virus PHL025M00, propionibacterium virus PHL037M02, propionibacterium virus PHL041M10, propionibacterium virus PHL060L00, propionibacterium virus PHL067M01, propionibacterium virus PHL070N00, propionibacterium virus PHL071N05, propionibacterium virus PHL082M03, propionibacterium virus PHL092M00, propionibacterium virus PHL095N00, propionibacterium virus PHL111M01, propionibacterium virus PHL112N00, propionibacterium PHL113M01, propionibacterium virus PHL 114M 00, propionibacterium virus PHL116, propionibacterium virus PHL00, PHL 01N 01, PHL132 and PHL132 Propionibacterium virus PHL151M00, propionibacterium virus PHL151N00, propionibacterium virus PHL152M00, propionibacterium virus PHL163M00, propionibacterium virus PHL171M01, propionibacterium virus PHL179M00, propionibacterium virus PHL194M00, propionibacterium virus PHL199M00, propionibacterium virus PHL301M00, propionibacterium virus PHL308M00, propionibacterium virus Picrate, procrass1, propionibacterium virus SKKY, propionibacterium virus Solid Propionibacterium virus Stormn, propionibacterium virus Wizzo, pseudomonas virus PaMx28, pseudomonas virus PaMx74, mycobacterium virus Patience, mycobacterium virus PBI1, rhodococcus (Rhodococcus) virus Pepy6, rhodococcus virus Poco6, propionibacterium virus PFR1, streptomyces virus phiBT1, streptomyces virus phiC31, streptomyces virus TG1, acinetobacter (Caulobacter) virus Karma, acinetobacter virus Magneto, acinetobacter virus phiCbK, acetobacter virus Rogue, acetobacter virus Swift, staphylococcus virus 11, staphylococcus virus 29, staphylococcus virus 37, staphylococcus virus 53, staphylococcus virus 55, staphylococcus virus 69, staphylococcus virus 71, staphylococcus virus 80, staphylococcus virus 85, staphylococcus virus 88, staphylococcus virus 92, staphylococcus virus 96, staphylococcus virus 187, staphylococcus virus 52a, staphylococcus virus 80alpha, staphylococcus virus CNPH82, staphylococcus virus EW, staphylococcus virus IPLA5, staphylococcus virus IPLA7, staphylococcus virus IPLA88, staphylococcus virus PH15, staphylococcus virus phiETA2, staphylococcus virus phiETA3, staphylococcus virus phiMR11, staphylococcus virus phiMR25, staphylococcus virus phiNM1 staphylococcus virus phiNM2, staphylococcus virus phiNM4, staphylococcus virus SAP26, staphylococcus virus X2, enterococcus virus FL1, enterococcus virus FL2, enterococcus virus FL3, lactobacillus virus ATCC8014, lactobacillus virus phiJL1, pediococcus (Pediococcus) virus cIP1, aeromonas virus pIS A, listeria virus LP302, listeria virus PSA, methanobacter (Methanobacterium) virus psiM1, rhodobacter virus RDJL2, rhodococcus virus RER2, enterococcus virus BC611, enterococcus virus IMEF 1, enterococcus virus SAP6, enterococcus virus VD13, streptococcus virus SPQS1, mycobacterium virus pyrus, mycobacterium virus Send513, burkholderia virus 1, pseudomonas virus 73, kaAb 26, and pseudomonas 25 Khemnas, escherichia virus Cajan, escherichia virus Seurat, staphylococcus virus SEP9, staphylococcus virus Sextaec, streptococcus virus 858, streptococcus virus 2972, streptococcus virus ALQ132, streptococcus virus O1205, streptococcus virus Sfi11, streptococcus virus 7201, streptococcus virus DT1, streptococcus virus phiAbc2, streptococcus virus Sfi19, streptococcus virus Sfi21, paenium virus Diva, paenium virus Hb10C2, paenium virus Rani, paenium virus Shelly, paenium virus Sitara, paenium virus Willow, lactococcus virus 712, lactococcus virus ASCC191, lactococcus virus C273, lactococcus virus C281, lactococcus virus C465, lactococcus virus ASCC532, lactococcus virus BB29, lactococcus virus 38170, bicoccoid virus CB13, bicoccoid 14 lactococcus virus CB19, lactococcus virus CB20, lactococcus virus jj50, lactococcus virus P2, lactococcus virus P008, lactococcus virus sk1, lactococcus virus Sl4, bacillus virus Slash, bacillus virus Stahl, bacillus virus Staley, bacillus virus Stills, gordon virus Bachia, gordon virus ClubL, gordon virus OneUp, gordon virus Smoothia, gordon virus Soups, bacillus virus SPbeta, vibrio virus MAR10, vibrio virus SSP002, escherichia virus AK33, escherichia virus BF23, escherichia virus DT57C, escherichia virus EPS7, escherichia virus FFH1, escherichia virus H8, escherichia virus SLL 09, escherichia virus T5, salmonella virus spp 35, salmonella virus spv 2, and Salmonella virus spp 7, and the like, salmonella virus Stitch, arthrobacter virus Tank, tsukamurella (Tsukamu villa) virus TIN2, tsukamurella virus TIN3, tsukamurella virus TIN4, rhodobacter virus RcSpartan, rhodobacter virus RcTitan, mycobacterium virus Anaya, mycobacterium virus Angelica, mycobacterium virus Crimd, mycobacterium virus Finnbarth, mycobacterium virus Jaws, mycobacterium virus Larva, mycobacterium virus Macncheese, mycobacterium virus Pixie, mycobacterium virus TM4, bacillus virus BMBtp2, bacillus virus TP21, tuber virus Tp84, vitis virus 47, vitis virus 3a, vitis virus 42e, vitis virus LA35, vitis virus phi12, vitis virus Phist, brucella virus 32, mycobacterium virus RGL, mycobacterium virus Hv 3, gata virus Hindle, and Gauss virus Bacillus virus Wbeta, mycobacterium virus Wildcat, gordonia virus Twister6, gordonia virus Wizard, gordonia virus Hotorobo, gordonia virus Monty, gordonia virus Woes, xanthomonas virus CP1, xanthomonas virus OP1, xanthomonas virus phil7, xanthomonas virus Xop, xanthomonas virus Xp10, streptomyces virus TP1604, streptomyces virus YDN12, alpha-Proteobacteria (Alphateobacteria) virus phiJl001, pseudomonas virus LKO4, pseudomonas virus M6, pseudomonas virus MP1412, pseudomonas virus PAE1, pseudomonas virus Yua, pseudomonas virus PM2, pseudomonas virus phi6, pseudomonas virus phi8, pseudomonas phi12, pseudomonas virus phi13, pseudomonas phi29, NN virus phi 54, pseudomonas virus, pseudomonas virus phiYY, vibrio virus fs1, vibrio virus VGJ, ralstonia virus RS603, ralstonia virus RSM1, ralstonia virus RSM3, escherichia virus M13, escherichia virus I22, salmonella virus Ike, acholoproma (Acholoprosma) virus L51, vibrio virus fs2, vibrio virus VFJ, escherichia virus If1, propionibacterium virus B5, pseudomonas virus Pf1, pseudomonas virus Pf3, ralstonia virus PE226, ralstonia virus RSS1, spiroplasma (Spiroplasma) virus SVTS2, sphaeromonas virus PSH1, sphaeromonas virus SMA6, sphaeromonas virus SMA7, sphaeromonas virus SMA9, vibrio virus Xi, vibrio virus KSF1, vibrio virus Vf6, vibrio virus Vf3, vibrio virus 4233, vibrio virus Vf3 Xanthomonas virus Cf1C, helicobacter C74, helicobacter R8A2B, helicobacter SkV CR23x, escherichia FI, escherichia Qbeta, escherichia BZ13, escherichia MS2, escherichia alpha3, escherichia ID21, escherichia ID32, escherichia ID62, escherichia NC28, escherichia NC29, escherichia NC35, escherichia PhiliK, escherichia St1, escherichia WA45, escherichia G4, escherichia ID52, escherichia Talmos, escherichia Philix 174, bdellovibrio MAC1, bdellovibrio 2K, chlamydia Chp, chlamydia 372, AR 34, CPP 1, CPP 23, CPP 1, CPP 2, and CPP 2, achromosoma virus L2, pseudomonas virus PR4, pseudomonas virus PRD1, bacillus virus AP50, bacillus virus Bam35, bacillus virus GIL16, bacillus virus Wip1, escherichia virus phi80, escherichia virus RB42, escherichia virus T2, escherichia virus T3, escherichia virus T6, escherichia virus VT2-Sakai, escherichia virus VT1-Sakai, escherichia virus CP-933V, escherichia virus P27, escherichia virus Stx2phi-I, escherichia virus Stx1phi the Escherichia virus Stx2phi-II, the Escherichia virus CP-1639, the Escherichia virus based BP-4795, the Escherichia virus 86, the Escherichia virus Min27, the Escherichia virus 2851, the Escherichia virus 1717, the Escherichia virus YYZ-2008, the Escherichia virus EC026_P06, the Escherichia virus ECO103_P15, the Escherichia virus ECO103_P12, the Escherichia virus ECO111_P16, the Escherichia virus ECO111_P11, the Escherichia virus VT2phi_272, the Escherichia virus TL-2011c, the Escherichia virus P13374, and the Escherichia virus Sp5; the first bacteriophage is different from the second bacteriophage.

In one embodiment, the first bacteriophage is selected from the group consisting of: BW73、B278、D6、D108、E、El、E24、E41、FI-2、FI-4、FI-5、HI8A、Ffl8B、i、MM、Mu、025、PhI-5、Pk、PSP3、Pl、PlD、P2、P4、Sl、 7A、/>18. 28-1, 186, 299, H-Escherichia (2), AB48, CM, C4, C16, DD-VI, E4, E7, E28, FIl, FI3, H, hl, H3, H8, K3, M, N, ND-2, ND-3, ND4, ND-5, ND6, ND-7, ox-I, ox-2, ox-3, ox-4, ox-5, ox-6, phI-I, RB, RB43, RB49, RB69, S, saI-I, sal-2, sal-3, sal-4, sal-5, sal-6, TC23, TC45, tuII-6, tuIP-24, tuII-46, tuIP-60, T2, T4, T6, T35, αl, 1, IA, 3A, 3T+,>9266Q、CFO103、HK620、J、K、KlF、m59、no.A、no.E、no.3、no.9、N4、sd、T3、T7、WPK、W31、ΔH、/> Φ04-CF、Φ05、Φ06、Φ07、/> omega 8, 1, 3, 7, 8, 26, 27, 28-2, 29, 30, 31, 32, 38, 39, 42, 933W, NN-Escherichia (1), esc-7-11, AC30, CVX-5, cl, DDUP, ECl, EC, E21, E29, fl, F26S, F S, hi, HK022, HK97, HK139, HK253, HK256, K7, ND-I, PA-2, q, S2, tl,), CVX-5, cl, DDUP, ECl, EC, E21, E29, fl, F26S, F S, hi, HK022, HK97, HK139, K253, HK256, K7, ND-I, PA-2, q, S2, tl,)T3C、T5、UC-I、w、β4、γ2、λ、ΦD326、/>Φ06、Φ7、Φ10、/>χ, 2, 4A, 6, 8A, 102, 150, 168, 174, 3000, AC6, AC7, AC28, AC43, AC50, AC57, AC81, AC95, HK243, klO, ZG/3A, 5A, 21EL, H19-J, and 933H.

In a preferred embodiment, the lytic bacteriophage is a T7 bacteriophage. In another preferred embodiment, the lytic bacteriophage is a dermatophyte acnes lytic bacteriophage.

In a preferred embodiment, the non-lytic bacteriophage is a lambda bacteriophage. In another preferred embodiment, the non-lytic bacteriophage is a propionibacterium freudenreichii bacteriophage.

In a preferred embodiment, the lytic bacteriophage is a T7 bacteriophage and the non-lytic bacteriophage is a lambda bacteriophage. In another preferred embodiment, the lytic bacteriophage is a dermatophyte acne lytic bacteriophage and the non-lytic bacteriophage is a propionibacterium freudenreichii bacteriophage.

Additional bacterial genes

As is well known to the skilled person, some phages use products produced by their bacterial hosts to fold and/or assemble their structural elements and/or for proper packaging of their DNA.

Thus, in a particular embodiment, the production bacterial cell further comprises at least one bacterial gene derived from a bacterial species or strain from which the lytic bacteriophage originates, involved in folding and/or assembly of the bacteriophage structural elements and/or involved in DNA packaging.

As the skilled artisan will appreciate, the bacterial genes involved in the folding and/or assembly of the phage structural elements depend on the particular bacteriophage from which the phage structural elements are obtained. They generally include bacterial genes encoding chaperones.

Similarly, the bacterial genes involved in phage DNA packaging depend on the particular bacteriophage from which the phage DNA packaging genes are obtained. Examples of such bacterial genes include genes encoding IHF proteins.

Payload

In particular embodiments, the producer bacterial cell further comprises a payload to be packaged into the phage particle or phage-derived delivery vehicle.

As used herein, the term "payload" refers to any nucleic acid sequence (DNA and/or RNA) or amino acid sequence or combination of both (such as, without limitation, peptide nucleic acid or peptide-oligonucleotide conjugate) that is transferred into bacteria with a delivery vehicle. In a particular embodiment, the payload is a nucleic acid payload, more particularly a DNA and/or RNA payload, still particularly a DNA payload.

The term "payload" may also refer to a plasmid, vector or cargo.

The payload may be a phagemid or plasmid obtained from the natural, evolved or engineered phage genome. The payload may also consist only in part of phagemids or plasmids obtained from the natural, evolved or engineered phage genome.

As used herein, the term "phagemid" or "plasmid" is equivalent and refers to a recombinant DNA vector comprising at least one bacteriophage genome sequence and is capable of allowing packaging in a capsid, and which preferably is incapable of producing offspring, more particularly a vector derived from a plasmid and bacteriophage genome. The phagemid of the present disclosure comprises a phage packaging site and optionally an origin of replication (ori), in particular an origin of replication of a bacterium and/or phage. In one embodiment, the phagemid does not comprise an origin of replication and is therefore unable to replicate itself once injected into bacteria. Alternatively, the phagemid comprises a plasmid replication origin, in particular a bacterial and/or phage replication origin.

In certain embodiments, the payload will be packaged in the form of a packaged phagemid.

As used herein, the term "packaged phagemid" refers to a phagemid encapsulated in a bacteriophage scaffold, phage-derived delivery particle or capsid. In particular, it refers to a bacteriophage scaffold, bacteriophage delivery particle or capsid that is devoid of a bacteriophage genome. Packaged phagemids can be generated using helper phage strategies well known to those skilled in the art. Helper phages generally comprise all genes encoding structural and functional proteins, which are essential for the phagemid of the invention to be packaged.

In certain embodiments, the payload is to be delivered into a targeted bacterial cell as defined below.

In more particular embodiments, the payload is stably maintained in the targeted bacterial cell. In alternative embodiments, the payload is not replicated in the targeted bacterial cell.

Sequences of interest under the control of a promoter

In certain embodiments, the payload comprises a sequence of interest, particularly under the control of a promoter.

Promoters may be classified as strong or weak according to their affinity for RNA polymerase, as known to those skilled in the art. The strength of a promoter may depend on whether transcription initiation occurs at the promoter at a high or low frequency. Different promoters with different intensities may be used in the present invention, resulting in different levels of gene/protein expression (e.g., lower levels of expression starting from mRNA derived from a weak promoter than from a strong promoter).

One of ordinary skill in the art will appreciate that the promoter sequence may be selected from a number of known bacterial genes expressed by various bacterial species. Furthermore, methods of prokaryotic promoter prediction exist and can be based on DNA stability analysis, as described by Kanhere and Bansal (BMC Bioinformatics 2005, 6:1). Thus, selection of promoters on payloads used in the context of the present invention may be made based on the bacteria targeted.

In some embodiments, the nucleic acid of interest may be under the control of a recombinant or heterologous promoter, which refers to a promoter that is not normally associated with the nucleic acid of interest in its natural environment.

Examples of bacterial promoters for use in accordance with the invention include, without limitation, positive regulated E.coli promoters such as the positive regulated sigma 70 promoter (e.g., inducible pBad/araC promoter, lux cassette right promoter, modified lambda Prm promoter, plac Or2-62 (positive), pBad/AraC, pBad, P (Las) TetO with additional REN sites, P (Las) CIO, P (Rhl), pu, fecA, pRE, cadC, hns, pLas, pLux), "s" promoter (e.g., pdps), sigma 32 promoter (e.g., heat shock) and sigma 54 promoter (e.g., glnAP 2); a negative regulated E.coli promoter, such as negatively regulated sigma 70 promoters (e.g.promoter (PRM+), modified lambda Prm promoter, tetR-TetR-4C P (Las) TetO, P (Las) CIO, P (Lac) IQ, recA_DlexO_DLac01, dapAp, fecA, pspac-hy, pel, pLux-cl, plus-Lac, cinR, cinL, glucose control, modified Pr, modified Prm+, fecA, pcya, recA (SOS), recA (SOS), emrR_regulated, betl_regulated, pLac_lux, emrR_Lac pLac/Mnt, pTet/Mnt, lsrA/cI, pLux/cI, lacl, lacIQ, pLacIQl, pLas/cI, pLas/Lux, pLux/Las, pRECA with LexA binding site, reverse BBa_R0011, pLacI/ara-1, pLacIq, rrnB PI, cadC, hns, pfhuA, pBad/araC, nhaA, ompF, rcnR), sigma S promoter (e.g. Lutz-Bujar LacO with selective sigma factor sigma 38), sigma 32 promoter (e.g. Lutz-Bujar LacO with selective sigma factor sigma 32), sigma 54 promoter (e.g. glnAP 2); negative regulated bacillus subtilis promoters such as the repressed bacillus subtilis sigma a promoter (e.g., gram positive IPTG inducible, xyl, hyper-spray), sigma promoter and the BioFAB promoter disclosed in Mutalik VK et al (Nature Methods,2013,10:354-360, see, inter alia, supplementary data) and on the BioFAB website (http:// bioofab. Synbec. Org/data). Other inducible microbial promoters and/or bacterial promoters may be used in accordance with the present invention. Inducible promoters for use in accordance with the present disclosure may be induced (or repressed) by one or more physiological conditions, such as pH, temperature, radiation, osmotic pressure, saline gradients, cell surface binding, and changes in the concentration of one or more exogenous or endogenous inducers. The exogenous inducer or inducer can include, without limitation, amino acids and amino acid analogs, sugars and polysaccharides, nucleic acids, protein transcription activators and repressors, cytokines, toxins, petroleum-based compounds, metal-containing compounds, salts, ions, enzyme substrate analogs, hormones, or combinations thereof.

Particularly preferred bacterial Promoters for use according to the invention may be selected from constitutive Promoters regulated by sigma 70, such as the Promoters of the Anderson collection (http:// parts. Igem. Org/Promoters/catalyst/Anderson): BBa J23100, BBa J23101, BBa J23102, BBa J23103, BBa J23104, BBa J23105, BBa J23106, BBa J23107, BBa J23108, BBa J23109, BBa J23110, BBa J23111, BBa J23112, BBa J23113, BBa J23114, BBa J23115, BBa J23116, BBa J23117, BBa J23118, and BBa J23119.

Other preferred bacterial promoters are those disclosed in Stanton et al, (2014) Nat.chem.biol.10:99-105 (incorporated herein by reference), including in particular TetR, icaR (A), amtR, betI, srpR, orf2, BM3R1, butR, phlF, psrA, hlyIIR, ameR, lmrA, qacR, scbR, mcbR, litR, hapR, smcR, tarA and variants thereof. In particular embodiments, the promoter is SrpR and/or PhlF or variants thereof.

In some embodiments of the invention, a promoter may or may not be used with an "enhancer," which refers to a cis-acting regulatory sequence involved in transcriptional activation of a nucleic acid sequence downstream of the promoter. Enhancers may be located at any functional position before or after the promoter.

In some embodiments, the payload may comprise a terminator sequence or terminator. As used herein, a "terminator" is a nucleic acid sequence that causes transcription to stop. The terminator may be unidirectional or bidirectional. It consists of a DNA sequence involved in the specific termination of RNA transcripts by RNA polymerase. The terminator sequence prevents transcriptional activation of the downstream nucleic acid sequence by the upstream promoter. Thus, in certain embodiments, a terminator is contemplated that ends the production of the RNA transcript. Terminators may be necessary to achieve desired gene/protein expression levels in vivo.

The most common type of terminator is the forward terminator. When placed downstream of a normally transcribed nucleic acid of interest, a forward transcription terminator will cause transcription to cease. In some embodiments, a bi-directional transcription terminator is provided that generally causes transcription termination on both the forward and reverse strands. In some embodiments, a reverse transcription terminator is provided, which generally terminates transcription only on the reverse strand. In prokaryotic systems, terminators generally fall into two categories, (1) ρ -independent terminators and (2) ρ -dependent terminators. The ρ -independent terminator usually consists of a stem-loop forming palindromic sequence enriched with G-C base pairs followed by a string of uracil bases.

The terminator for use according to the present invention includes any transcription terminator described herein or known to one of ordinary skill in the art. Examples of terminators include, without limitation, termination sequences of genes such as bovine growth hormone terminators and viral termination sequences such as TO terminators, TE terminators, λTl and T1T2 terminators found in bacterial systems. In some embodiments, the termination signal may be a sequence that cannot be transcribed or translated, such as those resulting from sequence truncation.

The terminator for use according to the present invention also includes the terminator disclosed in Chen YJ et al (2013,Nature Methods,10:659-664) and the BioFAB terminator disclosed in Camfray G et al (Nucl Acids Res,2013,41 (9): 5139-5148).

In one embodiment, the sequence of interest is a programmable nuclease circuit to be delivered to the targeted bacteria. Such a programmable nuclease loop may be capable of mediating sequence-specific elimination of bacteria containing a target gene of interest (e.g., a gene that is detrimental to humans). Some embodiments of the present disclosure relate to engineered variants of the type II CRISPR-Cas (clustered regularly interspaced short palindromic repeat-CRISPR-associated) system of streptococcus pyogenes (Streptococcus pyogenes). Other programmable nucleases that can be used include other CRISPR-Cas systems, engineered TALEN (transcription activator-like effector nuclease) variants, engineered Zinc Finger Nuclease (ZFN) variants, natural, evolved or engineered meganuclease or recombinase variants, and any combination or hybrid of programmable nucleases. Thus, the autonomously designed and engineered distributed loop provided herein can be used to selectively excise DNA encoding a gene of interest, such as a toxin gene, virulence factor gene, antibiotic resistance gene, remodeling gene or regulatory gene (see WO2014124226 and US 2015/0064138).

Other sequences of interest (preferably programmable) may be added to the payload in order to be delivered to the targeted bacteria. Preferably, the sequence of interest added to the payload results in cell death of the targeted bacteria. For example, a nucleic acid sequence of interest added to a payload may encode perforins, endolysins, restriction enzymes, or toxins that affect the targeted bacteria.

Alternatively, the sequence of interest added to the payload does not result in death of the targeted bacteria. For example, the sequence of interest may encode a reporter gene that results in a luminescent or fluorescent signal. Alternatively, the sequences of interest may comprise proteins and enzymes that fulfill useful functions, such as altering the metabolism of the targeted bacteria, the composition of its environment, or affecting the host subject. More specifically, the sequence of interest may be an antigen that triggers an immune response in the host immune subject. The specific antigen may be released in the environment after induction of lysis of the target cells, or may be secreted by the target cells (Costa et al, nat Rev Microbiol.2015Jun;13 (6): 343-59; ann et al Curr Top Microbiol Immunol.2017; 404:267-308).

In particular embodiments, the nucleic acid sequence of interest is selected from Cas nuclease, cas9 nuclease, guide RNA, single guide RNA (sgRNA), CRISPR locus, genes expressing enzymes such as nucleases or kinases, TALENs, ZFNs, meganucleases, recombinases, transposases, bacterial receptors, membrane proteins, structural proteins, secreted proteins, genes expressing resistance to antibiotics or general drugs, genes expressing toxic proteins or toxic elements, and genes expressing virulence proteins or virulence elements, bacterial secreted proteins or transporters, bacterial pores, or any combination thereof. These proteins may also be modified or engineered to include additional features such as the addition or removal of a function (e.g., dCas 9), addition of secretion signals to proteins that are not normally secreted, addition of exogenous peptides in the loop, as non-limiting examples.

In particular embodiments, the nucleic acid sequence of interest encodes a guide RNA assisted targeting (INTEGRATE) system, generally as in Vo et al, nat biotechnol 2021apr;39 (4) 480-489, the INTEGRATE system includes, for example, a Vibrio cholerae type I (V.cholerae) CRISPR transposon or a V-K pseudoclade greetings (Scytonema hofmanii) CRISPR transposon. In particular embodiments, the nucleic acid sequence of interest includes nucleic acids encoding crrnas, nucleic acids encoding TniQ cascade, cas8, cas7 and cas6 proteins, nucleic acids encoding tnsA, tnsB and tnsC proteins, and further includes donor DNA encoding a protein of interest to be added to the targeted bacterial genome. In particular embodiments, the nucleic acids encoding the TniQ cascade, cas8, cas7 and cas6 proteins and encoding the tnsA, tnsB and tnsC proteins are in the form of a single polycistronic nucleic acid. In another particular embodiment, the nucleic acid sequence of interest includes a nucleic acid encoding a guide RNA, a nucleic acid encoding cas12k protein, tnsB and tnsC proteins, and a TniQ cascade, and further includes donor DNA encoding a protein of interest to be added to the targeted bacterial genome.

In particular embodiments, the payloads used in the context of the present invention comprise a sequence of interest encoding a bacteriocin, which may be a protein toxin produced by a bacterium to kill or inhibit the growth of other bacteria. Bacteriocins are classified in several ways, including producer strains, common resistance mechanisms, and killing mechanisms. Such bacteriocins from gram-negative bacteria (e.g., colicins, and tail bacteriocins (tailocins)) and from gram-positive bacteria (e.g., class I, II, III, or IV bacteriocins) have been described.

In one embodiment, the payload used in the context of the present invention further comprises a sequence of interest encoding a toxin selected from the group consisting of: colicin-like bacteriocins, cerocins, class I, class II, class III and class IV bacteriocins. The circuit may also encode transport proteins required to secrete toxins into the extracellular space.

In particular embodiments, the corresponding immune polypeptides (i.e., antitoxins) can be used to protect bacterial cells (see a review of Cotter et al, nature Reviews Microbiology 11:95, 2013) for payload generation and encapsidation purposes, but the payloads used in the context of pharmaceutical compositions and the present invention are not present in the targeted bacteria in which they are delivered.

In particular embodiments, the payload used in the context of the present invention comprises a sequence of interest encoding a CRISPR-Cas system.

The CRISPR-Cas system contains two different elements, i.e., i) an endonuclease, in this case a CRISPR-associated nuclease (Cas or "CRISPR-associated protein"), and ii) a guide RNA. Depending on the type of CRISPR system, the guide RNA may be in the form of a chimeric RNA consisting of a combination of CRISPR (crRNA) bacterial RNA and tracrRNA (transactivation RNA CRISPR) (Jinek et al, science.2012Aug17;337 (6096): 816-21). The guide RNA binds to the targeting specificity of the crRNA corresponding to the "spacer" that serves as a guide for the Cas protein, as well as the conformational nature of the tracrRNA in a single transcript. When the guide RNA is expressed simultaneously with the Cas protein in the cell, the target genomic sequence may be permanently interrupted (and, depending on the location, cause disappearance of the targeting and surrounding sequences and/or cell death) or modified. Modification may be guided by the repair matrix.

Depending on the mechanism of action of nucleases, CRISPR-Cas systems include two main categories:

class-1 consists of multi-subunit effector complexes and includes I, III and type IV;

class-2 consists of single subunit effector modules such as Cas9 nucleases and includes type II (II-A, II-B, II-C, II-C variants), type V (V-A, V-B, V-C, V-D, V-E, V-U1, V-U2, V-U3, V-U4, V-U5) and type VI (VI-A, VI-B1, VI-B2, VI-C, VI-D).

The sequence of interest according to the invention may comprise a nucleic acid sequence encoding a Cas protein. A variety of CRISPR enzymes are useful as sequences of interest on payloads used in the context of the present invention. In some embodiments, the CRISPR enzyme is a type II CRISPR enzyme, a type II-a or a type II-B CRISPR enzyme. In another embodiment, the CRISPR enzyme is a type I CRISPR enzyme or a type III CRISPR enzyme. In some embodiments, the CRISPR enzyme catalyzes DNA excision. In some other embodiments, the CRISPR enzyme catalyzes RNA excision. In one embodiment, the CRISPR enzyme can be coupled to a guide RNA or a single guide RNA (sgRNA). In certain embodiments, the guide RNA or sgRNA targets a gene selected from the group consisting of: antibiotic resistance genes, virulence protein or factor genes, toxin protein or factor genes, bacterial receptor genes, membrane protein genes, structural protein genes, secreted protein genes, genes that express resistance to general drugs, and genes that cause deleterious effects on the host subject.

The sequence of interest may comprise a nucleic acid sequence encoding a guide RNA or sgRNA to guide Cas protein endogenous to the targeted bacteria, alone or in combination with a Cas protein and/or guide RNA encoded by the payload.

Non-limiting examples of Cas proteins that are part of a multi-subunit effector or as a single subunit effector include Cas1, cas1B, cas2, cas3, cas4, cas5, cas6, cas7, cas8, cas9 (also known as Csn1 and Csx 12), cas10, cas11 (SS), cas12a (Cpf 1), cas12b (C2C 1), cas12C (C2C 3), cas12d (CasY), cas12e (CasX), C2C4, C2C8, C2C5, C2C10, C2C9, cas13a (C2), cas13b (C2C 6), cas13C (C2C 7), cas13d, csa5, csc 1' Csc2, cse1, cse2, csy3, csf1, csf2, csf3, csf4, csm1, csm2, csm3, csm4, csm5, csm6, cmr1, cmr3, cmr4, cmr5, cmr6, csn2, csb1, csb2, csb3, csx17, csx14, csx10, csx16, csax, csx13, csx1, csx15, sdcpf1, cttCpf 1, tspCpf 1, cmaCpf1, pcpf 1, erCpf1, fbcCpf 1, asCpf1, lbCpf1, mad4, mad7, cms1, homologs thereof, orthologs thereof, variants thereof, or modified versions thereof. In some embodiments, the CRISPR enzyme cleaves both strands of the target nucleic acid at a pre-spacer adjacent to a motif (PAM) site.

In particular embodiments, the CRISPR enzyme is any Cas9 protein, such as any naturally occurring bacterial Cas9 and any variant, homolog or ortholog thereof.

By "Cas9" is meant a protein Cas9 (also known as Csn1 or Csx 12) or a functional protein, peptide or polypeptide fragment thereof, i.e. capable of interacting with the guide RNA(s) and exerting an enzymatic activity (nuclease) allowing it to double-strand cleave the DNA of the target genome. "Cas9" may thus represent a modified protein, e.g. a protein domain that is truncated to remove a domain not essential for the intended function of the protein, in particular a domain not essential for interaction with the gRNA(s).

As used in the context of the present invention, sequences encoding Cas9 (whole protein or fragments thereof) can be obtained from any known Cas9 protein (Fonfara et al, nucleic Acids Res.2014Feb;42 (4): 2577-90; koonin et al, currOpinMicrobiol.2017Jun; 37:67-78). Examples of Cas9 proteins useful in the present invention include, but are not limited to, cas9 proteins of: streptococcus pyogenes (SpCas 9), streptococcus thermophilus (Streptococcus thermophiles) (St 1Cas9, st3Cas 9), streptococcus mutans (Streptococcus mutans), staphylococcus aureus (Staphylococcus aureus) (SaCas 9), campylobacter jejuni (Campylobacter jejuni) (CjCas 9), francissamum newlare (Francisella novicida) (FnCas 9), and neisseria meningitidis (Neisseria meningitides) (NmCas 9).

As used in the context of the present invention, the sequence encoding Cpfl (Cas 12 a) (whole protein or fragment thereof) may be obtained from any known Cpfl (Cas 12 a) protein (Koonin et al, currOpinMicrobiol.2017Jun; 37:67-78). Examples of Cpf1 (Cas 12 a) proteins useful in the present invention include, but are not limited to, the following Cpf1 (Cas 12 a) proteins: amino acid coccus species (Acidaminococcus sp), bacteria of the family chaetoceraceae (Lachnospiraceae bacteriu) and new inland francisco.

As used in the context of the present invention, sequences encoding Cas13a (whole protein or fragments thereof) can be obtained from any known Cas13a (C2C 2) protein (Abudayyeh et al, nature.2017Oct 12;550 (7675): 280-284). Examples of Cas13a (C2) proteins useful in the present invention include, but are not limited to, the following Cas13a (C2) proteins: wei De ciliated (Leptotrichia wadei) (LwaCas 13 a).

As used in the context of the present invention, sequences encoding Cas13d (whole protein or fragments thereof) can be obtained from any known Cas13d protein (Yan et al Mol cell.2018Apr 19;70 (2): 327-339.e5.). Examples of Cas13d proteins useful in the present invention include, but are not limited to, cas13d proteins of: eubacterium inertens (Eubacterium siraeum) and Ruminococcus species (Ruminococcus sp.).

As used in the context of the present invention, the sequence encoding Mad4 (whole protein or fragment thereof) is disclosed in international application WO 2018/236548.

As used in the context of the present invention, the sequence encoding Mad7 (whole protein or fragment thereof) is disclosed in international application WO 2018/236548.

As used in the context of the present invention, sequences encoding Cms1 (whole protein or fragments thereof) are disclosed in international patent application WO 2017/141173.

In particular embodiments, the nucleic acid sequence of interest is a CRISPR/Cas9 system for reducing expression or inactivating a gene selected from the group consisting of: antibiotic resistance genes, virulence factors or protein genes, toxin factors or protein genes, genes expressing bacterial receptors, membrane proteins, structural proteins, secreted proteins, genes expressing resistance to general drugs, and genes causing deleterious effects on the host subject.

In one embodiment, the CRISPR system is used to target and inactivate virulence factors. Virulence factors can be any substance produced by a pathogen that alters host subject-pathogen interactions by increasing the extent of damage made to the host subject. Virulence factors are used by pathogens in a number of ways including, for example, in the adhesion or colonization of cells in a niche (niche) in a host subject, to evade the host subject's immune response, to promote entry into and exit from cells in the host subject, to gain nutrition from the host subject, or to inhibit other physiological processes in the host subject. Virulence factors can include enzymes, endotoxins, adhesion factors, motility factors, factors involved in complement evasion, clearance factors, and factors that promote biofilm formation. For example, such targeted virulence factor genes may be escherichia coli virulence factor genes, such as, without limitation, EHEC-HlyA, stx1 (VT 1), stx2 (VT 2), stx2a (VT 2 a), stx2b (VT 2 b), stx2c (VT 2 c), stx2d (VT 2 d), stx2e (VT 2 e) and Stx2F (VT 2F), stx2h (VT 2 h), stx2k, fimA, fimF, fimH, neuC, kpsE, sfa, foc, iroN, aer, iha, papC, papGI, papGII, papGIII, hlyC, cnf1, hra, sat, ireA, usp ompT, ibeA, malX, fyuA, irp2, traT, afaD, ipaH, eltB, estA, bfpA, eaeA, espA, aaiC, aatA, TEM, CTX, SHV, csgA, csgB, csgC, csgD, csgE, csgF, csgG, csgH, T1SS, T2SS, T3SS, T4SS, T5SS, T6SS (secretion system). For example, such targeted virulence factor genes can be shigella dysenteriae (Shigella dysenteriae) virulence factor genes, such as, without limitation, stx1 and stx2. For example, such targeted virulence factor genes can be Yersinia pestis (Yersinia pestis) virulence factor genes, such as, without limitation, yscF (plasmid-borne (pCDl) T3SS outer needle subunit (external needle subunit)). For example, such a targeted virulence factor gene may be a Francisella tularensis (Francisella tularensis) virulence factor gene, such as, without limitation, fslA. For example, such targeted virulence factor genes can be bacillus anthracis (Bacillus anthracis) virulence factor genes, such as, without limitation, pag (anthrax toxin, cell-binding protective antigen). For example, such targeted virulence factor genes can be Vibrio cholerae (Vibrio cholera) virulence factor genes, such as, without limitation, ctxA and ctxB (cholera toxin), tcpA (toxin co-regulated pili), and toxT (major virulence regulator). For example, such targeted virulence factor genes may be pseudomonas aeruginosa (Pseudomonas aerigunosa) virulence factor genes, such as, without limitation, pyovine (e.g., sigma factor pvdS, biosynthesis gene pvdL, pvdl, pvdJ, pvdH, pvdA, pvdF, pvdQ, pvdN, pvdM, pvdO, pvdP, transporter gene pvdE, pvdR, pvdT, opmQ), siderophores pyochelin (e.g., pchD, pchC, pchB, pchA, pchE, pchF and pchG), and toxins (e.g., exoU, exoS, and exoT). For example, such targeted virulence factor genes can be klebsiella pneumoniae (Klebsiella pneumoniae) virulence factor genes, such as, without limitation, fimA (adhesion, type I bacteria Mao Zhuyao subunit) and cps (capsular polysaccharide). For example, such targeted virulence factor genes can be acinetobacter baumannii (Acinetobacter baumannii) virulence factor genes, such as, without limitation, ptk (capsular polymerization) and epsA (assembly). For example, such targeted virulence factor genes can be salmonella enterica typhi (Salmonella enterica Typhi) virulence factor genes, such as, without limitation, MIA (invasion, SPI-1 regulator), ssrB (SPI-2 regulator), and those associated with bile tolerance, including the efflux pump genes acrA, acrB, and tolC. For example, such targeted virulence factor genes can be fusobacterium nucleatum (Fusobacterium nucleatum) virulence factor genes, such as, without limitation, fadA and TIGIT. For example, such targeted virulence factor genes can be bacteroides fragilis (Bacteroides fragilis) virulence factor genes, such as, without limitation, bft. For example, such targeted virulence factor genes may be the dermatobacterium acnes (Cutibacterium acnes) porphyrin gene, the CAMP factors (CAMP 1, CAMP2, CAMP3, CAMP 4), the hyaluronate lyase (hyt-IB/II, hyt-IA), the lipase (GehA, gehB), the lysin, the sialidases, the gangliosidase, the endo- β -N-acetylglucosaminidase, the dermatan sulfate adhesins (DsA 1, dsA 2), the proline-threonine repeat (PTR) or any virulence factor included on the acne-related genomic loci 1, 2, 3 (plasmids), 4 (such as the compact adhesion locus (tad)), the streptolysin S-related gene (sag), the non-ribosomal peptide synthase (NRPS), such as Tomida et al, mhio.2013 apr 30;4 (3) e 00003-13.

In another embodiment, CRISPR/Cas systems are used to target and inactivate antibiotic resistance genes such as, without limitation, gyrB, parE, parY, AAC (1), AAC (2 '), AAC (3), AAC (6'), ANT (2 "), ANT (3"), ANT (4 '), ANT (6), ANT (9), APH (2 "), APH (3'), APH (4), APH (6), APH (7), APH (9), armA, rmtA, rmtB, rmtC, sgm, AER, BLA1, CTX-M, KPC, SHV, TEM, blaB, ccrA, IMP, NDM, VIM, ACT, ampC, CMY, LAT, PDC, OXA β -lactamase, mecA, omp36, ompF, PIB, bla (blaI, blaR 1) and mec (mecI, mecR 1) operons, chloramphenicol Acetyl Transferase (CAT), chloramphenicol phosphotransferase, ethylamine resistant arabinosyl transferase (EmbB), mupA, mupB, integrin (2), cfr rrmethyl transferase, rifampin-ADP-transferase (4), arpr-transferase, monokinetin, ADP-transferase, dfor, dfirp-2), znafalabas, zna, vgb, 75, noroxymatriptase, zna, 75, noroxymatriptase, virginase, 37, stroma, quinazoline (75, and the like FomA, fomB, fosC, fosA, fosB, fosX, vanA, vanB, vanD, vanR, vanS, lincoamide nucleotide transferase (Lin), ereA, ereB, gimA, mgt, ole, macrolide Phosphotransferase (MPH), mefA, mefE, mel, streptothricin acetyl transferase (sat), sul1, sul2, sul3, sulfanilamide resistance FolP, tetracycline inactivating enzyme TetX, tetA, tetB, tetC, tet30, tet31, tetM, tetO, tetQ, tet32, tet36, macAB-TolC, msbA, msrA, vgaB, emrD, emrAB-TolC, norB, gepA, mepA, adeABC, acrD, mexAB-OprM, mtrCDE, emrE, adeR, acrR, baeSR, mexR, phoPQ, mtrR, or any antibiotic resistance gene described in the comprehensive antibiotic resistance database (CARD https:// card.mcmaster.ca /).

In another embodiment, the CRISPR/Cas system is used to target and inactivate bacterial toxin genes. Bacterial toxins may be classified as either exotoxins or endotoxins. Exotoxins are produced and actively secreted; endotoxin remains part of the bacteria. The response to bacterial toxins may involve severe inflammation and may lead to sepsis. Such toxins may be, for example, botulinum neurotoxin, tetanus toxin, staphylococcal toxin, diphtheria toxin, anthrax toxin, alpha toxin, pertussis toxin, shiga toxin, thermostable enterotoxin (e.g., ST), colicin (colibacillus), BFT (bacteroides fragilis toxin), or any of the toxins described in Henkel et al (Toxins from Bacteria in exs.2010; 100:1-29).

In particular embodiments, the payload used in the context of the present invention comprises a sequence of interest encoding a base editing system.

Base Editing (BE) refers to the ability to substitute another base pair for a particular nucleotide base pair on a DNA or RNA molecule. Until recently, the only way to specifically substitute DNA in vivo was to use recombination of template DNA with the locus of interest that carries specific base pair changes. Base editing techniques rely on disparate strategies. Instead of DNA exchange, an enzymatic reaction converts a nucleotide to another nucleotide, resulting in a mismatch at the dsDNA level, which is then corrected by the cellular machinery.

In some embodiments, the base editing system includes one or more of the following enzymes and systems:

a) Cytosine Base Editors (CBEs) and Adenosine Base Editors (ABEs), as described by Rees, H.A.& Liu, d.r.nat Rev Genet 19,770-788 (2018).

To date, seven types of DNA base editors have been described:

cytosine Base Editor (CBE) for converting C:G to T:A (Komor, A et al Nature 533:420-4 (2016))

Adenine base editor (Gaudelli, N.M. et al, nature 551 (7681) 464-471 (2017))

Cytosine Guanine Base Editor (CGBE) for converting C: G to C (Chen, L et al Precise and programmable C: G to G: C base editing in genomic DNA. Biorxiv (2020.; kurt, I et al, CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells. Nature Biotechnology (2020))

Cytosine guanine base editor (Zhao, D et al New base editors change C to A in bacteria and C to G in mammalian cells Nature Biotechnology (2020)) for converting C: G to A: T

Adenine cytosine base editor converting A: T to C: G (WO 2020181180)

Adenine thymine base editor for converting A: T to T: A (WO 2020181202)

Thymine Adenine Base Editor (TABE) for converting T: A to A: T (WO 2020181193; WO2020181178; WO 2020181195)

Base editors differ in terms of base modifying enzymes. CBE relies on ssDNA cytidine deaminase, wherein: APOBEC1, rAPOBEC1, mutants or evolved versions of APOBEC1 (evoAPOBEC 1), APOBEC homologs (APOBEC 3A (eA 3A), anc 689), cytidine deaminase 1 (CDA 1), evoCDA1, FERNY, evoFERNY.

ABE relies on deoxyadenosine deaminase activity fused in tandem to TadA-TadA, an evolved version of TadA, which is an escherichia coli tRNA adenosine deaminase capable of converting adenosine to inosine on ssDNA. TadA includes TadA-8a-e and TadA-7.10.

In addition to base modifying enzymes, there are modifications made to the base editor to improve editing efficiency, accuracy and modularity:

-adding one or two uracil DNA glycosylase inhibitor domains (UGIs) to prevent base excision repair mechanisms from restoring base editing

Addition of Mu-GAM, which reduces the insertion-deletion rate by inhibiting non-homologous end joining mechanisms (NHEJ) in cells

Cas9 with nickase activity (nCas 9D 10A) is used, which facilitates its repair by making a nick on the non-editing strand, thus fixing the edited base.

Use of different Cas proteins from e.g. different organisms, mutants with different PAM motifs or different fidelity or different families (e.g. Cas12 a).

Non-limiting examples of DNA-based editor proteins include BE1, BE2, BE3, BE4-GAM, HF-BE3, marksman-BE 3, target-AID-NG, ABE, EE-BE3, YE1-BE3, YE2-BE3, YEE-BE3, BE-PLUS, saBE3, saBE4-GAM, sa (KKH) -BE3, VQR-BE3, VRER-BE3, EQR-BE3, xBE3, cas12a-BE, ea3A-BE 3A 3A-BE3, TAM, CRISPR-X, ABE7.9, ABE7.10, xABE, ABESa, VQR-ABE, VRER-ABE, sa (KKH) -ABE, ABE8e, spRY-ABE, spRY-CBE, spG-CBE4, spG-ABE, spRY-CBE4, spCas9-NG-ABE, spCas9-NG-CBE4, enasbe1.1, enasbe1.2, enasbe1.3, enasbe1.4, asbes1.1, asbest 1.4, CRISPR-Abest, CRISPR-Cbest, eA3A-BE3, ancbE4.

The Cytosine Guanine Base Editor (CGBE) consists of a nicking enzyme CRISPR fused to:

cytosine deaminase (rAPOBEC) and base excision repair proteins (e.g. rXRCC 1) (Chen, L et al Precise and programmable C: G to G: C base editing in genomic DNA. Biorxiv (2020); chen et al Nature Communications12:1384 (2021))

Rat APOBEC1 variant (R33A) protein and uracil DNA N-glycosylase (eUNG) from E.coli (Kurt, I et al, CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells. Nature Biotechnology (2020))

The Cytosine Adenine Base Editor (CABE) consists of Cas9 nickase, cytidine deaminase (e.g., AID) and uracil-DNA glycosylase (Ung) (Zhao, D et al New base editors change C to A in bacteria and C to G in mammalian cells.

ACBE comprises a nucleic acid programmable DNA binding protein and adenine oxidase (WO 2020181180).

ATBE consists of a Cas9 nickase and one or more adenosine deaminase or oxidase domains (WO 2020181202).

Tab consists of Cas9 nickase and an adenosine methyltransferase, thymine transferase or adenosine deaminase domain (WO 2020181193; WO2020181178; WO 2020181195).

The base editor molecule may also consist of two or more of the above listed editor enzymes (e.g., a combination of ABE and CBE) fused to a Cas protein. These biomolecules are named double base editors and allow editing of two different bases (Grunewald, J et al, A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing, nature Biotechnology (2020); li, C et al, targeted, random mutagenesis of plant genes with dual cytosine and adenine base editors, nature Biotechnology (2020)).

In particular embodiments, the base editing system comprises a Cytosine Base Editor (CBE) and/or an Adenosine Base Editor (ABE) as defined above.

B) As described in Anzalone, a.v. et al, nature 576,149-157 (2019), the leader editor (PE) consists of nCas9 fused to reverse transcriptase, which is fused to leader editing RNA (pegRNA; guide RNAs that include a template region for reverse transcription).

Lead editing allows for the introduction of insertions, deletions (indels) and 12 base-to-base conversions. Leader editing relies on the ability of Reverse Transcriptase (RT) fused to Cas nickase variants to convert RNA sequences brought about by leader editing guide RNA (pegRNA) into DNA at the nicking site created by the Cas protein. The DNA branch (flap) resulting from this method is then included or excluded in the targeted DNA sequence.

The pilot editing system includes:

cas nickase variants such as Cas9-H840A fused to a reverse transcriptase domain such as M-MLV RT or mutant forms thereof (M-MLV RT (D200N), M-MLV RT (D200N/L603W/T330P/T306K/W313F)

Lead editing guide RNA (pegRNA)

To facilitate editing, the lead editing system can include expression of additional sgrnas that ideally target Cas nickase activity to the non-edited DNA strand only after the edited strand branches are resolved, by designing the sgrnas to anneal to the edited strand but not to the original strand.

Non-limiting examples of lead editing systems include PE1, PE1-M2, PE1-M3, PE1-M6, PE1-M15, PE1-M3inv, PE2, PE3b.

The Cas9 reverse transcript via homolog Y is compiled in parallel ('CRISPEY'), a reverse transcriptase RNA fused with sgRNA and expressed with Cas9, and the reverse transcript protein includes at least reverse transcriptase (Sharon, e.et al, cell 175,544-557.e16 (2018)).

SCRIBE strategy: reverse transcription subsystem (Farzadfard, F. & Lu, t.k.science 346,1256272 (2014)) expressed in combination with a recombinase that promotes single-stranded DNA recombination, also known as a single-stranded annealing protein (SSAP). Such recombinases include, but are not limited to, phage recombinases such as lambda red, recET, sak, sak and Wannier, T.M. et al Improved bacterial recombineering by parallelized protein discovery.Biorxiv2020.01.14.906594 (2020) doi: 10.1101/2020.01.14.906594.

The targetron system based on group II introns is described in Karberg, m.et al, nat Biotechnol 19,1162-7 (2001), which has been adapted to many bacterial species.

Other methods of gene targeting based on reverse transcripts are described in Simon, A.J., ellington, A.D, & Finkelstein, I.J., nucleic Acids Res 47,11007-11019 (2019).

C) CRISPR/Cas in various embodiments, the sequence of interest encodes a fusion protein comprising a Cas9 (e.g., cas9 nickase) domain and a deaminase domain. In some embodiments, the fusion protein comprises Cas9 and a cytosine deaminase, such as an apodec enzyme, or an adenosine deaminase, such as an ADAT enzyme, e.g., as disclosed in U.S. patent publication 2015/0166980. In one embodiment, the deaminase is ACF1/ASE deaminase.

In various embodiments, the apodec deaminase is selected from the group consisting of an apodec 1 deaminase, an apodec 2 deaminase, an apodec 3A deaminase, an apodec 3B deaminase, an apodec 3C deaminase, an apodec 3D deaminase, an apodec 3F deaminase, an apodec 3G deaminase, and an apodec 3H deaminase. In various embodiments, the fusion protein comprises a Cas9 domain, a cytosine deaminase domain, and a Uracil Glycosylase Inhibitor (UGI) domain.

In one embodiment, the deaminase is an adenosine deaminase that deaminates adenosine in DNA, e.g., as disclosed in us patent 10,113,163. In some embodiments, the fusion protein further comprises an inhibitor of base repair, such as a nuclease-inactivated inosine-specific nuclease (dsn), e.g., as disclosed in us patent 10,113,163. In various embodiments, the nucleic acid of interest encodes a fusion protein comprising a catalytically impaired Cas9 endonuclease fused to a engineered reverse transcriptase programmed with a leader editing guide RNA (pegRNA) that both specifies the target site and encodes the desired editing, e.g., as described by Anzalone et al.

In some embodiments, other programmable nucleases can be used. These include engineered TALENs (transcription activator-like effector nucleases) and variants, engineered Zinc Finger Nuclease (ZFN) variants, natural, evolved or engineered meganuclease or recombinase variants, and any combination or hybrid of programmable nucleases. Thus, the programmable nucleases provided herein can be used to selectively modify DNA encoding a DNA sequence or gene of interest such as a toxin gene, virulence factor gene, antibiotic resistance gene, remodeling gene or regulatory gene (see WO2014124226 and US 2015/0064138).

In one embodiment, a base editing system or base editor is used to inactivate expression of a gene by editing one or several nucleotides involved in transcription or translation. More specifically, the base editor targets one or several nucleotides of the promoter, RBS or start codon.

In one embodiment, a base editing system or base editor is used to introduce a premature stop codon.

In one embodiment, a base editing system or base editor is used to introduce one or several rare codons.

In another embodiment, a base editing system or base editor is used to regulate the expression of a gene by editing one or several nucleotides involved in transcription or translation. More specifically, the base editor targets one or several nucleotides of the promoter, RBS or start codon, resulting in an increase or decrease in gene expression.

In another embodiment, a base editing system or base editor is used to restore mutations that result in inactivation, reduction, or increase in gene activity or pathway.

In another embodiment, a base editing system or base editor is used to recover mutations that result in increased pathogenicity.

In one embodiment, a base editing system or base editor is used to modify the regulation of a gene by editing one or several nucleotides involved in its regulation, such as the nucleotides of an operator sequence, a transcription factor binding site, a riboswitch, an rnase recognition site, a protease cleavage site, a methylation site, or a post-translational modification site (phosphorylation, glycosylation, acetylation, prokaryotic ubiquitination (delivery) … …).

In some embodiments, the sequence of interest encodes an RNA base editing system. RNA base editing is based on the same principle as DNA base editing: enzymes that catalyze the conversion of one RNA base to another must be in close proximity to the target base to effect its conversion locally. In one embodiment, the enzyme used for RNA editing is an adenosine deaminase from the ADAR family, which converts adenosine to inosine in the dsRNA structure. Several open studies have used this specificity for dsRNA and extended ADAR deaminase domains (ADAR _DD ) Fusion with antisense oligonucleotides to program local RNA base editing. Recently, the ability of some CRISPR-Cas systems to bind RNA molecules has been altered for RNA editing. Using a highly active mutant with ADAR2 deaminase domain (ADAR 2 for REPAIRv 1) _DD E488Q, ADAR2 for REPAIRv2 _DD The catalytic inactivation of the Cas13b enzyme (dPspCas 13 b) by E488Q-T375G fusion increases specificity and efficiency compared to previous RNA editing strategies (Cox, d.b.t. et al, science 358,1019-1027 (2017)).

Non-limiting examples of RNA base editing proteins include REPAIRv1, REPAIRv2.

In one embodiment, an RNA base editor is used to inactivate expression of a gene by editing one or several nucleotides involved in translation. More specifically, the base editor targets one or several nucleotides of the 5' utr, RBS, start codon.

In one embodiment, an RNA base editor is used to introduce a premature stop codon.

In one embodiment, an RNA base editor is used to introduce one or several rare codons.

In another embodiment, an RNA base editor is used to regulate expression of a gene by editing one or several nucleotides involved in translation. More specifically, the base editor targets one or several nucleotides of the 5' utr, RBS, start codon, resulting in an increase or decrease in gene expression.

In another embodiment, an RNA base editor is used to restore mutations that result in inactivation or reduction of the activity or pathway of a gene.

In another embodiment, a base editor is used to recover mutations that result in increased pathogenicity.

In a preferred embodiment, the sequence of interest only plays a role in the targeted bacterial cell. More preferably, the sequence of interest is expressed only in the targeted bacterial cell.

Origin of replication

In certain embodiments, the copy number of the payload in the producer bacterial cell is controlled by the at least one induction mechanism defined above. In an alternative embodiment, another induction mechanism controls the copy number of the payload in the producer bacterial cell.

Origins of replication known in the art have been derived from species-specific plasmid DNA (e.g., coIE1, rl, pT181, pSC101, pMB1, R6K, RK2, p15a, etc.), from bacterial viruses (e.g.M13, F1, and P4) and from bacterial chromosomal origins of replication (e.g., oriC).

In one embodiment, the payload used in the context of the present invention comprises a bacterial origin of replication that is functional in the targeted bacteria.

Alternatively, the payload used in the context of the present invention does not comprise any functional bacterial origin of replication or comprises an origin of replication that is inactive in the targeted bacteria. In such embodiments, the payload is unable to replicate itself once it is introduced into the bacteria by the phage particle or phage-derived delivery vehicle.

In one embodiment, the origin of replication on the payload to be packaged is inactive in the targeted bacteria, meaning that the origin of replication is not functional in bacteria targeted by the phage particle or phage-derived delivery vehicle, thereby preventing unwanted plasmid replication.

In one embodiment, the payload comprises a bacterial origin of replication that is functional in a producer bacterial cell of the invention.

Bacterial-specific origin of replication

Plasmid replication depends on host bacterial enzymes and plasmid-controlled cis-and trans-determinants. For example, some plasmids have determinants recognized in almost all gram-negative bacteria and function correctly in every host bacterium during replication initiation and regulation. Other plasmids possess this capability only in some bacteria (Kues, U and Stahl, U1989Microbiol Rev 53:491-516).

Plasmids replicate by three general mechanisms starting at the origin of replication, namely type θ, strand displacement and rolling circle (reviewed by Del Solar et al 1998Microbio and Molec Biol.Rev 62:434-464). These origins of replication contain sites required for interaction of the plasmid and/or host encoded proteins.

The origins of replication used on the payloads used in the context of the present invention may be medium copy numbers such as ColE1 ori (15-20 copies per cell) or R6K plasmids from pBR322 (15-20 copies per cell), or may be high copy numbers such as pUC oris (500-700 copies per cell), pGEM oris (300-400 copies per cell), pTZ oris (> 1000 copies per cell) or pBluescript oris (300-500 copies per cell).

In one embodiment of the present invention, in one embodiment, bacterial origins of replication are selected from the group consisting of ColE1, pMB1 and variants (pBR 322, pET, pUC, etc.), P15a, colA, colE2, pOSAK, pSC101, R6K, incW (pSa, etc.), incFII, pT181, P1, F IncP, incC, incJ, incN, incP, incP4, incQ, incH11, RSF1010, cloDF13, NTP16, R1, F5, pPS10, pC194, pE194, BBR1, pBC1, pEP2, pWVO1, pLF1311, pAP1, pWKS1, pLS11, pUB6060, pJDMb 4, pJ 101, pSN22, pAeta 1, pIP501, pIP407, ZM6100 (Sa 1, RA3, pMOL98, RK2/RP4/RP1/R68, pB10, R300B, pRO 4, pRO1600, pEM 2, pA 84, pFL 3, pFL 80, pFv 387, pFv 80 and pU 9.

More preferably, the bacterial origin of replication is an E.coli origin of replication selected from the group consisting of: colE1, pMB1 and variants (pBR 322, pET, pUC, etc.), P15a, colA, colE2, pOSAK, pSC101, R6K, incW (pSa, etc.), incFII, pT181, P1, F IncP, incC, incJ, incN, incP1, incP4, incQ, incH11, RSF1010, cloDF13, NTP16, R1, F5, pPS10.

More preferably, the bacterial origin of replication is selected from the group consisting of pC194, pE194, BBR1, pBC1, pEP2, pWVO1, pLF1311, pAP1, pWKS1, pLS11, pUB6060, pJD4, pIJ101, pSN22, pAMbreta 1, pIP501, pIP407, ZM6100 (Sa), pCU1, RA3, pMOL98, RK2/RP4/RP1/R68, pB10, R300B, pRO1614, pRO1600, pECB2, pCM1, pFA3, repFIA, repFIB, repFIC, pYVE439-80, R387, phasyl, RA1, TF-FC2, pMV158 and pUB113.

Even more preferably, the bacterial origins of replication are ColE1 and p15a.

In one embodiment, the bacterial origin of replication is functional in Propionibacterium (Propionibacterium) and dermatophytes (Cutibacillus), more specifically Propionibacterium freudenreichii (Propionibacterium freudenreichii) and dermatophytes acnes, and is selected from the group consisting of pLME108, pLME106, p545, pRGO1, pZGX01, pPG01, pYS1, FRJS12-3, FRJS25-1, pIMPLE-HL096PA1, A_15_1_R1. In a particular embodiment, the bacterial origin of replication is selected from the bacterial origins of replication disclosed in U.S. application Ser. No. 2022/135986 and U.S. application Ser. No. 2022/135787.

Phage origin of replication

Payloads used in the context of the present invention may comprise phage origins of replication that may be accompanied by complementation of the complete phage genome to initiate replication of the payload for later encapsulation into a different capsid.

Phage origins of replication can also be engineered to act as bacterial origins of replication without the need to package any phage particles.

The phage origin of replication contained in the payload used in the context of the present invention may be any origin of replication found in phage.

Preferably, the phage origin of replication is M13, f1,Wild-type or non-wild-type sequences of P4, λ, P2, 186, λ -like, HK022, mEP237, HK97, HK629, HK630, mEP043, mEP213, mEP234, mEP390, mEP460, mpex 1, mEPx2, phi80, mEP234, T2, T4, T5, T7, RB49, phiX174, R17, PRD1 Pl-like, P2-like, P22-like, N15 and N15-like bacteriophage.

More preferably, the phage origin of replication is selected from M13, f1,Phage origin of replication of P4 and lambda.

In a particular embodiment, the phage origin of replication is a P4 origin of replication.

In a particular embodiment, the phage origin of replication is from a propionibacterium phage: BW-like phages such as doucete, B22, E6, G4; BV-like phages such as anagle, E1, B3; BX-like phages such as PFR1 and PFR2; filamentous B5 phage; BU-like phage (acnes skin bacillary phage). In a particular embodiment, the phage origin of replication is selected from the phage origins of replication disclosed in U.S. application Ser. No. 2022/135986 and U.S. application Ser. No. 2022/135987, incorporated herein by reference.

Conditional origin of replication

In certain embodiments, the payload comprises a conditional origin of replication that is inactive in the targeted bacteria, but active in the producing bacterial cells.

In the context of the present invention, a "conditional origin of replication" refers to an origin of replication whose functionality can be controlled by the presence of a particular molecule.

In particular embodiments, the conditional origin of replication is an origin of replication, the replication of which is dependent on the presence of one or more given proteins, peptides, RNAs, nucleic acids, molecules or any combination thereof.

In certain embodiments, replication involving the origin of replication may be further dependent on a process such as transcription to activate the replication.

In the context of the present invention, the conditional origin of replication is inactive in the targeted bacteria, since the given protein, peptide, RNA, nucleic acid, molecule or any combination thereof is not present in the targeted bacteria.

In certain embodiments, the conditional origin of replication is active in the producer bacterial cell because the producer bacterial cell expresses the given protein, peptide, RNA, nucleic acid, molecule, or any combination thereof. In certain embodiments, the protein, peptide, RNA, nucleic acid, molecule, or any combination thereof is expressed in trans in the producer bacterial cell.

By "trans" is meant herein that the protein, peptide, RNA, nucleic acid, molecule, or any combination thereof is not encoded on the same nucleic acid molecule as the nucleic acid molecule comprising the origin of replication. In a particular embodiment, the protein, peptide, RNA, nucleic acid, molecule or any combination thereof is encoded on a chromosome or a vector, in particular a plasmid. In certain embodiments, the vector comprises an antibiotic resistance marker. In an alternative embodiment, the vector is free of an antibiotic resistance marker.

Because the conditional origin of replication is inactive in the targeted bacteria due to the absence of the given protein, peptide, RNA, nucleic acid, molecule, or any combination thereof in the targeted bacteria, the conditional origin of replication may be selected according to the particular bacteria to be targeted.

The conditional replication origin disclosed herein may originate from a plasmid, a bacteriophage or a PICI, which preferably shares the following features: they contain a repeat sequence or a repeat in their origin of replication and they encode at least one protein (i.e. Rep, protein O, protein P, pri) that interacts with the origin of replication specific for them.

For example, mention may be made of the following conditional replication systems of plasmids and bacteriophages: RK2, R1, pSC101, F, rts1, RSF1010, P1, P4, lambda, phi82, phi80.

In particular embodiments, the conditional origin of replication is selected from the group consisting of an R6K lambda DNA origin of replication and derivatives thereof, an IncP alpha oriV origin of replication and derivatives thereof, a ColE1 origin of replication modified to be under an inducible promoter, and an origin of replication from a phage-inducible chromosomal island (PICI) and derivatives thereof.

In particular embodiments, the conditional origin of replication is an origin of replication present in less than 50% or less than 40%, less than 30%, less than 20%, less than 10% or less than 5% of the microbiome bacteria of the host subject.

In another specific embodiment, the conditional origin of replication comprises or consists of: replication origin sequences which are less than 80%, in particular less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5% or less than 1% identical to the replication origin sequences of a bacterium of the microbiome of the host subject, in particular representing more than 50%, more than 70%, more than 80%, more than 90% or more than 95% of the bacteria of the microbiome of the host subject.

As used herein, the term "phage-inducible chromosomal island" or "PICI" refers to a mobile genetic element having conserved genetic organization and encoding a pair of different regulatory genes, including the PICI major repressor. Generally, in gram-positive bacteria, PICI transcribed to the left of rpr and in the same direction encodes a small set of genes, including the integrase (int) gene; to the right of rpr and transcribed in the opposite direction, the PICI encodes a excision function (xis) and a replication module consisting of a primase homolog (pri) and optionally a replication initiator (rep) sometimes fused together, followed by an origin of replication (ori); PICI, which is next to these genes and also transcribed in the same direction, encodes genes involved in phage interference and optionally terminates small subunit homologs (terS).

In a specific embodiment, the conditional origin of replication is an origin of replication derived from a phage-induced chromosomal island (PICI).

The specific conditional origin of replication is indeed derived from the PICI.

It was shown that it is possible to derive novel conditional replication vectors, in particular based on the primase-helicase and the origin of replication from PICI. These origins can be relatively rare in the target strain, and more advantageously the priming enzyme-ori pair can be unique for each PICI, significantly reducing the likelihood of undesired recombination or payload diffusion events. They may be further modified to further limit the chance of recombination and to remove restriction sites to bypass the defense system of the target bacteria.

In a particular embodiment, the conditional replication origin is derived from The replication origin of PICI from Escherichia coli strain CFT073, which is disclosed in Fillol-Salom et al, (2018) The ISME Journal 12:2114-2128.

In a particular embodiment, the conditional origin of replication is the primase ori of PICI from E.coli strain CFT073, typically having the sequence SEQ ID NO. 1.

In another particular embodiment, the conditional origin of replication is the priming enzyme ori of PICI from escherichia coli strain CFT073, comprising no at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, or at least 16 restriction sites selected from the group consisting of: GAAABCC, GCCGGC, RCCGGY, GCNGC, TWCANNNNNNTGG (SEQ ID NO: 2), TGGCCA, ACCYAC, YGGCCR, AGACC, GCWGC, GGGANGC, GKAGATD, GCCGGYYD, GGCYAC, RGCCGGYYD and VGCCGGYBD.

In a particular embodiment, the conditional origin of replication is the primase ori of PICI from E.coli strain CFT073, without the restriction site GAAABCC. Preferably, the conditional origin of replication has the sequence SEQ ID NO. 3.

In another particular embodiment, the conditional origin of replication is the priming enzyme ori of PICI from E.coli strain CFT073, free of restriction sites GAAABCC, GCCGGC, RCCGGY, GCNGC, TWCANNNNNNTGG (SEQ ID NO: 2), TGGCCA, ACCYAC, YGGCCR, AGACC, GCWGC, GGGANGC, GKAGATD, GCCGGYYD, GGCYAC, RGCCGGYYD and

VGCCGGYBD. Preferably, the conditional origin of replication has the sequence SEQ ID NO. 4.

In a specific embodiment, wherein said origin of replication is derived from a phage-induced chromosomal island (PICI), said conditional origin of replication is active in said producer bacterial cell, because said producer bacterial cell expresses a rep protein, in particular a primase-helicase of sequence SEQ ID NO:5, typically encoded by a nucleic acid comprising or consisting of sequence SEQ ID NO: 6.

These specific conditional origins of replication proved to be particularly compatible with lambda-based packaging, resulting in sufficiently high titers required for microbiota-related applications >10 ¹⁰ /mL)。

Preferably, the producer bacterial cells stably contain the payload and are capable of replicating the payload.

In particular embodiments, the donor bacterial cell expresses a given protein, peptide, nucleic acid, RNA, molecule, or any combination thereof when the conditional origin of replication of the payload is one whose replication depends on the presence of the protein, peptide, nucleic acid, RNA, molecule, or any combination thereof. Preferably, the protein, peptide, nucleic acid, RNA, molecule or any combination thereof is expressed in trans, as defined above.

In particular embodiments, the producer bacterial cell stably comprises a nucleic acid encoding the protein, peptide, nucleic acid, RNA, molecule, or any combination thereof.

In a specific embodiment, when the origin of replication is derived from a phage-induced chromosomal island (PICI), the conditional origin of replication is active in the donor bacterial cell, because the donor bacterial cell expresses a rep protein, in particular a primase-helicase of sequence SEQ ID NO: 5.

In a particular embodiment, the producer bacterial cell stably comprises a nucleic acid encoding the rep protein, in particular the primase-helicase, which nucleic acid generally comprises or consists of the sequence SEQ ID NO. 6.

Packaging site

In a particular embodiment, the payload is a nucleic acid payload comprising a packaging site derived from the lytic bacteriophage.

"packaging site" means a phage baseGenome packaging into viral particles. Host-specific bacteriophages (and packaging sites therefor) include, but are not limited to, SPP1 (SPP 1 pac site), P1 (P1 pac site), T1 (T1 pac site), T7 (T7 concatemeric), lambda (cos site), mu (mu pac site), P22 (P22 pac site), and,(/>pac site), sf6 (Sf 6 pac site), 149 (149 pac site) and a1122 (a 1122-concatemer junction). For most bacteriophages, the packaging site is called the pac site. In some cases, the packaging site is referred to as a concatemer junction (e.g., a T7 concatemer junction). In each case, the packaging site is substantially separated from the naturally occurring sequences adjacent thereto in the bacteriophage genome.

For some bacteriophages, the packaging site may be unknown. In these cases, the pac site can be determined by the nature of the plasmid being packaged with the functional bacteriophage pac site. For example, the DNA sequence required for lambda packaging of a bacteriophage is determined by incorporating a small restriction fragment of the genomic DNA of the lambda bacteriophage into a plasmid (Hohn 1983PNAS USA 80:7456-7460). After introduction into the in vivo packaging strain, the efficiency of packaging/transduction was quantitatively assessed. Using similar strategies, the pac sites of many bacteriophages were determined: lambda (Miwa 1982Gene 20:267-279); mu (Croenen et al, 1985Virology 144:520-522); filamentous bacteriophage, including f1, fd, M13, and Ike (Russel et al, 1989J Virol 1989 63:3284-3295); p22 (Petri et al, 1990Gene88:47-55; wu et al, 2002MolecMicrobiol 45:1631-1646); t7 (Chung et al 1990J Mol Biol 216:927-938) and T3 (Hashimoto et al 1992Virology 187:788-795).

In particular embodiments, the packaging sites are as disclosed in U.S. applications US2022/135986 and US 2022/135987.

Other components of the payload

The payloads used in the context of the present invention are preferably devoid of antibiotic resistance markers.

Antibiotic resistance genes are well known in the art and include, but are not limited to, ampicillin Lin Kangxing (Amp), chloramphenicol resistance (Cm), tetracycline resistance (Tet), kanamycin resistance (Kan), hygromycin resistance (Qiyg or hph genes), and zeomycin resistance (Zeo).

In certain embodiments, the payloads used in the context of the present invention comprise an auxotrophic marker. Auxotrophic markers in bacteria have previously been described, for example, in U.S. Pat. nos. 4,920,048, 5,691,185, 6,291,245, 6,413,768, and 6,752,994; U.S. patent publication No. 20050186666; struhl et al, (1976) PNAS USA 73;1471-1475; macCormick et al, (1995) FEMS Microbiol. Lett.127:105-109; dickely et al, (1995) mol. Microbiol.15:839-847; sorensen et al, (2000) appl. Environ. Microbiol 66:1253-1258; and Fiedler & Skerra (2001) Gene 274:111 118, and generally includes DapA and ThyA. In a particular embodiment, the auxotrophic marker is ThyA.

In certain embodiments, the payload does not comprise any restriction sites that are frequently recognized by restriction enzymes encoded by the targeted bacterial cells. In another particular embodiment, the payload comprises no more than 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 restriction sites that are frequently recognized by a restriction enzyme encoded by the targeted bacterial cell or group or set of targeted bacterial cells.

As used herein, the term "Restriction sites"He"Restriction enzyme site"equivalent" and refers to a position on a nucleic acid that contains a particular nucleotide sequence that is recognized by a restriction enzyme. In particular, nucleic acids comprise specific sequences that are bound and cleaved by restriction enzymes. The restriction site is typically a palindromic sequence of 4-8 base pairs in length. More precisely, a restriction site refers to a specific sequence and modification state so as to be bound and cleaved by a restriction enzyme. In particular, it refers to a specific unmodified sequence for binding and cleavage by restriction enzymes. In particular the sequence is free of methylation, methylolation and glucosyl-hydroxylsMethylation. In this case, the restriction enzyme belongs to I, II or III. Alternatively, it may refer to a specific modified sequence so as to be bound and cleaved by restriction enzymes, such as methylated, methylolated and glucosyl-methylolated DNA. In this case, the restriction enzyme belongs to type IV.

As used herein, "with respect to restriction sites and restriction enzymes"Identified by …"means that the restriction site is cut by a restriction enzyme.

In the restriction site sequence, N means that the nucleotide may be A, C, G or T; b means that the nucleotide may be C, G or T; y means that the nucleotide may be C or T; w means that the nucleotide may be a or T; r means that the nucleotide may be A or G; and D means A, G or T.

As used herein, the term "Restriction enzymes"He"Restriction endonucleases"equivalent" and refers to an enzyme that cleaves nucleic acid at or near a restriction site. Restriction enzymes are generally classified into four types (type I-type IV). The REBASE database allows the listing of restriction sites that can be recognized by a given bacterium based on the restriction enzyme expressed by it.

In the group of bacteria of interest, "Often timesBy "or" frequently "is meant that at least 10, 20, 30, 40, 50, 60, 70, 75, 80, 85, 90, 95, or 99% of the bacteria in the group encode a restriction enzyme.

The payload according to the invention preferably comprises no more than 100 restriction sites. In a preferred embodiment, the payload according to the invention comprises no more than 10 restriction sites. In a most preferred embodiment, the payload according to the invention does not comprise any restriction sites.

Targeting bacteria

The bacteria targeted by the phage particles or phage-derived delivery particles of the invention can be any bacteria present in a mammalian organism, plant or environment. It may be any commensal, commensal or pathogenic bacteria of the microbiota or microbiome.

The microbiome can comprise a variety of endogenous bacterial species, any of which can be targeted in accordance with the present disclosure. In some embodiments, the genus and/or species targeted to the endogenous bacterial cell may depend on the first type of bacteriophage as defined in the section "bacteriophage and genes derived from bacteriophage" above. For example, some bacteriophage exhibit tropism for or preferentially target a particular host bacterial species. Other bacteriophages do not exhibit such tropism and can be used to target endogenous bacterial cells of many different genera and/or species.

Examples of bacterial cells include, without limitation, cells from bacteria of the following genera: yersinia species (Yersinia spp.), escherichia species (Escherichia spp.), klebsiella species (Klebsiella spp.), acinetobacter species (Acinetobacter spp.), bolde genus species (Bordetella spp.), neisseria species (Neisseria spp.), aeromonas species (Aeromonas spp.), francisella species (Francisella spp.), corynebacterium species (Corynebacterium spp.), citrobacter species (Citrobacter spp.), chlamydia species (Chlamydia spp.), hamammitis species (Hamammitis spp.), haemophilus species (Herophila spp.), brucella spp.), bacillus species (Mycobacterium spp.), legionella species (Legionella spp.). Rhodococcus species (Rhodococcus spp.), pseudomonas species (Pseudomonas spp.), helicobacter species (Helicobacter spp.), vibrio species (Vibrio spp.), bacillus species (Bacillus spp.), erysipelas species (Erysipelothrix spp.), salmonella species (Salmonella spp.), streptomyces species (Streptomyces spp.), streptococcus species (Streptococcus spp.), rhodobacter spp Staphylococcus species (Staphylococcus spp.), bacteroides species (Bacteroides spp.), prasugrel species (Prevotella spp.), clostridium species (Clostridium spp.), bifidobacterium species (Bifidobacterium spp.), clostridium species, brevibacterium species (breve bacterium spp.), lactococcus species (Lactococcus spp.), and the like, leuconostoc species (Leuconostoc spp.), actinobacillus species (actionicos spp.), selenomonas species (selnomomonas spp.), shigella species (Shigella spp.), zymomonas species (zymomonas spp.), mycoplasma species (myccoplasma spp.), treponema spp.), leuconostoc species, corynebacterium species, enterococcus species (Enterococcus spp.), enterobacter spp), enterobacter species (Enterobacter spp.), pyrococcus spp), pyrococcus species (Pyrococcus spp.), serratia species (Serratia spp.), morganella species (Morganella spp), morganella species (Parvinus spp), clostridium species (Fusobacterium spp.), fusobacterium species (Furacius spp) actinomycete species (Actinomyces spp.), porphyrinomonas species (Porphyromonas spp.), micrococcus species (Micrococcus spp.), bartonella spp.), borrelia species (Borrelia spp.), brucella species (Brucella spp.), campylobacter species (Campylobacter spp.), chlamydophila spp.), chlamydia species (Chlamydophila spp.), dermatophyta species (Cutibacterium spp.), propionibacterium species (Propionibacterium spp.), gardnerella spp.), ehrlica species (Ehrlichia spp.), haemophilus species (Haemophilus spp), leptospira species (Leospora spp.), listeria species (Listeria spp.), mycobacterium spp, nocardia spp, rickettsia spp, ureaplasma spp, lactobacillus spp, faecalis spp, ruminococcus spp, and mixtures thereof.

Thus, phage particles, phage delivery particles, and/or phage may target (e.g., specifically target) bacterial cells from any one or more of the above bacterial genera, particularly to specifically deliver a payload according to the invention.

Preferably, the targeting bacteria may be selected from the group consisting of yersinia species, escherichia species, klebsiella species, acinetobacter species, pseudomonas species, helicobacter species, vibrio species, salmonella species, streptococcus species, staphylococcus species, bacteroides species, clostridium species, shigella species, enterococcus species, enterobacter species, listeria species, dermatophytes species, propionibacterium species, fusobacterium species, porphyromonas species, and gardnerella species.

In some embodiments, the targeted bacteria are anaerobic bacterial cells (e.g., cells that do not require oxygen for growth). Anaerobic bacterial cells include facultative anaerobic cells such as, but not limited to, escherichia coli, osnescentella (Shewanella oneidensis), gardnerella vaginalis (Gardnerella vaginalis), and listeria. Anaerobic bacterial cells also include obligate anaerobic cells such as bacteroides, clostridia, dermatophytes, propionibacteria, fusobacterium and porphyromonas species. Anaerobic bacteria are most commonly found in the gastrointestinal tract in humans. In some particular embodiments, the targeted bacteria are thus the bacteria most commonly found in the gastrointestinal tract. The bacteriophage and subsequent phage particles, phage delivery vehicles, and/or phages used to prepare the hybrid helper phage may target (e.g., specifically target) anaerobic bacterial cells according to their specificity profile known to those of skill in the art to specifically deliver plasmids.

In some embodiments, the targeted bacterial cells are, without limitation, bacteroides thetaiotaomicron (Bacteroides thetaiotaomicron), bacteroides fragilis, bacteroides dirachta (Bacteroides distasonis), bacteroides vulgare (Bacteroides vulgatus), clostridium tenella (Clostridium leptum), clostridium globosum (Clostridium coccoides), staphylococcus aureus, bacillus subtilis (Bacillus subtilis), clostridium butyricum (Clostridium butyricum), brevibacterium lactofermentum (Brevibacterium lactofermentum), streptococcus agalactiae (Streptococcus agalactiae), lactobacillus lactiae (Lactococcus lactis), leuconostoc lactis (Leuconostoc lactis), actinobacillus (Actinobacillus actinobycetemcomitans), cyanobacteria (cyanobacteria), escherichia coli, helicobacter pylori (Helicobacter pylori), selenomonas ruminants (Selnomonas ruminatium), shigella sonneisseria sonnei (Shigella sonnei), zymomonas mobilis (Zymomonas mobilis), mycoplasma (Mycoplasma mycoides), streptococcus (Treponema denticola), bacillus thuringiensis (Bacillus thuringiensis), staphylococcus (Staphilococcus lugdunensis), leuconostoc (Leuconostoc oenos), corynebacterium xerosis (Leuconostoc oenos), lactobacillus acidophilus (Leuconostoc oenos), lactobacillus (Leuconostoc oenos) and lactobacillus (Leuconostoc oenos), synechocystis (Synechocystis) strain PCC6803, bacillus liquefaciens (Bacillus liquefaciens), bacillus pumilus (Pyrococcus abyssi), zygomonas ruminant (Selenomonas nominantium), lactobacillus johnsonii (Lactobacillus hilgardii), streptococcus wildens (Streptococcus ferus), lactobacillus pentosus (Lactobacillus pentosus), bacteroides fragilis, staphylococcus epidermidis (Staphylococcus epidermidis), streptomyces secretes (Streptomyces phaechromogenes), streptomyces ganensis (Streptomyces ghanaenis), klebsiella pneumoniae (Porphyromonas endodontalis), enterobacter cloacae (Enterobacter cloacae), enterobacter aerogenes (Enterobacter aerogenes), serratia marcescens (Serratia marcescens), morganella morganii (Morganella morganii), citrobacter freundii (Citrobacter freundii), propionibacterium freudenreichii, pseudomonas aeruginosa, micromonospora parvula (Parvimona micra), primeria intermedia (Prevotella intermedia), fusarium nucleatum, proteus melanogaster (Prevotella nigrescens), serratia pseudolaris (Actinomyces israelii), porphyromonas (Porphyromonas endodontalis), porphyromonas gingivalis (Porphyromonas endodontalis), micrococcus flavus (Porphyromonas endodontalis), bacillus sphaericus (Porphyromonas endodontalis), bordetella (Porphyromonas endodontalis) and Bordetella (Porphyromonas endodontalis) are obtained by the methods of producing the strain, borrelia albopicta (Borrelia afzeli), borrelia febrile (Borrelia recurrentis), brucella abortus (Brucella abortus), brucella canis (Brucella canis), brucella ovis (Brucella melitensis), brucella suis (Brucella suis), campylobacter jejuni, campylobacter coli (Campylobacter coli), campylobacter foetidus (Campylobacter fetus), chlamydia pneumoniae (3835), chlamydia trachomatis (Chlamydia trachomatis), chlamydophila psilosis (Chlamydophila psittaci), clostridium botulinum (Clostridium botulinum), clostridium difficile (Clostridium difficile), clostridium perfringens (Clostridium perfringens), clostridium tetani (Clostridium tetani), corynebacterium diphtheriae (Corynebacterium diphtheria), acnes (formerly propionibacterium acnes (Propionibacterium acnes), canine Ehrlichia (Ehrlichia canis), escherichia coli (Ehrlichia chaffeensis), enterococcus faecium (Enterococcus faecium), francisco, haemophilus influenzae (Haemophilus influenza), pneumophila (Sang Daluo), leptospira (Sang Daluo) and leptospira (Sang Daluo, neisseria gonorrhoeae (Neisseria gonorrhoeae), neisseria meningitidis, nocardia astronomy (Nocardia asteroids), rickettsia (Rickettsia rickettsia), salmonella enteritidis (Salmonella enteritidis), salmonella typhi (Salmonella typhi), salmonella paratyphi (Salmonella paratyphi), salmonella typhimurium (Salmonella typhimurium), shigella flexneri (Shigella flexnerii), shigella dysenteriae, staphylococcus saprophyticus (Staphylococcus saprophyticus), streptococcus pneumoniae (Streptococcus pneumoniae), streptococcus pyogenes, gardnerella vaginalis, streptococcus viridis (Streptococcus viridans), treponema pallidum (Treponema pallidum), ureaplasma urealyticum (Ureaplasma urealyticum), vibrio cholerae, vibrio parahaemolyticus (Vibrio parahaemolyticus), yersinia pestis, yersinia enterocolitica (Yersinia enterocolitica), yersinia pseudotuberculosis (Yersinia pseudotuberculosis), actinobacillus baumannii (Actinobacter baumanii), pseudomonas aeruginosa and mixtures thereof, preferably the bacteria of interest are selected from the group consisting of escherichia coli, enterococcus faecium, staphylococcus aureus, staphylococcus pneumoniae, klebsiella pneumoniae (Acinetobacter baumanii), pseudomonas baumannii and mixtures thereof.

In some embodiments of the present invention, in some embodiments, the targeted bacterial cells are, without limitation, anaerobic bacteria (Anaeroruncus), acetobacter (Acetarobacterium), polyacetobacter (Acetamaculococcus), acetobacter (Acetavibrio), anaerobic coccus (Anaerocrocus), anaerobic bacteria (Anaeroaeroaeroaerofelium), anaerobic campylobacter (Anaerosporus), anaerobic bacteria (Anaerosporidium), anaerobic bacteria (Anaerosporium), anaerobic bacteria (anasporax), butyric acid bacteria (Vibrio butyricum), clostridium, faecal bacteria (Capracocucus), dehalobacter (Dehalobacter), dai A listeria (dialiter), dunaliella (Dorea), enterococcus (Entercoccus), ethanol producing bacteria (ethanol) bacteria), faecalis, clostridium, gracilomyces (Gracilomyces), and Thermomyces (Dehalobacter) the genus Gugium (Gugigeheimia), the genus Proprietaria (Hesperlla), the genus Mucor (Lachnobabacterium), the genus Mucor (Lachnospira), the genus Lactobacillus, the genus Leuconostoc, the genus Megamonas (Megamonas), the genus Mo Lishi (Morella), the genus Albaea (Mitsuokella), the genus Klebsiella (Oribacterium), the genus Acetobacter (Oxobabacter), the genus Papilobabacterium (Papilibacter), the genus Proprinimira, the genus Vibrio pseudobutyric (Pseudomonas), the genus Pseudomonas (Pseudomonas), the genus Luo Sibai (Roseburia), the genus Rumezococcus, the genus Sarcina (Sarcina), the genus Serratia (Serratia), the genus Volum (Shonikowiella), the genus Sporothia (Sporothia), the genus Sporothrina (Sporothia), spore bacteria (Sporobacterium), streptococcus, rare small coccus (Subdelloglycus), camping coccus (Synthophorcoccus), thermotolerant bacillus (Thermobacillus), zymobacter (Turibacter), weissella (Weissella), clostridium, bacteroides, ruminococcus, thermus, treponema, kochia (Phascobactirium), megasphaera, thermus, bifidobacterium, lactobacillus, saspecific bacteria (Sutterella) and/or Prevotella.

In other embodiments, the targeted bacterial cells are, without limitation, achromobacter xylosoxidans (Achromobacter xylosoxidans), achromobacter fermentum (Acidaminococcus fermentans), acidococcus enterolyticus (Acidaminococcus intestini), acidococcus species, acinetobacter baumannii, acinetobacter agaragar (Acinetobacter junii), acinetobacter lofei (Acinetobacter lwoffii), actinobacillus capsulatus (Actinobacillus capsulatus), actinobacillus naeslundii (Actinomyces naeslundii), actinobacillus neotami (Actinomyces neuii), actinobacillus lycescens (Actinomyces odontolyticus), ra Ding Fangxian (Actinomyces radingae), aldeller oxydans (Adlercreutzia equolifaciens), microbacterium malasii (Aeromicrobium massiliense), actinobacillus actinobacillus (Aggregatibacter actinomycetemcomitans), acinetobacter mucin (Akkermansia muciniphila), acidovorax oceanicus (Aliagarivorans marinus), acinetobacter fena (Alistipes finegoldii), flavobacterium shikim (Alistipes indistinctus), alisti (Alistis inogenes), alistias inogenes, altissimi (Alistipes onderdonkii), acinetobacter aiensis (Alistipes onderdonkii), anemophilus (Alistipes onderdonkii), anaerobiosae (Alistipes onderdonkii), leucopia (Paeder) and Metropsystem (Alistipes onderdonkii), human anaerobic coryneform bacterium (Anaerofustis stercorihominis), anaerobic coryneform bacterium (Anaerostipes caccae), anaerobic coryneform bacterium (Anaerostipes hadrus), anaerobic coryneform bacterium (Anaerotruncus colihominis), thioflavine Jie Liuan Bacillus (Aneurinibacillus aneurinilyticus), bacillus licheniformis (Bacillus licheniformis), bacillus anorexia mosaic (Bacillus naseioianoxius), bacillus mosaic (Bacillus massiliosenegalensis), bacillus simplex (Bacillus simplex), bacillus smithii (Bacillus smithii), bacillus subtilis, bacillus thuringiensis, bacillus caldanus (Bacillus timonensis), xylanolytic pseudobacillus (bacteroides, bacteroides acidophilus (Bacteroides acidifaciens) Bacteroides (Bacteroides caccae), bacteroides multocida (Bacteroides capillosus), bacteroides thetaiotaomicron (Bacteroides cellulosilyticus), klaugus bacteroides (Bacteroides clarus), bacteroides pteronyssinus (Bacteroides coprocola), bacteroides faecalis (Bacteroides coprophilus), bacteroides dorsalis (Bacteroides coprophilus), bacteroides elvan (Bacteroides coprophilus), bacteroides pteronyssinus (Bacteroides coprophilus), bacteroides fragilis, bacteroides gallinarum (Bacteroides coprophilus), bacteroides enterobacteroides enteroides (Bacteroides coprophilus), bacteroides nori (Bacteroides coprophilus), bacteroides oleaginous (Bacteroides coprophilus), bacteroides ovatus (Bacteroides coprophilus), bacteroides pectophilum (Bacteroides coprophilus), bacteroides pini (Bacteroides coprophilus), bacteroides salvinsis (Bacteroides salanitronis), bacteroides salvinsis (Bacteroides salyersiae), bacteroides species (Bacteroides sp.), bacteroides faecalis (Bacteroides stercoris), bacteroides thetaiotaomicron, bacteroides simplex (Bacteroides uniformis), bacteroides vulgaris, bacteroides xylan, bacteroides pectophilum (Bilophila wadsworthia), human intestinal barren's bacterium (Barnesiella intestinihominis), bavariella salicina (Bavariicoccus seileri), bifidobacterium adolescentis (Bifidobacterium adolescentis), bifidobacterium angulatus (Bifidobacterium angulatum), bifidobacterium animalis (Bifidobacterium animalis), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium breve (Bifidobacterium breve), bifidobacterium minor (Bifidobacterium catenulatum), bifidobacterium denticola (Bifidobacterium dentium), bifidobacterium hyperlupulum (Bifidobacterium gallicum), bifidobacterium longum (Bifidobacterium longum), bifidobacterium pseudocatenulatum (Bifidobacterium pseudocatenulatum), bifidobacterium faecalis (Bifidobacterium stercoris), cholangium (Bilophila wadsworthia), brucella faecalis (baubaiensis), brucella hanensis (Blbaumannii), bifidobacterium hydrogenotrophic (Blautia hydrogenotrophica), brucella baudiana (Blbauxitata), blbaudiana (Blbauxites) and Blbaumannii (54-3), blbaudiana (Blbaudiana, blbaudiana (Blbaumannii), blbaumannii (Bjoba, blbaumannii) and Blbaumannii (Blbaudiana) to produce Brua (Bruki) and Blbaudiana (Blbaudiana) from these species (Blbaudiana) to produce Brua (Brufii.sp., white diarrhea butyric acid coccus (Butyricicoccus pullicaecorum), viral butyric acid unicona (Butyricimonas virosa), ear of the species vibrio (Butyrivibrio crossotus), fibrinolytic butyric acid vibrio (Butyrivibrio fibrisolvens), calcium faecalis (Caldicoprobacter faecalis), campylobacter conciseness (Campylobacter concisus), campylobacter jejuni, campylobacter Uppsala (Campylobacter upsaliensis), streptococcus light (Catenibacterium mitsuokai), ceticillium dycepacia davisae (cedea davidiana), mosaic (Cellulomonas massiliensis), whale soxhlet (Cetobacterium somerae), bergamot (Cetobacterium somerae), citrobacter freundii, bergamot (Cetobacterium somerae), citrobacter species (Citrobacter sp.), bergamot (Cetobacterium somerae) clostridium avium (Cetobacterium somerae), clostridium order (clostridium bacteria), clostridium difficile (Cetobacterium somerae), clostridium asparagicum (Cetobacterium somerae), clostridium bartrefoil (Cetobacterium somerae), clostridium bovinii (Cetobacterium somerae), clostridium halinensis (Cetobacterium somerae), clostridium hainanensis (Cetobacterium somerae), clostridium tenella (Cetobacterium somerae), clostridium methylpentosum (Cetobacterium somerae), clostridium perfringens (Cetobacterium somerae), clostridium thermocellum (Cetobacterium somerae), clostridium multi-branch (Cetobacterium somerae), clostridium scinticum (Cetobacterium somerae), clostridium species, clostridium spirans (Cetobacterium somerae), clostridium sporogenes (Cetobacterium somerae), clostridium symbiotic (Clostridium symbiosum), clostridium aerogenes (Clostridium symbiosum), colibacillus enterogenes (Clostridium symbiosum), colibacillus faecalis (Clostridium symbiosum), colibacillus in the field (Clostridium symbiosum), bacillus moniliformis (Clostridium symbiosum), bacillus fastidious (Clostridium symbiosum), clostridium dexterium (Clostridium symbiosum), enterococcus faecalis (Clostridium symbiosum), corynebacterium ammoniagenes (Clostridium symbiosum), corynebacterium aceti (Clostridium symbiosum), corynebacterium pseudodiphtheriae (Clostridium symbiosum), dermatophytes acnes, dermatophytes (Clostridium symbiosum), hafnia sulfenas (Clostridium symbiosum), vibrio fei (Clostridium symbiosum), vibrio lazii (Clostridium symbiosum), listeria succinogenes (Clostridium symbiosum), dipivoxil (Clostridium symbiosum), docarum methanoicum (Clostridium symbiosum), polyfertilus (Clostridium symbiosum), polyfern (Clostridium symbiosum), campylobacter oxydans (Clostridium symbiosum), pseudomonas putida (Clostridium symbiosum), escherichia coli (Clostridium symbiosum) and escherichia coli (Clostridium symbiosum), enterobacter cloacae, enterobacter Marseis (Enterobacter massiliensis), enterobacter lead (Enterococcus casseliflavus), enterobacter durans (Enterococcus durans), enterococcus faecium, enterococcus flavus (Enterococcus flavescens), enterococcus gallinarum (Enterococcus gallinarum), enterobacter species (Enterococcus sp.), vibrio melanogaster (Enterovibrio nigricans), leptococcus polymyxa (Erysipelatoclostridium ramosum), escherichia coli, escherichia species (Escherichia sp.), eubacterium bifidum (Eubacterium biforme), eubacterium elongatum (Eubacterium dolichum), eubacterium cholerae (Eubacterium hallii), eubacterium mucilaginosum (Eubacterium hallii), eubacterium fine (Eubacterium hallii), eubacterium rectum (Eubacterium hallii), eubacterium inert, eubacterium avium (Eubacterium hallii), hai (Eubacterium hallii), microbacterium water (Eubacterium hallii), bacillus faecalis (Eubacterium hallii), bacillus coagulans (Eubacterium hallii), protoxillus (Eubacterium hallii), bacillus sp (Eubacterium hallii), funeisseria (Eubacterium hallii), fusobacterium (Eubacterium hallii) and Fusobacterium (Eubacterium hallii) are considered to be the same as the same species as the Enterobacterium (Eubacterium hallii) of the same species as Enterobacterium cloacae, clostridium ulcerans (Fusobacterium ulcerans), fusobacterium mutans (Fusobacterium varium), chicken of duck origin (Gallibacterium anatis), budding formate (Gemmiger formicilis), gordonia palustris (Gemmiger formicilis), hafnia alvei, bile helicobacter (Gemmiger formicilis), canadian helicobacter (Gemmiger formicilis), helicobacter canis (Gemmiger formicilis), homosamara helicobacter (Gemmiger formicilis), kiwi helicobacter (Gemmiger formicilis), pameti helicobacter (Gemmiger formicilis), young avian helicobacter (Gemmiger formicilis), helicobacter pylori (Gemmiger formicilis), rodent helicobacter (Gemmiger formicilis), gemmiger formicilis helicobacter (Gemmiger formicilis), mosaic (Herbazoliromassian), double-shaped hupermann (Gemmiger formicilis), filiform hupermann (Gemmiger formicilis), hupermann bacillus (Gemmiger formicilis), hupermann lactobacillus (Gemmiger formicilis), lactobacillus acidophilus (Gemmiger formicilis) and lactobacillus acidophilus (Gemmiger formicilis) of the like, lactobacillus casei (Lactobacillus buchneri), lactobacillus casei, lactobacillus curvatus (Lactobacillus curvatus), lactobacillus delbrueckii (Lactobacillus delbrueckii), lactobacillus fermentum (Lactobacillus fermentum), lactobacillus gasseri (Lactobacillus gasseri), lactobacillus helveticus (Lactobacillus helveticus), lactobacillus johnsonii, lactobacillus inertia (Lactobacillus iners), lactobacillus enterica (Lactobacillus intestinalis), lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus murinus (Lactobacillus murinus), lactobacillus paracasei (Lactobacillus paracasei), lactobacillus plantarum, lactobacillus reuteri (Lactobacillus reuteri), lactobacillus rhamnosus, lactobacillus rumen (Lactobacillus ruminis), lactobacillus sake (Lactobacillus sakei), lactobacillus salivarius (Lactobacillus salivarius), lactobacillus curvatus (Lactobacillus fermentum), lactobacillus erlenneatus (Lactobacillus ultunensis), lactobacillus vaginalis (Lactobacillus vaginalis), lactobacillus plantarum subspecies (Lactobacillus vaginalis) and Leuconostoc mesenteroides (Lactobacillus vaginalis), leuconostoc pseudocatenulatum (Lactobacillus vaginalis), listeria Listeria (Listeria), listeria (Listeria, mentha monocytogenes), lactobacillus johnsonii (Lactobacillus vaginalis), lactobacillus megaterium (Lactobacillus vaginalis) and Methanomyces (Lactobacillus vaginalis) and Methanomonas megans (Lactobacillus vaginalis), bacteria of the order Mollicutes (Mollicutes), rhizomonas enterica (Murimonas intestini), neisseria kii (Neisseria macacae), acrylonitrile acidophilus (Nitriliruptor alkaliphilus), bacillus Marseis (Oceanobacillus massiliensis), acidobacter villosa (Odoribacter laneus), acidobacter visceral (Odoribacter splanchnicus), acidobacter nasi (Ornithobacterium rhinotracheale), oxalic acid producing bacteria (Oxalobacter formigenes), paenibacillus balun (Paenibacillus barengoltzii), paenibacillus chitin (Paenibacillus chitinolyticus), paenibacillus lautus (Paenibacillus chitinolyticus), paenibacillus thuringiensis (Paenibacillus chitinolyticus), paenibacillus sainii (Paenibacillus chitinolyticus), sarcina pseudobulb (Paenibacillus chitinolyticus), paenias diradica (Paenibacillus chitinolyticus) the bacterial strain comprises the following components of a. Gordonii (Paenibacillus chitinolyticus), a. Johnsonii (Paenibacillus chitinolyticus), a. Faecalis (Paenibacillus chitinolyticus), a. Xylanophila (Paenibacillus chitinolyticus), a. Saphenous (Paenibacillus chitinolyticus), a Micromonospora, a Pediococcus acidilactici (Paenibacillus chitinolyticus), a clostridium difficile (Paenibacillus chitinolyticus), a Rake peptone (Paenibacillus chitinolyticus), an obese peptone (Paenibacillus chitinolyticus), a Seagal peptone (Paenibacillus chitinolyticus), a Di-Moonese peptone (Paenibacillus chitinolyticus), a Kara succinate (Paenibacillus chitinolyticus), a Porphyromonas incendii (Paenibacillus chitinolyticus), a Porphyromonas upper field (Paenibacillus chitinolyticus), prevotella papyrifera (Prevotella baroniae), prevotella bipivaria, prevotella faecalis (Prevotella copri), prevotella denticola (Prevotella dentalis), prevotella iridis (Prevotella micans), prevotella polyose (Prevotella multisaccharivorax), prevotella stomatalis (Prevotella oralis), prevotella salivaria (Prevotella salivae), prevotella faecalis (Prevotella stercorea), prevotella true (Prevotella veroralis), propionibacterium acnes, propionibacterium greedingii (Propionibacterium avidum), propionibacterium freudenreichii, microbacterium lymphophilicum (Propionimicrobium lymphophilum), proteus mirabilis (Proteus mirabilis), proteus penri ATCC providencia (Providencia alcalifaciens), providencia rette (Providencia rettgeri), providencia rette (Providencia rustigianii), providencia stuartii (Providencia stuartii), pseudolysis hirsutum (Huang Tongjun), pseudomonas aeruginosa (Huang Tongjun), pseudomonas pale yellow (Huang Tongjun), ralstonia pileri (Huang Tongjun), rhaponticum hyaline (Huang Tongjun), rhaponticum texas (Huang Tongjun), morganum pigeon (Huang Tongjun), morganum seashore (Huang Tongjun), ralstonia faecalis (Roseburia faecis), ralstonia enterica (Huang Tongjun), ralstonia inulin (Huang Tongjun), the composition comprises (a) a bicyclic ruminococcus (Ruminococcus bicirculans), a ruminococcus buchneri (Ruminococcus bromii), a smart ruminococcus (Ruminococcus callidus), a still-in-pal ruminococcus (Ruminococcus champanellensis), a fecal ruminococcus (Ruminococcus faecis), an active ruminococcus (Ruminococcus gnavus), a yoghurt ruminococcus (Ruminococcus lactaris), an oval ruminococcus (Ruminococcus obeum), a ruminococcus species, a chain ruminococcus (Ruminococcus torques), a sarcina (Sarcina ventriculi), a intestinal fecal uniconas (Sarcina ventriculi), a anaerobic saikoku (Sarcina ventriculi), a shigella sonnei, a pear-shaped neisseria (Sarcina ventriculi), a staphylococcus epidermidis, a slow staphylococcus (Sarcina ventriculi), a staphylococcus nepalensis (Sarcina ventriculi), a streptococcus agarici (Sarcina ventriculi), a streptococcus agalactiae (Sarcina ventriculi), a streptococcus sphaerobicus (Sarcina ventriculi), streptococcus equi (Sarcina ventriculi), streptococcus (Sarcina ventriculi) and infant (Sarcina ventriculi), streptococcus sarkovatus (Streptococcus merionis), streptococcus mitis (Streptococcus mitis), streptococcus mutans, streptococcus stomatus (Streptococcus oralis), streptococcus ovis (Streptococcus ovis), streptococcus parahaemolyticus (Streptococcus parasanguinis), streptococcus suis (Streptococcus plurextorum), streptococcus suis (Streptococcus porci), streptococcus pyogenes, streptococcus salivarius (Streptococcus salivarius), streptococcus suis (Streptococcus sobrinus), streptococcus thermophilus (Streptococcus thermophilus), streptococcus tolhaote (Streptococcus thermophilus), streptomyces albus (Streptococcus thermophilus), micrococcus anamorphic (Streptococcus thermophilus), pseudomonas shi (Streptococcus thermophilus), sarcandid (Streptococcus thermophilus), wotztec (Streptococcus thermophilus), ethylene glycol-producing agrobacterium (Streptococcus thermophilus), streptococcus thermophilus bacillus (Streptococcus thermophilus), organic matter deep sea bacillus (Streptococcus thermophilus), streptococcus saikochiae (Streptococcus thermophilus), unknown species (unowns sp.), unknown species, campylobacter buchnsonii (vabacillus), atypical (Streptococcus thermophilus), mivir (Streptococcus thermophilus), different species (Streptococcus thermophilus), and vibrio vulgare (Streptococcus thermophilus) and vibrio need of the same.

In other embodiments, the targeted bacterial cells are those common in the skin microbiota, and are, without limitation, acetobacter faveolatum (Acetobacter farinalis), acetobacter malatus (Acetobacter malorum), acetobacter alfa (Acetobacter orleanensis), acetobacter fruit (Acetobacter sicerae), acetobacter anxiety (Achromobacter anxifer), acetobacter denitrificans (Achromobacter denitrificans), acetobacter sphaeroides Ma Depu (Achromobacter marplatensis), acetobacter febrile (Achromobacter spanius), acetobacter xylosoxidans subsp (Achromobacter xylosoxidans subsp. Xylosoxidans), konjak acidovorax (Acidovorax konjaci), acetophaga (Acidovorax radicis), acetobacter johnsonii (Acinetobacter johnsonii), actinomyces lemanensis (Actinomadura citrea), cynanchumus pseudolaris (Actinomadura coerulea), actinobacillus fibrosus (Actinomadura fibrosa), actinobacillus palmatidus (Actinomadura fibrosa), actinobacillus sanguineensis (Actinomadura fibrosa), actinobacillus nitrodactylothermophilus (Actinomadura fibrosa), actinobacillus verruckerii (Actinomadura fibrosa), actinobacillus verrucosa (Actinomadura fibrosa), actinobacillus angusta (Actinomadura fibrosa), actinobacillus angustifolia (Actinomadura fibrosa) and actinobacillus angustifolia (Actinomadura fibrosa), pseudomonas putida (Aeromonas bestiarum), aeromonas bivalve (Aeromonas bivalvium), aeromonas anguillarum (Aeromonas encheleia), aeromonas spae (Aeromonas eucrenophila), aeromonas hydrophila subspecies hydrophila (Aeromonas hydrophila subsp. Hydrophila), aeromonas curiosa (Aeromonas piscicola), aeromonas boricola (Aeromonas popoffii), aeromonas lineans (Aeromonas rivuli), aeromonas salmonicida pectolylis (Aeromonas salmonicida subsp. Pecies), aeromonas salmonicida subspecies (Aeromonas salmonicida subsp. Smithia), micrococcus kapri sewer (Amaricoccus kaplicensis), verona sewer coccus (Amaricoccus veronensis), A He Ye amino bacilli (Aminobacter aganoensis), acidobacter sajohnsonii (Aminobacter aganoensis), sargassum (Aminobacter aganoensis), sinoaminobacilli (Aminobacter aganoensis), sinorhizobacter polymorphus (Aminobacter aganoensis), flavobacteria yunnanensis (Aminobacter aganoensis), bacillus nator (Aminobacter aganoensis), bacillus sp, bacillus (Aminobacter aganoensis) and Bacillus (Aminobacter aganoensis), brevibacterium flavum (Bradyrhizobium huanghuaihaiense), rhizobium japonicum (Bradyrhizobium japonicum), brevibacterium flavum (Brevundimonas aurantiaca), brevibacterium intermedia (Brevundimonas intermedia), burkholderia pinnatifida (Burkholderia aspalathi), burkholderia georginata (Burkholderia choica), burkholderia koidz (Burkholderia cordobensis), burkholderia guangdaliensis (Burkholderia diffusa), burkholderia gracilii (Burkholderia insulsa), burkholderia cervi (Burkholderia rhynchosiae), burkholderia terrestris (Burkholderia rhynchosiae), burkholderia georginata (Burkholderia rhynchosiae), bulleria ganii (Burkholderia rhynchosiae), sludge on the ground (Burkholderia rhynchosiae), carbon dioxide gum fibrous bacteria (Burkholderia rhynchosiae), bulletia papilis (Burkholderia rhynchosiae), myxobacteria (Burkholderia rhynchosiae), geobacillus glabra (Burkholderia rhynchosiae), goldali (Burkholderia rhynchosiae), nannocheir-gold (Burkholderia rhynchosiae), nannovi (Burkholderia rhynchosiae), corynebacterium parvulus (Burkholderia rhynchosiae), corynebacterium parvophilus (Burkholderia rhynchosiae), and Corynebacterium parvum (Burkholderia rhynchosiae), copper-killing bacteria (), copper-bulimium chondrium (), copper-wilt, short bacillus (), leech skin Dewok's bacteria (), riboflavin de wok's bacteria, rice beneficial bacteria (), alcaliphilia bacteria (), adhesive sword bacteria (), american sword bacteria (), enterococcus casselifaciens (), enterococcus pseudobird (), enterococcus vietnamensis (), enterococcus aromaticum (), colibacillus, rhodobacter salicina (), xanthobacter halibut (), xanthobacter arvensis (), xanthobacter Flavobacterium cold (), gluconobacter freundii (), gluconobacter thailand (), gordonia alkane (), celastomonas sea (), celastomonas axillari (), celastomonas southern (), celastomonas olive (), celastomonas mobilis (), celastomonas variabilis (), porphyra crudus subspecies (subsp. Putei), bursaphenous vein (), bursaphenous vein structure-forming a complex, pseudomonas tzeri (Herminiimonas fonticola), cytophaga intermedia (Hydrogenophaga intermedia), cytophaga pseudo Huang Qing (Hydrogenophaga pseudoflava), klebsiella oxytoca, leuconostoc (Kosakonia sacchari), lactobacillus delbrueckii subspecies bulgaricus (Lactobacillus delbrueckii subsp. Bulgarisus), lactobacillus helson (Lactobacillus modestisalitolerans), lactobacillus plantarum Azoffia subspecies (Lactobacillus modestisalitolerans), monomonas pseudoginseng (Lactobacillus modestisalitolerans), lactobacillus fragrans (Lactobacillus modestisalitolerans), leuconostoc mesenchanensis (Lactobacillus modestisalitolerans), micro5237 (Lentzea albida), california (Lactobacillus modestisalitolerans), leuconostoc fleshy (Lactobacillus modestisalitolerans), leuconostoc citrifolia (Lactobacillus modestisalitolerans), leuconostoc mesenteroides (Lactobacillus modestisalitolerans subspecies), leuconostoc mesenteroides (Lactobacillus modestisalitolerans), leucomonas stenotrophomonas (Lactobacillus modestisalitolerans), leucomonas pseudomonas (Lactobacillus modestisalitolerans), leuconostoc mesenteroides (Lactobacillus modestisalitolerans), leuconostoc (Lactobacillus modestisalitolerans) and Magjia (Lactobacillus modestisalitolerans) are described as being able to take place in the sea state of the Magzfeldes (Lactobacillus modestisalitolerans), methylobacterium columbus (Methylobacterium haplocladii), methylobacterium syringae (Methylobacterium haplocladii), methylobacterium verrucosum (Methylobacterium haplocladii), methylobacterium zakii (Methylobacterium haplocladii), microbacterium oxydanum (Methylobacterium haplocladii), micromonospora schneideriana (Methylobacterium haplocladii), micromonospora bronze (Methylobacterium haplocladii), micromonospora citrifolia (Methylobacterium haplocladii), micromonospora colkochiana (Methylobacterium haplocladii), micromonospora echinomatosis (Methylobacterium haplocladii), micromonospora halophila (Methylobacterium haplocladii), micromonospora parkeri (Methylobacterium haplocladii), micromonospora maritima (Methylobacterium haplocladii), micromonospora nigrum (Methylobacterium haplocladii), micromonospora rhodochrous (Methylobacterium haplocladii), micromonospora rhizogenes (Methylobacterium haplocladii), micromonospora cescens (Methylobacterium haplocladii), micromonospora prandium (Methylobacterium haplocladii), mycobacterium globosa (Methylobacterium haplocladii), mycobacterium guani forest subspecies (Methylobacterium haplocladii), mycobacterium aureovorans (Methylobacterium haplocladii), mycobacterium angustiforme (Methylobacterium haplocladii), mycobacterium fortunae (Methylobacterium haplocladii) and Mycobacterium fortunai (Methylobacterium haplocladii), mycobacterium megaterium (Mycobacterium obuense), mycobacterium exoticum (Mycobacterium peregrinum), mycobacterium sainformis (Mycobacterium saopaulense), mycobacterium septicemia (Mycobacterium saopaulense), mycobacterium siamensis (Mycobacterium saopaulense), mycobacterium smegmatis (Mycobacterium saopaulense), neisseria microflavonoides (Mycobacterium saopaulense), nocardia (Mycobacterium saopaulense), sphingosine (Mycobacterium saopaulense), flavosporium (Novosporium (Mycobacterium saopaulense), flavobacterium pseudogenitalium (Mycobacterium saopaulense), agrobacterium city (Mycobacterium saopaulense), paraqua Gracille (Mycobacterium saopaulense), paraqua paratuberculosis (Mycobacterium saopaulense) phenazine b.sub.Klebsiella (Mycobacterium saopaulense), immobilized b.sub.Klebsiella (Mycobacterium saopaulense), white rot fungus b.sub.Klebsiella (Mycobacterium saopaulense), b.sub.Klebsiella sojae (Mycobacterium saopaulense), b.parakochia (Mycobacterium saopaulense), d.glucose-phaga (Mycobacterium saopaulense), c.ginseng m.sub.m (Mycobacterium saopaulense), c.rhodosporum (Mycobacterium saopaulense), p.jejuni (Mycobacterium saopaulense), p.melanogenesis (Mycobacterium saopaulense), p.propinqua.acnes subspecies (Mycobacterium saopaulense subsp.elemongatum), c.vulgaris (Proteus vulgares), providencia stuartii, pseudoalteromonas agaricus (Pseudoalteromonas agarivorans), pseudoalteromonas atlantica (Pseudoalteromonas atlantica), pseudoalteromonas coral (Pseudoalteromonas paragorgicola), pseudopteromania ferox (Pseudomonas asplenii), pseudomonsa alfa at arizhou university (Pseudomonas asuensis), pseudomonsa alfa (Pseudomonas benzenivorans), pseudomonsa cannabinus (Pseudomonas cannabina), pseudomonsa albopictus (Pseudomonas cissicola), pseudomonsa icum (Pseudomonas congelans), pseudomonsa kangardii (Pseudomonas costantinii), pseudomonsa sanguinea (Pseudomonas ficuserectae), pseudomonsa virens (pseudomonsa virginiana), pseudomonsa herbaceous (Pseudomonas graminis), pseudomonsa jersensis (Pseudomonas jessenii) Pseudomonas korea (Pseudomonas koreensis), pseudomonas korea, pseudomonas kumquat (Pseudomonas kunmingensis), pseudomonas marginalis (Pseudomonas marginalis), pseudomonas fragi (Pseudomonas marginalis), pseudomonas pseudostellariae (Pseudomonas marginalis), pseudomonas fragi (Pseudomonas marginalis), pseudomonas pseudostellaria (Pseudomonas poae), pseudomonas pseudoalcaligenes (Pseudomonas marginalis), pseudomonas putida (Pseudomonas marginalis), pseudomonas Lepidii (Pseudomonas marginalis), pseudomonas rhizomae (Pseudomonas marginalis), pseudomonas selenopropionii (Pseudomonas marginalis), pseudomonas animalis (Pseudomonas marginalis), pseudomonas culpri (Pseudomonas marginalis), pseudomonas culprit (Pseudomonas marginalis), pseudomonas alfa (Pseudomonas marginalis), pseudomonas alaninetobacter (Pseudomonas marginalis), pseudomonas pseudoalcalii (Pseudomonas marginalis), pseudomonas aminoxide (Pseudomonas marginalis), autotrophic Pseudonocardia (), pseudonocardia alfa (), pseudonocardia yunnanensis (), pseudonocardia sanguinea (), pseudomonas pseudolaris (), pseudomonas indici (), pseudomonas homoandrostana (), geohydrophilic (), pseudonocardia pustulata North polar thermophilic thermophilgraphs, preparation of the preparation of psychrophilic bacillus, psychrophilic bacillus in fish house, psychrophilic bacillus in lung, ginsenoside converted to sandy loam bacillus Rheinheimia japonica (), rheimia dream, rheimia soil, rheimia down, rheimia Texas, rheimia tilapia, rheimia, rheimia Rheizobium (Rhizobium alami) Azibuergeria (), rhizobium (Rhizobium binae), rhizobium japonicum (Rhizobium etli), rhizobium japonicum (Rhizobium fabae), rhizobium freudenreichii (), rhizobium freundii, gao Lu rhizobia (Rhizobium gallicum), loess rhizobia (Rhizobium loessense), sophora rhizobia (Rhizobium sophoriradicis), taibai rhizobia (Rhizobium taibaishanense), valley rhizobia (Rhizobium vallis), cowpea rhizobia (Rhizobium vignagae), cowpea rhizobia, yang Ling rhizobia (Rhizobium yanglingense), red rhodococcus beijerinae (Rhodococcus baikonurensis), rhodococcus indicus (Rhodococcus enclensis), red rhodochrous celebrata (Rhodoferax saidenbachensis), rickettsia canadensis (Rickettsia canadensis), rickettsia nigra (Rickettsia heilongjiangensis), huo Enli g Rickettsia honei, ralstonia (Rickettsia raoultii), rose semi-photosynthetic bacteria (Roseateles aquatilis), rose semi-photosynthetic bacteria(s) aquatics Salamae enterica (Salmonella enterica subsp. Salamae), serratia fig (Serratia ficaria), serratia hepiali (Serratia mycotis), serratia hepiali (Serratia vespertilionis), shewanella tidal (Shewanella aestuarii), shewanella decolorationis (Shewanella decolorationis), sphingomonas pastoris (Shewanella decolorationis), chlorophenol (Shewanella decolorationis), copper-resistant Sphingomonas (Sphingomonas cuprimristiens), chlamys (Shewanella decolorationis), coal ash sphingosine (Shewanella decolorationis), indian sphingosine, japanese sphingosine (Shewanella decolorationis), copper-resistant Sphingomonas (Shewanella decolorationis), sphingosine-assimilating bacteria (Sphingobium lactosutens), sphingomonas uniisland (Sphingomonas dokdonensis), sphingomonas pseudohaemolyticus (Sphingomonas pseudosanguinis), chinesemetic bacteria (Sphingopyxis chilensis), sphingomonas freburgensis (Sphingopyxis fribergensis), sphingomonas gracilis (Sphingopyxis granuli), sphingomonas indicus (Sphingopyxis indica), sphingomonas wilfordii (Sphingopyxis witflariensis), staphylococcus aureus (Staphylococcus agnetis), staphylococcus aureus subsp. Sp. Aureus, staphylococcus epidermidis, human staphylococcus neomycin septicemia subsp. Staphylococcus aureus subsp. Novobiocinus, staphylococcus nepalensis, staphylococcus saprophyticus Staphylococcus aureus (Staphylococcus aureus subsp. Bots) staphylococcus pinus subspecies (Staphylococcus aureus subsp. Carnaticus), streptomyces nigrum (Staphylococcus aureus), streptomyces malachite (Staphylococcus aureus), streptomyces ascending (Staphylococcus aureus), streptomyces febrifugae (Staphylococcus aureus), streptomyces griseus (Staphylococcus aureus), streptomyces olive green (Staphylococcus aureus), streptomyces ginseng root (Staphylococcus aureus), streptomyces dark-red (Staphylococcus aureus), streptomyces pseudovenezuela (Staphylococcus aureus), streptomyces antimycin-producing (Staphylococcus aureus), deposit Staphylococcus aureus (Staphylococcus aureus), tsukamurensis microvolvulus (Staphylococcus aureus), bulimia guanfaciens (Staphylococcus aureus), fossa alcaligenes (Staphylococcus aureus), xanthomonas arborescens (Staphylococcus aureus), xanthomonas carpet (Xanthomonas axonopodis), xanthomonas cassava (Xanthomonas cassavae), xanthomonas cucurbitaceae (Xanthomonas cucurbitae), xanthomonas thistle (Xanthomonas cynarae), xanthomonas bullosa (Xanthomonas cynarae), xanthomonas strawberry (Xanthomonas cynarae), xanthomonas ganii (Xanthomonas cynarae), xanthomonas perforii (Xanthomonas cynarae), xanthomonas pisi pea (Xanthomonas pisi), xanthomonas gingivalis (Xanthomonas cynarae), xanthomonas avid (Xanthomonas cynarae), nucelia knosis (Yersinia nurmii), stenotrophomonas flacci (Xanthomonas cynarae), cystine-splitting bacteria (Xanthomonas cynarae), acinetobacter guangdong (Xanthomonas cynarae), acinetobacter parvulus (Xanthomonas cynarae), acinetobacter parvos (Xanthomonas cynarae) Acinetobacter radiodurans (Xanthomonas cynarae), acinetobacter terrestris (Xanthomonas cynarae), acinetobacter mutans (Xanthomonas cynarae), actinomycetes californica (Xanthomonas cynarae), actinomycetes odontoides (Xanthomonas cynarae), actinomycetes europaeus (Xanthomonas cynarae), actinomycetes gossypii (Xanthomonas cynarae), actinomycetes gracilii (Xanthomonas cynarae), actinomycetes johnsonii (Xanthomonas cynarae), actinomycetes mosaic (Xanthomonas cynarae), actinomycetes mechnii, actinomycetes endometrii, actinomycetes neotami nitrate-free subspecies (Xanthomonas cynarae), actinomycetes lykochia, actinomycetes stomatitis (actinomycetes oris), actinomycetes zurich (Xanthomonas cynarae), actinobacillus campylobacter (Actinotignum schaalii), actinobacillus cassii (Aerococcus christensenii), rhodococcus ureae (Aerococcus urinae), microorum flavum (Aeromicrobium flavum), microorum malasii (Sha Mana), microorum malasii (Aeromicrobium tamlense), aeromonas Sha Mana (Sha Mana), coagulobacterium acidophilus (Sha Mana), rhodobacter lahnsonii (Sha Mana), rhodobacter sphaeroides (Sha Mana), rhodobacter lazei (Sha Mana), alcaligenes olivaceus (Sha Mana), alcaligenes marinus (Sha Mana), rhodobacter sphaeroides, rhodobacter gray (Sha Mana), metal-resistant bacillus (Sha Mana), kapri sewer-collecting coccus, vinylna sewer coccus, anaerobacter (Sha Mana), anaerobacter, haemolyticus (Sha Mana), anaerobacter vaginalis (Sha Mana), haemophilus (Sha Mana), bacillus (Sha Mana), rhodobacter sphaericus (Sha Mana), rhodobacter sphaeroides (Sha Mana), stenotrophomonas (Sha Mana), rhodobacter sphaeroides (Sha Mana) and microorum (Sha Mana), acidithiobacillus (Atopobium parvulum), acidithiobacillus gingivalis (Atopobium rimae), acidithiobacillus vaginalis (Atopobium vaginae), acidovorax faciens (Aureimonas altamirensis), acidovorax faciens (Aureimonas rubiginis), aspergillus oryzae (Azospira oryzae), azospirillum oryzae (Azospirillum oryzae), bacillus circulans (Bacillus circulans), bacillus delreniensis, bacillus fastidious (Bacillus fastidiosus), bacillus stearothermophilus (Bacillus lehensis), bacillus marinus (Bacillus maceranisidiis), bacillus rhizosphere (Bacillus rhizosphaerae), vibrio stonecrop (Bacteriovorax stolpii), bacteroides coagulans (Bacteroides coagulans), bacteroides multocida, bacteroides fragilis, bacteroides simplex, bacteroides vulgaris, bdellovibrio exocarpi (Bdellovibrio exovorus), planocarpus morganii (Belnapia moabensis), nap (Belnaa soli), butterbrio Hantaenii, butterbrio bunyasis (5268), brevibacterium candidum (5282), brevibacterium flavum (Brevibacterium iodinum) and Brevibacterium flavum (35), brevibacterium flavum (Brevundimonas albigilva), brevibacterium deficiency (Brevundimonas diminuta), brevibacterium vulgare (Brevundimonas vancanneytii), brevibacterium terrestris, geobacillus indicus (Calidifontibacter indicus), campylobacter conciseum, campylobacter cilexenatis (Campylobacter gracilis), campylobacter hominis (Campylobacter hominis), campylobacter rectus (Campylobacter rectus), campylobacter shaoxidas (Campylobacter showae), campylobacter ureae (Campylobacter ureolyticus), cellovorans, LIDefibrate carbon dioxide acidophilia (Capnocytophaga leadbetteri), cellophaga (Capnocytophaga ochracea), carbon dioxide phlegm forming fiber (Capnocytophaga sputigena), human heart bacillus (Capnocytophaga sputigena), valve heart bacillus (Capnocytophaga sputigena), clostridium clockii (Capnocytophaga sputigena), karakul's bacteria (Catonella mobili), huntii (Capnocytophaga sputigena), subsurface cavity bacteria (Capnocytophaga sputigena), dexose fiber (Capnocytophaga sputigena), vibrio vulgaris (Capnocytophaga sputigena), rhizoctonia solani (Capnocytophaga sputigena), fuscoporia (Capnocytophaga sputigena), talaromyces (Capnocytophaga sputigena), tacrolimus (Capnocytophaga sputigena) and Tacrolimus (Capnocytophaga sputigena) can be prepared, golden (Chryseomicrobium aureum), barballs baumannii (Cloacibacterium haliotis), noman's tube (Cloacibacterium normanense), noman's tube, klebsiella aerogenes, de-nitrifying Comamonas (Comamonas denitrificans), tu-juna (Comamonas terrigena), congest, non-fermented Corynebacterium lipo subspecies, aminogenes, mycobacterium aceti, myxogolden (Corynebacterium aurimucosum), myxogolden corynebacteria, coleus (Corynebacterium coeae), hard corynebacterium (Corynebacterium durum), freiburg (Corynebacterium freiburgense), grey corynebacterium (Corynebacterium glaucum), glycine corynebacterium (Corynebacterium glyciniphilum), rhizoctonia, corynebacterium mimicus (Corynebacterium imitans), katsuki's corynebacterium (Corynebacterium jeikeium), katsujie's corynebacterium, koroti (Corynebacterium kroppenstedtii), lipophilic corynebacterium (Corynebacterium lipophiloflavum), corynebacterium (Corynebacterium massiliense), mastitis corynebacterium (Corynebacterium mastitidis), corynebacterium maritimum (Corynebacterium), microbacterium, corynebacterium (35), corynebacterium parvulus (6365), leucophyllum (3565), leucophyllum (6365) and Leucophyllum (6365 Corynebacterium tuberculosis, corynebacterium tachyphaga (), corynebacterium mutans (), zoozoozoobacteria of the family blue subspecies (subsp. Caeruleus), copper-resistant bacteria, brevibacterium herbicolum (), antichlorethamonas excitothecium (), deinococcus hard, deinococcus antarcticus (), deinococcus mud (), deinococcus ficuum (), deinococcus geothermal, deinococcus radiodurans (), dermatophytes vaginalis (), dermatophytes (); western coccus (), desemzia incrta, rostellus, listeria turcica (), aerosol, lesion producing, dietzia orange (), dietzia catenulatum (), dietzia diptheriae (), dietzia vensis, leucococcus lazy, leucococcus rodent (), leuconostoc, milisesurus, emmerce, brevibacterium, saccharum sinensis (), cyrtymenia, enterobacter roseum () Enterococcus faecalis (Enterococcus aquimarinus), enterococcus faecalis, enterococcus olivaceus (Enterococcus olivae), erwinia rheum, eubacterium parvulus (Eubacterium eligens), eubacterium faciens (Eubacterium infirmum), eubacterium rectum, eubacterium crypticum (Eubacterium saphenum), eubacterium aceti (Eubacterium sulci), microbacterium mexicanum (Exiguobacterium mexicanum), fuscoporia fumigatus (Fei Kelan), rhodobacter salina-resistant, golgi apparatus, flavobacterium epidermidis (Fei Kelan), bacillus (Fei Kelan), flavobacterium flavum (Fei Kelan) resistant, flavobacterium perfringens (Fei Kelan), fuscoporia nucleatum subspecies polymorpha (Fei Kelan subsp. Polymorphum) the bacterial strain comprises a haemolytic twin coccus (Fei Kelan), a measles twin coccus (Fei Kelan), a palatal twin coccus (Fei Kelan), a haemophilus (Fei Kelan), a germ bacterium aquaticus (Fei Kelan), a germ bacterium (Fei Kelan) sludge, a gordonia aurea (Fei Kelan), a gordonia polymorpha (Fei Kelan), a gordonia polyisoprene (Fei Kelan), a streptococcus agatus (Fei Kelan), a haemophilus parainfluenza (Fei Kelan), a haemophilus phlegm (Fei Kelan), a sulfide mineralocorticoid bacterium (Fei Kelan), a smooth column bacterium orange (Fei Kelan), a hydrocarbon bacterium extensor (Fei Kelan), sea-borne bacteria (Idiomarina maris), anopheles bifidus (Janibacter anophelis), holozenia (Janibacter hoylei), america (Janibacter indicus), america (Janibacter limosus), america (Janibacter melonis), salicomia Herbacea (Jeotgalicoccus halophilus), human (Rong Kaijun), lasiosphaera (Rong Kaijun), de-nitrifying (Rong Kaijun), klebsiella stomatocace (Kingella oralis), klebsiella oxytoca (Rong Kaijun), nardostachyos (Rong Kaijun), pinus albolsteri (Rong Kaijun), kocuria atrinae, gastrokudo (Rong Kaijun), klebsiella keiki (Rong Kaijun), klebsiella multock (Rong Kaijun), tarukukuke (Rong Kaijun), analox (Rong Kaijun), alterna (Rong Kaijun), acidovorax (Rong Kaijun), lactobacillus crispa (Rong Kaijun), latifolia (Rong Kaijun), legionella (Rong Kaijun), latifer (Rong Kaijun), legionella (Rong Kaijun) and Trichinella (Rong Kaijun), wei De ciliated bacteria, garcinia lutescens (Luteimonas terricola), lysine-forming bacillus (Lysinibacillus fusiformis), lysobacter sojae (Lysobacter spongiicola), lysobacter spongiicola, rhodococcus lyceum (Lysobacter spongiicola), marsdenia tenacissima (Lysobacter spongiicola), alkali-resistant Marsdenia, golden Marsdenia (Massilia aurea), folding Marsdenia (Massilia pliata), emultocida, lasiosphaera (Lysobacter spongiicola), thermus sieboldii (Lysobacter spongiicola), methylobacillus santalinus (Lysobacter spongiicola), methylobacillus griseus (Lysobacter spongiicola), methylophilus griseus, methylobacillus griseus Bacillus thuringiensis (Lysobacter spongiicola), methylobacillus salty, methylobacillus oxalicum (Lysobacter spongiicola), methylobacillus syringiensis, methylobacillus sagittifolius, methylobacillus widely used (Lysobacter spongiicola), microbacterium calcoaceticus (Lysobacter spongiicola), microbacterium hydrothermalis (Lysobacter spongiicola), microbacterium hydrothermally, microbacterium lacticum (Lysobacter spongiicola), microbacterium lacticum, microbacterium left-producing (Lysobacter spongiicola), microbacterium palum (Lysobacter spongiicola), microbacterium petroleum (Lysobacter spongiicola), microbacterium roseum (Lysobacter spongiicola), microbacterium antarcticum (Lysobacter spongiicola), micrococcus yellow (Lysobacter spongiicola), micrococcus lizicus (Lysobacter spongiicola), micrococcus (Micrococcus terreus), microzyme orange (Microlunatus aurantiacus), microcosmic (Micropruina glycogenica), microbacterium angustifolium (Microvirga aerilata), microbacterium angustifolium, microbacterium underground, microbacterium cowpea (Microvirga vignae), microbacterium prandium, micrococcus aerobicus, clostridium timum (Mogibacterium timidum), moraxella bitarvensis (Moraxella atlantae), moraxella catarrhalis (Moraxella catarrhalis), moraxella moelleriensis (Morganella morganii subsp. Morganii), moraxella cold-resistant (Morganella psychrotolerans), mercurous merkohl (Murdochiella asaccharolytica), mycobacterium asiaticus (Mycobacterium asiaticum), mycobacterium midpart (Mycobacterium chubuense), mycobacterium saffron (Mycobacterium crocinum) Mycobacterium californicum (Mycobacterium gadium), mycobacterium holstein (Mycobacterium holsaticum), mycobacterium islanguis (Mycobacterium iranicum), mycobacterium renba (Mycobacterium longobardum), mycobacterium neogolden, mycobacterium megaterium, negative succinic acid-eating coccus (Negativicoccus succinicivorans), neisseria (Neisseria bacilliformis), neisseria stomatalis (Neisseria oralis), neisseria sicca (Neisseria sicca), neisseria micro-yellow, ai Huohu Neisseria (Nesterenkonia lacusekhoensis), rhizoctonia rhizosphere (Nesterenkonia rhizosphaerae), persepia guava (Nevskia persephonica), branch-like Var.valia (Nevskia ramosa), america salishanensis (Niabella yanshanensis), moisture Bai Ganjun (Niveibacterium umoris), nocardia danabaca (Nocarpia niwae), nocardia thailand, nocardia agaophila (Nocardioides agariphilus), nocardia diluta (Nocardioides dilutus), nocardia Jiang Huadao (Nocardioides ganghwensis), nocardia cistenii (Nocardioides hwasunensis), nocardioides hwasunensis, nocardia depositional (Nocardioides hwasunensis), coccocus ampullate (Nocardioides hwasunensis), neisseria malaensis (Nocardioides hwasunensis), sphingomonas rhodochrous (Nocardioides hwasunensis), sinorhizobium rhizosphere (Nocardioides hwasunensis), nostoc, hadoku (Nocardioides hwasunensis), paracoccus parahaemophilus (Nocardioides hwasunensis), paracoccus parahaemolyticus (Nocardioides hwasunensis), micrococcus parahaemolyticus (Nocardioides hwasunensis), nostochae parapsilosis (Nocardioides hwasunensis), nostochae (Nocardioides hwasunensis), nostoc, micromonad, egionella polyflagelliforme (Parviterribacter multiflagellatus), proteus gensengides, geobacillus aquaticus (Pedobacter aquatilis), geobacillus ginseng (Pedobacter ginsengisoli), geobacillus west (Pedobacter xixiisoli), peptococcus nigricans (Peptococcus niger), cookideas peptone (Peptoniphilus coxii), goba's peptone (Peptoniphilus gorbachii), rabbit peptone, klebsiella (Peptoniphilus koenoeneniae), lacrimago peptone (Peptoniphilus lacrimalis), streptococcus anaerobacter (Peptostreptococcus anaerobius), streptococcus koxidans (Peptostreptococcus stomatis), takava (Phascolarctobacterium faecium), xenophilus (Phenylobacterium haematophilum), phenylbacterium spinosa (Phenylobacterium kunshanense), klebsiella multocida (Pluralibacter gergoviae), polyporus polysaccharideus (Polymorphobacter multimanifer), porphyromonas mandshurica, porphyromonas gingivalis (Polymorphobacter multimanifer), porphyromonas mandson (Polymorphobacter multimanifer), purpusia vinosa (Polymorphobacter multimanifer), purpusia buchnsoni (Polymorphobacter multimanifer), purpa stenia steniae (Polymorphobacter multimanifer), purpureae (Polymorphobacter multimanifer) and Purpa valia stenia (Volva) of the human body, volva (Polymorphobacter multimanifer), prevotella (Prevotella histicola), prevotella intermedia, prevotella jejuni, prevotella small-spot (Prevotella maculosa), prevotella melanogenes, prevotella iridae (Prevotella multiformis), prevotella southizinensis (Prevotella nanceiensis), prevotella melanosis, prevotella stomatalis (Prevolella oris), prevotella gingivitis (Prevotella oulorum), prevotella pallidum (Prevotella pallens), prevotella pleurisy (Prevotella pleuritidis), prevotella saccharolytica (Prevotella saccharolytica), prevotella salivaria, prevotella Sha Shipu (Prevotella shahii), prevotella pedica (Prevotella timonensis) Prevotella in the vacuum chamber, propionibacterium acidophilum (Propionibacterium acidifaciens), propionibacterium acnes subspecies acnes (Propionibacterium acnes subsp. Acnes), propionibacterium acnes subspecies acnes, propionibacterium granulose (Propionibacterium granulosum), microzyme lymphophilicum, toxoplasma gondii (Propionispira arcuata), pseudomonas stutzeri (Pseudokineococcus lusitanus), pseudomonas aeruginosa, pseudomonas chenopodii (Pseudomonas chengduensis), pseudonocardia penoxsula (Pseudonocardia benzenivorans), pseudorhodozoon besseyi (Pseudorhodoplanes sinuspersici), psychrophilum haemophilus (Psychrobacter sanguinis), ginsenoside-converted sand loam, sea water Rheimeria (Rheinheimera aquimaris), rhizobium (Rhizobium alvei), large Tian Genliu, rhizobium rosei (Rhizobium larrymoorei), rhizobia (Rhizobium rhizoryzae), rhizobia (Rhizobium rhizoli), rhizobia taibaiensis, rhizobia cowpea, glycine Luo Nahe bacillus (Rhodanobacter glycinis), rhodobacter verrucosa (Rhodobacter veldkampii), rhodococcus indicum, rhodococcus beam (Rhodococcus fascians), rhodococcus beam, rhodochrous cycloaliphatic (Rhodovarius lipocyclicus), campylobacter ranolae (Rivicola pingtungensis), rhodobacter oxyvorans (Rothia nasimurium), rosenbergii (3835), rhodobacter aerogenes (Roseomonas aerilata), rhodomonas aquaticus (Roseomonas aquatica), rhodopseudomonas mucilaginosa (Roseomonas mucosa), rhodopseudomonas rosea (Roseomonas rosea), rhodomonas vinifera (Roseomonas vinacea), rhodobacter aerogenes (Rothia), rhodobacter guli (Rothia amara e), rhodococcus equi (Rothia dentocariosa), rhodochrous endophyte (Rothia endophytica), rhodobacter mucilaginosa (Rothia mucilaginosa), rhodochrous nasolae (Rothia nasimurium), rhodobacter oxydans (5245), rhodochrous (Saccharopolyspora rectivirgula), rhodobacter oxydans (5282), rhodochrous (Saccharopolyspora rectivirgula) and other than can be expressed in the absence of the human being, rhodobacter oxydans (Saccharopolyspora rectivirgula) and rhodochrous (Saccharopolyspora rectivirgula), stereomonas acter (Selenomonas artemidis), acidovorax faciens (Selenomonas infelix), acidovorax faciens (Selenomonas noxia), acidovorax phlegm (Selenomonas sputigena), shewanella tidis, schmidwiella satellite (Shuttleworthia satelles), mu Lexi Meng Sishi (Simonsiella muelleri), shewanella aerobiotic (Skermanella aerolata), anti-Shewanella stibium (Skermanella stibiiresistens), shewanella sparsifolia (Slakia exigua), peacock Dan Qiujun (Smaragdicoccus niigatensis), xuepennisia (Sneathia sanguinegens), rhodobacter soil (Solirubrobacter soli), sphingobacterium mud (Solirubrobacter soli), bacillus megaterium (Solirubrobacter soli), and Solirubrobacter soli Bacillus thuringiensis (Solirubrobacter soli), kyoto sphingobacterium (Solirubrobacter soli), sphingobi (Solirubrobacter soli), sphingobacterium polysomum (Solirubrobacter soli), sphingobi (Solirubrobacter soli), sphingomonas sp, sphingomonas (Solirubrobacter soli), sphingomonas gingivalis (Solirubrobacter soli), sphingomonas paradoxes (Solirubrobacter soli) and Sphingomonas sp (Solirubrobacter soli), sphingomonas formosensis, sphingomonas gei, korea Sphingomonas Sphingomonas hankookensis, korea Sphingomonas Sphingomonas koreensis, kyoto Sphingomonas Sphingomonas kyeonggiensis, sphingomonas brickkilver Sphingomonas laterariae, sphingomonas sp Sphingomonas mucosissima, sphingomonas oligophenol Sphingomonas oligophenolica, sphingomonas pseudoblood, cyhingomonas zizaniae Sphingomonas sediminicola, sphingomonas salicina Sphingomonas yantingensis, sphingomonas yunnanensis Sphingomonas yunnanensis, sphingomonas indicus, sphingomonas wetland spirochesis, spongilla mesophilic vinegar producing strain Sporacetigenium mesophilum, spongilla viscosa fibrous strain Sporocytophaga myxococcoides staphylococcus aureons (Staphylococcus auricularis), staphylococcus epidermidis, staphylococcus hominis, staphylococcus lugdunensis, staphylococcus petiolus, pseudomonas febrile (Staphylococcus pettenkoferi), pseudomonas putida (Stenotrophomonas koreensis), pseudomonas putida (Stenotrophomonas rhizophila), pseudomonas putida, streptococcus agalactiae, streptococcus canis (Streptococcus canis), streptococcus crisis (Streptococcus cristatus), streptococcus gordonae, streptococcus infantis, streptococcus intermedium (Streptococcus intermedius), streptococcus mutans, streptococcus oligofermentus (Streptococcus oligofermentans), streptococcus stomatus, streptococcus sanguineus (Streptococcus sanguinis), streptomyces griseus (Streptomyces iconiensis), streptomyces Yang Linlian (Streptomyces yanglinensis), streptococcus aquaticus (Tabrizicola aquatica), bacillus thuringiensis (Tahibacter caeni), stannum (Tannerella forsythia), thermomyces salvinus (Tepidicella xavieri), thermomyces lanuginosus (Tepidimonas fonticaldi), agrobacterium luteus (Terracoccus luteus), tetracoccus luteus (Tessaracoccus flavescens), thermus thermophilus (Thermus thermophilus), thermomyces lanuginosus, arthrobacter, arthrospira amylovorus (Treponema amylovorum), treponema pallidum, treponema medum (Treponema lecithinolyticum), thermomyces lanuginosus (Turicella otitidis) Xuezurich bacillus, oligocarbohydrate coryneform bacteria (Undibacterium oligocarboniphilum), shrimp water coryneform bacteria (Undibacterium squillarum), salmon roaming coccus (Vagococcus salmoninarum), campylobacter canmanii, vibrio merrill (Vibrio metschnikovii), yellow marigold bacteria (Xanthobacter tagetidis), aerobiotic xenobiotics, anti-arsenophilus (Xenophilus arseniciresistens), micrococcus aurantiacus (yimelia lutea), bai Jimo mann bacteria (Zimmermannella alba), zimerman bacteria (Zimmermannella bifida) and/or Zoogloea (Zoogloea caeni).

In other embodiments, the targeted bacterial cells are those commonly found in vaginal microbiota, and is not limited to, acinetobacter (Acinetobacter antiviralis), acinetobacter baumannii, acinetobacter calcoaceticus (Acinetobacter calcoaceticus), acinetobacter johnsonii, actinomyces marseis (Actinobaculum massiliense), actinomyces shasii (Actinobaculum schaalii), actinomyces European, actinomyces gracilis, actinomyces israei, actinomyces mairei, actinomyces naeslundii, actinomyces lyxodentis, actinomyces zurich, actinomyces genitourinary (Actinomyces urogenitalis), actinomyces viscosus (Actinomyces viscosus), pneumococcus koani, pneumococcus ureae, pneumococcus grass, aeromonas anguillarum, aeromonas salmonicida, phillips mosaic (Afipia massiliensis), agrobacterium tumefaciens (Agrobacterium tumefaciens) aquatic psychrophila (Algoriphagus aquatilis), vibrio oldine (Aliivibrio wodanis), eimeria fenhnsonii, pediococcus otoi (Alloiococcus otitis), prevotella tannanensis (Alloprevotella tannerae), isovesicular multi-vitamin A, epoxy-feeding alternating red bacillus (Altererythrobacter epoxidivorans), oxalate ammonia-philic bacteria (Ammoniphilus oxalaticus), emammothecium bei (Amnibacterium kyonggiense), hydrogen-producing anaerobic coccus, lactolytic anaerobic coccus, mo Daoke anaerobic coccus, obesity-related anaerobic coccus (Anaerococcus obesiensis), anaerobic coccus praecox, tetranectia anaerobic coccus (Anaerococcus tetradius), anaerobic coccus vaginalis, double anaerobic spherical bacteria (Anaeroglobus geminatus), procino anaerobic bacillus (Anoxybacillus pushchinoensis), micro water bacillus, seal steal bacillus (Arcanobacterium phocae), brevibacterium flavum (Arthrobacter aurescens), achromobacter oxydans, micromirabilis (Atopobium minutum), micromirabilis gingivalis, micromirabilis vaginalis, avian Bacillus (Avibacterium gallinarum), bacillus acidophilus (Bacillus acidicola), bacillus megaterium (Bacillus atrophaeus), bacillus cereus (Bacillus cibi), bacillus paradoxus (Bacillus coahuilensis), bacillus pumilus (Bacillus gaemokensis), bacillus methanolica (Bacillus methanolicus), bacillus australis (Bacillus oleronius), bacillus pumilus, bacillus arensis (Bacillus shackletonii), bacillus thermosporus (Bacillus sporothermodurans), bacillus subtilis, and Bacillus glaucomatous (Bacillus wakoensis), bacillus Wei Enshi, bacillus tebuergerianus (Bacillus weihenstephanensis), paenium (Bacteroides barnesiae), paenibacillus coagulans Bacillus subtilis, bacillus bifidus, bifidobacterium longum subspecies, bifidobacterium pseudocatenulatum, bacillus thuringiensis, bacillus faecalis, bacteroides simplex, bacteroides vulgaris, bacteroides xylan, bacteroides actylovorus (Bacteroides zoogleoformans), bacillus thuringiensis (Barnesiella viscericola), bacillus subtilis at the center of cell molecular biology (Bhargavaea cecembensis), bifidobacterium adolescentis, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium longum subsp.infafricantis, bifidobacterium pseudocatenulatum, bifidobacterium Shi Kadu, bifidobacterium volvuli, bifidobacterium volcanicum, bifidobacterium parvum, bruella hydrogenotrophic, bruella ovata, bruella producing, brevibacterium faecalis (Brachybacterium faecium), soybean slow rooting tumor bacteria, brevibacterium majus (Brevibacterium mcbrellneri), brevibacterium otoi (Brevibacterium otitidis), brevibacterium febrile, legionella slow (Bulleidia extructa), burkholderia fungus (Burkholderia fungorum), burkholderia phenolicus (Burkholderia phenoliruptix), cellulolytic bacteria (Caldicellulosiruptor saccharolyticus), caldimonas taiwanensis, campylobacter gracilis, campylobacter mansoni, campylobacter phlegm (Campylobacter sputorum), campylobacter urealyticus, carbon dioxide phaga, human heart bacillus, klebsiella, chlamydia trachomatis, chlamydophila abortus (Chlamydophila abortus), cartilage fungus (Chondromyces robustus) Flavobacterium aquaticum (Chryseobacterium aquaticum), citrobacter bergii, corynebacterium nomanii (Cloacibacterium normanense), clostridium kali (Clostridium cavendishii), clostridium dog (Clostridium colicanis), clostridium jisoyase (Clostridium jejuense), clostridium perfringens, clostridium multi-branch, clostridium soxhlet (Clostridium sordellii), clostridium viride (Clostridium viride), comamonas terrestris, corynebacterium crowded, corynebacterium appendicitis (Corynebacterium appendicis), corynebacterium colestis, corynebacterium polygluconate (Corynebacterium glucuronolyticum), corynebacterium glutamicum (Corynebacterium glutamicum), corynebacterium kaki, corynebacterium korotkohl, corynebacterium lipophilic yellow, corynebacterium parvum, corynebacterium mucicum, corynebacterium distiller's yeast (Corynebacterium nuruki), corynebacterium pseudogenital tract (Corynebacterium pseudogenitalium), corynebacterium pyruvate-producing, corynebacterium uniquely, corynebacterium striatum (Corynebacterium striatum), corynebacterium tuberculosis stearate, corynebacterium xerosis, leng Leng-philium (Cryobacterium psychrophilum), brevibacterium wilt, dermatobacterium acnes, dermatobacterium greedy (Cutibacterium avidum), cellulomophaga xylan (Cytophaga xylanolytica), deinococcus radiophilus (Deinococcus radiophilus), crane feather Tian Daier Ford (Delftia tsuruhatensis), vibrio desulphus (Desulfovibrio desulfuricans), listeria cloudiness Dai A, listeria aerobics, listeria invaginata Dai A listeria invaginated (Dialister pneumosintes), listeria propionicum Dai A, dicus chrysanthemi (Dickeya chrysanthemi), long-chain dormer, eglinium tarda Alternaria alternata (Eggerthia catenaformis), egger Ai Kenjun, aerobacter gasbag, enterobacter albophilum, enterobacter cloacae, enterococcus avium, enterococcus durans, enterococcus faecalis, enterococcus faecium (Enterococcus hirae), erwinia persicae (Erwinia persicina), eubacterium rheum officinale, erwinia toletaa, escherichia coli, fei Gesen Escherichia (Escherichia fergusonii), eubacterium breve (Eubacterium brachy), eubacterium picki, eubacterium entanglement (Eubacterium brachy), eubacterium rectum, eubacterium cryptii, eubacterium aceti, eubacterium eubacterium uligerum (Eubacterium brachy), microbacterium aceti (Eubacterium brachy), facklamia ignova, enterobacter praecox, leptogenic bacteria (Filifactor alocis), fusobacterium megaterium, fusobacterium just, fusobacterium nucleatum, fusobacterium denticola Zhou Suo, gardnerella vaginalis, gemela vanica (Gemella asaccharolytica), gemela bergii (Gemela bergeri), geomelanuginosa, xemophilia coccus, geobacillus stearothermophilus (Geobacillus stearothermophilus), geobacillus thermode (Geobacillus)

thermocatenulatus), the enzyme Geobacillus thermoglucosidase (Geobacillus)

thermoglucosidase), geobacillus (Geobacter grbiciae), streptococcus elegans, haemophilus dulcitus (Haemophilus ducreyi), haemophilus haemolyticus (Haemophilus haemolyticus), haemophilus parahaemolyticus (Haemophilus parahaemolyticus), haemophilus parahaemolyticus, hafnia alvei, halomonas farnesii (Halomonas phoceae), halomonas melitensis (Halomonas venusta), rhodospirillum septicum (Herbaspirillum seropedicae), purple blue bacillus (Janthinobacterium lividum), human Rong Kai, klebsiella granulosa (Klebsiella granulomatis), klebsiella oxytoca, klebsiella pneumoniae, lactobacillus acidophilus, lactobacillus amylovorus (Lactobacillus amylovorus), lactobacillus brevis, lactobacillus vaginalis (Lactobacillus coleohominis), lactobacillus crispatus, lactobacillus curvatus lactobacillus delbrueckii, lactobacillus fermentum, lactobacillus grignard, lactobacillus helveticus, lactobacillus inertia, lactobacillus jensenii (Lactobacillus jensenii), lactobacillus johnsonii, lactobacillus caligenes (Lactobacillus kalixensis), lactobacillus sanfranciscensis (Lactobacillus kefiranofaciens), lactobacillus sauerkraut (Lactobacillus kimchicus), lactobacillus northwest (Lactobacillus kitasatonis), lactobacillus mucosae (Lactobacillus mucosae), lactobacillus bread (Lactobacillus panis), lactobacillus paracasei, lactobacillus plantarum, lactobacillus bridge (Lactobacillus pontis), lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus salivarius, lactobacillus ernata, lactobacillus vaginalis, lactobacillus lactis, lactobacillus buchneri, leuconostoc mesenteroides, leuconostoc citreum, leuconostoc mesenteroides (Leuconostoc garlicum), leuconostoc lactis, leuconostoc mesenteroides, lysomonas (Lysinimonas kribbensis) of the Korean institute of life and engineering, indomonas indoletica (Mageeibacillus indolicus), marine bacillus (Maribacter orientalis), haemomonas mutans (Marinomonas protea), sea-island helicobacter (Marinosporium, acidovorax sessiliflorus, megasphaera elhardtii (Megasphaera elsdenii), megasphaera micronutans (Megasphaera micronuciformis), rhizobium feverdanum (Mesorhizobium amorphae) in amorpha, methylobacillus radiodurans (Methylobacterium radiotolerans), bacillus methylotrophicus (Methylotenera versatilis), microbacterium halophilum (Microbacterium halophilum), micrococcus luteus, microbacterium paradiseae (Microterricola viridarii), acinetobacter kefir (Mobiluncus curtisii), acidovorax shlii (Mobiluncus mulieris), and Bacillus thuringiensis Moraxella glycerinum (Moorella glycerini), moraxella mulina (Moraxella osloensis), morganella morganii, indolenin Mo Lishi producing bacteria (Moryella indoligenes), mercduo-gram-bacteria, mycoplasma enteroides (Mycoplasma alvi), mycoplasma genitalium (Mycoplasma genitalium), mycoplasma hominus (Mycoplasma hominis), mycoplasma mice (Mycoplasma variens), mycoplasma salivarius (Mycoplasma salivarium), neisseria succinogenes, neisseria flavum (Neisseria flava), neisseria gonorrhoeae, neisseria mucilaginosa (Neisseria muricata), neisseria flavum, branch-like Zernia, mirabilis soil (Nevskia soli), alkaline nitrile-decomposing bacteria, zosteronibacterium visceratis, oligotrophic bacillus urethrai (Oligella urethralis), rhizoctonia gingivalis, bacillus amyloliquefaciens (Paenibacillus amylolyticus), paenibacillus humicola (Paenibacillus humicus), paenibacillus feed (Paenibacillus pabuli), paedena bacillus (Paenibacillus pasadenensis), paederia pinnatifida (Paenibacillus pini), paederia tonic (Paenibacillus validus), pantoea agglomerans (Pantoea agglomerans), paralopecuroides faecalis, paraorta carnivora (Paraburkholderia caryophylli), paraoccus jejuni, streptomyces abscess (Parastreptomyces abscessus), micromonospora, bevereign Guan Guojiao (Pectobacterium betavasculorum), soft rot pectobacter carotovorans (Pectobacillus carotovorum), pediococcus lactis, pediococcus ethanogenus (Pediococcus ethanolidurans), geobacillus fluvialis (Pedobacter alluvionis), geobacillus alensis (Pedobacter wanjuense), pseudomonas aquaticus (Paenibacillus pini), pediococcus melanogaster, metropolium irradiae (Peptoniphilus asaccharolyticus), geobacillus gordonii, leptophaeophilus, leptophaga, inone peptone (Peptoniphilus indolicus), lacrophila, streptococcus (Peptoniphilus massiliensis), streptococcus anaerobusta (673), streptococcus (7975), propionibacterium cold light emitting (3575), porphyromonas rhodobacter sphaeromonas, porphyromonas p.p.p.p.p.p.m (3575), porphyromonas p.p.p.p.m, makino (35) and Propionii (35) of Porphyromonas sp.sp.sp.sp.sp.sp.45, P.sp.sp.sp.sp.sp.45), prevotella, prevotella spell (Prevotella bergensis), prevotella two, prevotella cheek (Prevotella buccae), prevotella cheek (Prevotella buccalis), prevotella coloris (Prevotella colorans), prevotella faecalis, prevotella humana, prevotella denticola, prevotella deglycosis (Prevotella disiens), prevotella intermedia, prevotella lozenii (Prevotella loescheii), levobacteria Ma Shipu, prevotella melanogenes, prevotella iridae, prevotella melanogenes, prevotella stomatitis, prevotella ruminalis (Prevotella ruminicola), prevotella Sha Shipu, prevotella faecalis Levobacter calcoaceticus, prevotella calcoaceticus, microbacterium lymphophilium, proteus mirabilis, pseudomonas pinophilum (Pseudomonas abietaniphila), pseudomonas aeruginosa, pseudomonas amygdalina (Pseudomonas amygdali), pseudomonas azotoformans (Pseudomonas azotoformans), pseudomonas aeruginosa (Pseudomonas chlororaphis), pseudomonas quagensis (Pseudomonas pseudostellaria, pseudomonas fluorescens (Pseudomonas fluorescens), pseudomonas flavescens (Pseudomonas fulva), pseudomonas pseudostellaria (Pseudomonas lutea), pseudomonas stutzeri, pseudomonas oleaginis (Pseudomonas oleovorans), pseudomonas orientalis (Pseudomonas orientalis), pseudomonas pseudoalcaligenes, pseudomonas psychrophila (Pseudomonas psychrophila), pseudomonas pseudomonana (Pseudomonas synxantha), pseudomonas syringae (Pseudomonas syringae), pseudomonas syringae (Pseudomonas fluorescens), pseudomonas tolaciens (Pseudomonas tolaasii), pseudomonas propionica (Pseudopropionibacterium propionicum), ralstonia aquatica (Rahnella aquatilis), ralstonia pisiformis, ralstonia solanacearum (Ralstonia solanacearum), ralstonia plantarum (Raoultella planticola), rhizobium carotovorum (Rhizobacter dauci), rhizobium stibium crudus, rhodococcus fasciculi, rhodopseudomonas palustris (Rhodopseudomonas palustris), ralstonia enterica, ralstonia inulin-producing, ralstonia mucilaginosa, ruminococcus brotica, ruminococcus livens, ruminococcus sprain, haemophilus kadi (Sanguibacter keddieii), bacillus salmon sediment (Sediminibacterium salmoneum), zylobacter bovis (Selenomonas bovis), serratia sanguinea (Serratia fonticola), serratia liquefaciens (Serratia liquefaciens) Serratia marcescens, shewanella alga (Shewanella algae), shewanella amazonensis (Shewanella amazonensis), shigella boydii (Shigella boydii), shigella sonnei, shigella sparsa, shewanella amniovora (Sneathia amanii), serratia, mo Erlei Bacillus (Solobacterium moorei) alone, cellophane (Sorangium cellulosum), albizia, sphingomonas japonica, sphingomonas cornfield (Sphingobium yanoikuyae), sphingomonas veronii (Sphingomonas wittichii), sarcina seawater (Sporosarcina aquimarina), staphylococcus aureus, staphylococcus aurei, staphylococcus cephali (Staphylococcus capitis), staphylococcus epidermidis, staphylococcus haemolyticus (Staphylococcus haemolyticus), human staphylococci (Staphylococcus hominis), ludune staphylococci, saprophytic staphylococci, staphylococcus schneider (Staphylococcus schleiferi), staphylococcus similis (Staphylococcus simiae), staphylococcus mimicus (Staphylococcus simulans), staphylococcus wovensis (Staphylococcus warneri), stenotrophomonas maltophilia, acetobacter (Stenoxybacter acetivorans), streptococcus agalactiae, angina streptococcus, streptococcus australis, streptococcus equi, streptococcus bigeminicus (streptococcus mutans), streptococcus infantis, streptococcus intermedium, streptococcus paris, streptococcus sea (Streptococcus marimammalium), streptococcus mitis, streptococcus mutans, streptococcus stomatis, streptococcus parahaemolyticus (Streptococcus phocae), streptococcus pneumoniae (Streptococcus pseudopneumoniae), streptococcus salivarius, streptococcus thermophilus, waltz Wo Sisa, fusartan, haemophilus (Terrahaemophilus aromaticivorans), dental Treponema, haemophilus (Treponema maltophilum), microcosm (Treponema parum), venturi (Treponema vincentii), streptococcus berkovickers (Trueperella bernardiae), streptococcus pensis (5286), streptococcus equi, bacillus (Viridibacillus arenosi), bacillus parvobacteria (Virgibacillus proomii), pseudomonas aeruginosa (Virgibacillus proomii), pseudomonas aeruginosa (Streptococcus pseudopneumoniae), streptococcus equi, pseudomonas aeruginosa (Virgibacillus proomii), streptococcus equi, pseudomonas aeruginosa (Streptococcus pseudopneumoniae), streptococcus equi, pseudomonas (Virgibacillus proomii), streptococcus equi, pseudomonas (Streptococcus pseudopneumoniae), streptococcus equi) and other bacteria (Streptococcus pseudopneumoniae Weissella food (Weissella cibaria), weissella soil (Weissella soli), xanthomonas campestris (Xanthomonas campestris), xanthomonas bullosa (Xanthomonas vesicatoria), phaeophyta Zhuo Beier (Zobellia laminariae), and/or Achromobacter cladi (Zoogloea ramigera).

In one embodiment, the targeting bacterium is Escherichia coli.

In one embodiment, the targeting bacteria are dermatophytes acnes, more particularly acne-related dermatophytes from the genetic lineage (phylogroup) IA1 or RT4, RT5, RT8, RT9, RT10 or Clone Complex (CC) CC1, CC3, CC4, more particularly ST1, ST3, ST 4.

Thus, the first type of bacteriophage disclosed herein, and thus the bacteriophage particles or phage-derived delivery particles of the present invention, may specifically target (e.g., specifically target) bacterial cells from any one or more of the above-described bacterial genera and/or species, particularly to specifically deliver a payload.

In one embodiment, the targeted bacteria are pathogenic bacteria. The targeting bacteria may be toxic bacteria.

The targeting bacteria may be antibacterial resistant bacteria, preferably selected from the group consisting of ultra-broad spectrum beta-lactamase (ESBL) producing escherichia coli, ESBL klebsiella pneumoniae, vancomycin Resistant Enterococci (VRE), methicillin Resistant Staphylococcus Aureus (MRSA), multi-drug resistant (MDR) acinetobacter baumannii, MDR enterobacter species, and combinations thereof. Preferably, the targeting bacteria may be selected from the group of E.coli strains producing an ultra-broad spectrum of beta-lactamase (ESBL).

Alternatively, the targeting bacteria may be bacteria of a microbiome of a given species, preferably bacteria of a human microbiota.

In certain embodiments, the targeted bacterial cells are from a species or strain different from the producer bacterial cells.

Hybrid helper phage system and hybrid helper phage

The invention also relates to a hybrid helper phage system comprising:

(i) At least one phage DNA packaging gene as defined in the "producer bacterial cells" section above derived from lytic bacteriophage as defined in the "bacteriophage and bacteriophage derived genes" section above,

(i') at least one phage structural gene as defined in the "producer bacterial cells" section above derived from said lytic bacteriophage,

(i ") optionally, at least one phage gene involved in phage regulation as defined in the" producer bacterial cells "section above, derived from said lytic bacteriophage, and

(ii) At least one gene derived from a non-lytic bacteriophage as defined in the "bacteriophage and bacteriophage-derived genes" section above, as defined in the "producer bacterial cells" section above, which is involved in phage excision/insertion, phage DNA replication and/or phage regulation,

Wherein the genes (i), (i') and (ii) are contained in separate nucleic acid molecules or separate nucleic acid molecules, and

wherein the hybrid helper phage system does not comprise any expressed phage structural genes derived from the non-lytic bacteriophage.

In the context of the present invention, the term "hybrid helper phage system" means a set of at least one nucleic acid molecule, preferably at least two separate nucleic acid molecules, comprising the genes (i), (i '), optionally (i') and (ii) defined above, which allow the production of lytic phage particles and/or lytic phage-derived delivery vehicles by a producer bacterial cell comprising the system, wherein the genes (i), (i '), optionally (i') and (ii) are distributed over the at least two separate nucleic acid molecules when the system comprises at least two separate nucleic acid molecules.

As used herein, the term "nucleic acid" refers to a sequence of at least two nucleotides covalently linked together, which may be single-stranded or double-stranded, or contain portions of both single-stranded and double-stranded sequences. The nucleic acids of the invention may be naturally occurring, recombinant or synthetic. The nucleic acid may be in the form of a circular sequence or a linear sequence or a combination of both. The nucleic acid may be DNA (both genomic or cDNA) or RNA or a combination of both. The nucleic acid may contain any combination of deoxyribonucleotides and ribonucleotides and any combination of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, 5-hydroxymethylcytosine, and isoguanine. Chem rev.2016oct 26, weigele et al; 116 Further examples of modified bases that can be used in the present invention are described in detail in (20) 12655-12687. The term "nucleic acid" also includes any nucleic acid analog that may contain other backbones including, without limitation, phosphoramides, phosphorothioates, phosphorodithioates, O-methylphosphite linkages and/or deoxyribonucleotides and ribonucleotide nucleic acids. The invention also includes any combination of the above features of the nucleic acids.

In a particular embodiment, the genes (i), (i '), optionally (i') and (ii) are comprised in a bacterial chromosome, in particular in a producer bacterial cell chromosome. In more particular embodiments, the genes (i), (i '), optionally (i "), and (ii) are contained in the same region in the bacterial chromosome, and in alternative embodiments, the genes (i), (i'), optionally (i"), and (ii) are contained in different regions in the bacterial chromosome.

In alternative embodiments, the genes (i), (i'), optionally (i "), and (ii) are contained in separate plasmids. In another particular embodiment, the genes (i), (i '), optionally (i') and (ii) are all comprised in the same plasmid.

In another particular embodiment, the genes (i), (i'), optionally (i "), and (ii) are each independently contained in a bacterial chromosome or plasmid.

In a more particular embodiment, the genes (i), (i '), optionally (i') and (ii) are comprised in a hybrid helper phage.

Thus, in a particular embodiment, the hybrid helper phage system consists of hybrid helper phages comprising:

(i) At least one phage DNA packaging gene as defined in the "producer bacterial cell" section above, at least one phage structural gene as defined in the "producer bacterial cell" section above, and optionally at least one phage gene involved in phage regulation as defined in the "producer bacterial cell" section above, which is derived from lytic bacterial phage as defined in the "phage and gene derived from bacterial phage" section above, and

(ii) At least one gene as defined in the section "producing bacterial cells" above which is derived from a non-lytic bacteriophage as defined in the section "bacteriophage and bacteriophage-derived genes" above, which is involved in phage excision/insertion, phage DNA replication and/or phage regulation,

wherein the hybrid helper phage does not comprise any phage structural genes derived from the non-lytic bacteriophage.

By "helper phage" is meant herein a engineered phage that provides all necessary gene products for particle formation when using a phagemid vector. Helper phages typically have defective origins of replication or packaging signals and are therefore inefficient in self-packaging.

By "hybrid helper phage" is meant herein a engineered helper phage consisting of elements derived from at least lytic and non-lytic bacteriophages.

In a particular embodiment, the hybrid helper phage of the present invention is integrated as a prophage in the genome of the producer bacterial cell.

Production method

The invention further relates to a method for producing lytic phage particles or a lytic phage-derived delivery vehicle comprising:

(a) Providing a producer bacterial cell of the invention, and

(b) Inducing expression of at least one of said at least one phage structural gene and said phage DNA packaging gene in said producer bacterial cell, and assembling a product expressed by said at least one phage structural gene and said at least one phage DNA packaging gene, thereby producing a lytic phage particle or a lytic phage-derived delivery vehicle.

The induction step (b) may be carried out by any technique well known to the skilled person. In particular, as will be appreciated by the skilled person, the induction step will depend on the specific induction mechanism controlling the expression of the at least one of the phage structural gene and phage DNA packaging gene in the producer bacterial cell.

More particularly, the skilled artisan will appreciate that when the induction mechanism comprises at least one gene involved in phage excision/insertion, phage DNA replication and/or phage regulation derived from a non-lytic bacteriophage, the induction step will depend on the sequence-derived bacteriophage. In general, the induction step may be thermal induction (for phage naturally triggered by the signal or engineered repressor such as λci), small molecule inducers (depending on phage), any signal triggering SOS response (e.g. addition of mitomycin), etc.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It must be noted that the singular forms "a", "an" and "the" as defined herein include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells (e.g., a population of such cells). Similarly, reference to "a nucleic acid" includes one or more of such nucleic acids.

Although the invention has been described in conjunction with specific embodiments, many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations as fall within the spirit and broad scope of the appended claims.

All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that chapter titles are used, they should not be interpreted as necessarily limiting.

The following examples are given for the purpose of illustrating various embodiments of the invention and are not intended to limit the invention in any way.

Sequence(s)

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Brief Description of Drawings

Fig. 1: lambda genome tissue (packaging variant). The structural operon and the anti-terminator protein Q that allows transcription of the late structural operon are marked with red lines. The figure was adapted from Rajagopala et al, BMC Microbiol 11,213 (2011)).

Fig. 2: titration of T7 phagemid generated in lambda-T7 preheterozygous phage system. The production strain CY-L7 contains a payload p1883. Left panel, titration of MG 1655; right panel, titration of KEIO-waaG.

Fig. 3: titration of T7 phagemid generated in lambda-T7 preheterozygous phage system. Left panel, titration of MG 1655. Right panel, titration of KEIO-waaG. From left to right: left) payload p1883+p1885 (rapidly degrading T7 RNA polymerase); in) payload p1883+p1884 (medium strength degraded T7 RNA polymerase); right) payload p1883 only.

Fig. 4: identification of propionibacterium freudenreichii phages by PCR. PCR of ORF3 and ORF5 was performed on all phage suspensions. BW4 from plaques 1-3 gives a band for both orf3 and orf5 at the expected size. The ladder was GeneRuler 1kb+.

Fig. 5: lysogenic Pf0s14253 repeat immunization of infection. Left diagram: upper agar of Pf0s2841 with 4 different BW-like phage suspension spots. Right figure: upper agar of Pf0s14253 with 4 different BW-like phage suspension spots.

Fig. 6: high induction of BW4 phage following mitomycin C treatment. Left diagram: upper agar with Pf0s2841 from culture supernatant spots without mitomycin C (MMC) -induced Pf0s14253 (ND: undiluted to dilution 10) ^-3 ). Right figure: upper agar with Pf0s2841 from culture supernatant spots with 0.5. Mu.g/ml mitomycin C-induced Pf0s14253 (ND: undiluted to dilution 10) ^-7 )。

Fig. 7: genomic organization of BW4 and PAC7 bacteriophages. BW4 and PAC7 genomic tissues are similar, and both putative structural operons (indicated by arrows) contain packaging, head, tail and lysis modules.

Fig. 8: construction of chimeric BW4-PAC7 prophages. The pAN514 suicide plasmid was transformed into strain Pf1s22499 containing BW4 prophage. Chloramphenicol-based selection was used to select double crossover between Left Homology Arm (LHA) and Right Homology Arm (RHA). The prophage obtained is a chimeric body containing the structural operon, first BW4 gp1, then gp1-gp14 of PAC7, and the remainder of the BW4 structural gene (gp 15-gp 25) after the chloramphenicol selection cassette (CmR).

Fig. 9: plasmid map of cosmid pAN 594.

Fig. 10: titration of PAC7 phage-derived particles. Left diagram: titration of Pf1s22904 plated on erythromycin. Right figure: titration of control suspension Pf1s22903 strain plated on erythromycin without carrying any cosmids.

Fig. 11: confirmation of streaked 8 colonies from phage-derived particle titration produced by Pf1s22904 by PCR. Upper graph: SLTS PCR (Scholz 2014) of streaked 8 colonies from phage-derived titration assays. The expected size is 612bp. The following figures: pAN 594-specific PCR of 8 colonies. The expected size is 769bp. The ladder is typically 1kb+.

Examples

Example 1:exchange of structural operons for lambda of lytic phage elements

The inventors believe that the phage may be regarded as a more or less large genetic circuit, the final output of which is to produce more phage particles. To this end, whether the phage is lytic, temperate, or chronic (e.g., a filamentous phage such as M13), the information encoded in its genome can be broadly classified according to the function it performs:

genes specific for insertion/excision (for temperate phages).

Genes specific for DNA replication, RNA transcription, etc. For example, some lytic phages encode their own RNA or DNA polymerase. Some genes modify the RNA polymerase of the host to be able to work beyond the terminator and if the prophage sequence is present in plasmid or linear plasmid form, some other genes are involved in the isolation of the prophage sequence.

Genes associated with defenses against host anti-phage mechanisms, degradation/modification of host elements to complete the cleavage cycle, super-rejection mechanisms or genes beneficial to the host.

Genes specific for DNA packaging: termination enzymes and accessory proteins, ligases, and the like.

-structural genes specific for the construction of protein capsids for DNA: in addition to stringent structural genes such as capsid genes, tape measures, fibers, substrates, etc., many other genes are required to assemble the components (chaperones, proteases) and proteins that can be packaged within the capsid, either as scaffolds or as leader proteins injected into the cell (e.g., RNA polymerase of phage N4 or some small leader proteins in other phages).

The last two categories (DNA packaging and structural genes) are deeply linked, as the packaging mechanism recognizes the pre-assembled header and DNA to be packaged, starting and stopping DNA packaging.

The inventors hypothesize that by extracting and differentiating all the modules defined above, it is in principle possible to construct systems containing all excision/insertion, replication and regulatory elements from one bacteriophage, in particular a non-lytic bacteriophage, and encoding the packaging/structural genes from another bacteriophage, in particular a lytic bacteriophage, since in principle they can be regarded as independent genetic modules.

In this example, it is referred to as the "structural element" of the protein required for DNA packaging and the structural protein required for assembly of mature viral particles.

Such "heterozygous structural phages" can be very advantageous for different methods, as:

species more suitable for laboratory work/mass production/safer may be used for the production of such particles, wherein the structural gene is from another species;

the regulatory elements of a well-characterized phage (e.g.lambda) driving different phage capsid production etc. can be used to construct pure phagemid-producing strains, and

and finally, a structural hybrid prophage (i.e. carried in the genome) can be constructed that drives the production of the lytic phage capsid.

This is the method developed herein. The coding system was used to generate a producer strain of pure lambda phagemid whose structural operon had been exchanged with the structural elements of the strictly lytic E.coli phage T7 (from small termination enzyme to STF gene, about 23 kb). A schematic diagram shows the organization of the lambda genome (figure 1).

In this system, the thermolabile version of prophage lambda contains all the regulatory elements required to ablate the prophage, duplicates the circularized genome and drives expression of long, late operons, including the presence of anti-terminator protein Q. When the plasmid containing the correct packaging signal (LTR of T7) is complemented, this will drive the assembly and packaging of the pure phagemid particles entirely based on other phages.

Construction of heterozygotes

The lambda prophage structural operon (SEQ ID NO: 7) was exchanged with the structural "operon" of lytic phage T7, from gp6.5 to gp19.5 (not an operon in the strict sense, since T7 RNA polymerase drives transcription of different mRNAs in this region), using the lambda red recombination system, starting from the production strain containing lambda prophages without cos sites (s 1965). Several further modifications were made:

removal of perforin and of the cleavage genes assumed in T7 (gp 17.5 and gp 18.5)

Recoding the 3' part of the gp19 DNA maturation protein and the intergenic region between gp19 and the next (i.e.gp 19.5) as explained below

All T7 RNA polymerase promoters remain intact, but no T7 RNA polymerase is added to the system.

The fully edited construct "operon" spans approximately 20kb (SEQ ID NO: 8). The final product was designated CY-L7 and was constructed without any specific recitation.

Production and titration

The payload that should be packaged by T7 was constructed, e.g., auser et al, RNA biol.2019apr;16 (4) 595-599, referred to as pJ23115-GFP T7 cos 2.0 (p 1883, SEQ ID NO: 9). The payload contains the 5' LTR required for T7 efficient packaging. The putative packaging region of this plasmid contains the 3' portion of gp19 and the intergenic region between gp19 and gp 19.5. For this reason, the 3' portion of gp19 is recoded prior to its insertion into the genome of the production strain, thus preventing recombination.

Next, the CY-L7 strain was transformed with the p1883 payload and produced as follows.

The overnight cultures were incubated in LB+5mM CaCl supplemented with chloramphenicol ₂ Diluted 1:6 and grown with shaking at 30℃for 30min. Thereafter, thermal shock was performed at 42℃for 45 minutes. Finally, the cultures were grown at 37℃with shaking for 3 hours. After this period of time, the cells were recovered by centrifugation and lysed using 3mL of B-PER protein extraction reagent, and the biological beads were removed by adding 600mg of detergent and incubated at room temperature with gentle shaking for 1 hour. Thereafter, the lysate was centrifuged at 10,000g for 10min and the supernatant was filtered through a 0.2 micron pore size membrane.

Lysates were titrated in E.coli MG1655 and KEIO-waaG, a derivative with a deletion of the waaG gene that has been shown to be necessary for T7 binding (Qiaron et al Proc Natl Acad Sci U S A. (2006) 103 (50): 19039-19044). If phagemids were produced, colonies should be detected only in the MG1655 strain, since KEIO-waaG does not contain the T7 receptor.

As shown in fig. 2, only a small number of colonies were detectable in the MG1655 column. This result is the first demonstration, strict lytic phage can be "tamed" and its structure and packaging genes by lysogenic (lambda) control, to produce a T7 based pure phagemid particle.

The titer obtained was very low, although a pure T7-based phagemid was produced. The inventors have attempted to increase titres by applying different rational methods. For example, it is known that transcription from a promoter within the 5' LTR is required for T7 plasmid or genomic packaging by T7 RNA polymerase (Chung et al, J Mol biol.1990Dec 20;216 (4): 927-38). In addition, the T7 genome was transcribed by its cognate RNA polymerase, and many different T7 promoters were found, even in regions encoding different structural elements (Dunn et al, J Mol biol.1983Jun5;166 (4): 477-535). This resulted in different mRNAs which were then treated by E.coli RNase III (Studier et al, "Processing of bacteriophage T RNAs by RNase III" Ed: thomas R.Russell, keith Brew, harvey Faber, julius Schultz, from Gene to Protein: information Transfer in Normal and Abnormal Cells, academic Press,1979, p.261-269). For both reasons, the production strain was trans-complemented with T7 RNA polymerase in an inducible plasmid under the control of the PhlF repressor.

Initially, transformation of the T7 RNA polymerase plasmid in the CY-L7 strain containing the p1883 payload did not produce colonies, presumably due to toxicity caused by leakage of the inducible pphlF promoter (data not shown). For this reason, two alternative plasmids encoding T7 RNA polymerase with two different degradation tags of different intensities were constructed (p 1884, SEQ ID NO:10; and p1885, SEQ ID NO: 11). The sequences of the T7 RNA polymerase encoded in these two plasmids are disclosed (SEQ ID NO:12 for version AAV and SEQ ID NO:13; SEQ ID NO:14 for version LVA and SEQ ID NO: 15). It has been demonstrated that by adding a degradation tag to a protein, the potential impact of leaky expression from a repressed promoter can be ameliorated (Fernandez-Rodriguez et al, nucleic Acids res. (2016)

44(13):6493-6502)。

Production was performed from strain CY-L7 carrying payload p1883 and supplemented with either plasmid p1884 or the T7RNA polymerase variant encoded in p1885, using the same protocol as described above. Lysates were then titrated against MG1655 or KEIO-waaG.

As shown in fig. 3, the introduction of T7RNA polymerase increased the obtained titre 100X (for medium degradation tags) or 1000X fold (for fast degradation tags) compared to the product carrying only the p1883 payload. The titer obtained in this system was about 2x 10 ⁶ TU/mL。

These experiments indicate that for certain types of phage, regulatory proteins, in this case T7RNA polymerase, which do not strictly fall within the structural classes defined above, may be required to improve or promote the packaging reaction, or to control the amount or processing of mRNA encoding the structural components.

Example 2:production of phage-derived particles of skin bacillus acnes.

Dermatophytes acne is one of the most common and abundant skin species (Kashaf et al, nat Microbiol 7,169-179 (2022)), where it colonises the sebaceous gland unit (PSU). Unlike the stratum corneum, the bacteria present in PSU are surrounded by living cells, especially keratinocytes, sebocytes and different immune cells (Kabashima et al, nat Rev Immunol 19,19-30 (2019)). Intimate contact between the dermatophytes acne and these cells may lead to beneficial or detrimental interactions. (BrUggemann et al, front Microbiol 12,673845 (2021)). The ability to genetically modify dermatophytes acnes before the applicant's new tools as disclosed in US applications US2022/135986 and US2022/135987 is significantly challenging. In these patent applications, the inventors describe for the first time the production of phage-derived particles of dermatophytes acnes using dermatophytes acnes as production strain.

In this example, the inventors used the Propionibacterium freudenreichii strain to produce Propionibacterium acnes phage-derived particles by exchanging structural genes from Propionibacterium freudenreichii prophage for structural genes of Propionibacterium acnes.

Results

Isolation of BW4 phages

Propionibacterium freudenreichii and related bacteriophages are known to be present in some dairy products (Gautier et al, (1995) Lait 75:427-434; gautier et al, (1995) appl. Environ. Microbiol.61:2572-2576; cheng et al, (2018) BMC Microbiology 18:19). Thus, the inventors screened cheese samples for the presence of propionibacteria phage or propionibacterium freudenreichii lysogens.

Different types of cheese samples were ground, resuspended in Reinforced Clostridium Medium (RCM) and incubated for 2 days at 30 ℃ under anaerobic conditions. After incubation, dilution of the culture was carried out in lithium glycerolate broth, a medium selective for propionibacteria (WO 1994017201), and at 30 ℃Incubate for 6 days. Final dilution in rcm+mitomycin C was incubated at 30 ℃ for 1 day to induce potential prophages. The induced cultures (0.2 μm) were filtered and labeled based on the different indicator strains. One of the samples resulted in the formation of a cloudy plaque on the upper agar of the propionibacterium freudenreichii strain Pf0s 2841. 3 individual plaques were isolated by two consecutive picks and streaks on Pf0s2841 and amplified on the upper agar of Pf0s 2841. For three different plaques, amplification resulted in phage suspension-10 ¹⁰ PFU/mL。

Two clusters of temperate dsDNA Propionibacterium freudenreichii phages (BW and BV) have been previously identified (Cheng et al, (2018) BMC Microbiology 18:19). Using PCR based on BW genome design from doucete phage (KX 620751), two different fragments were extracted:

ORF3 with AD1334 (SEQ ID NO: 16)/AD 1335 (SEQ ID NO: 17)

ORF5 with AD1336 (SEQ ID NO: 18)/AD 1337 (SEQ ID NO: 19).

The inventors can classify isolated phages as BW-like (fig. 4). Sequencing of ORF5 shows that all phages are likely identical and therefore from the same BW-like phage named BW 4.

Isolation of Pf0s2841 lysogens carrying BW4 phages

The inventors then isolated propionibacterium freudenreichii lysogens that carried BW4 phage as prophages. To this end, BW4 phage suspensions were spotted on strain Pf0s2841 and incubated for 3 days. The cloudy plaques were picked, resuspended and streaked. After 5 days, single colonies were obtained, several colonies were streaked and incubated a second and third time, and at each streak, after dnase treatment, the presence of the phage gene at the cohesive end (AD 1322 (SEQ ID NO: 20)/AD 1323 (SEQ ID NO: 21)) was checked by PCR to ensure the presence of phage but not phage particles.

After the third streaking, the colonies were grown as upper agar and the undiluted BW-like phage suspension was spotted on the putative lysogenic strain (Pf 0s 14253) and the ancestor strain (Pf 0s 2841). After incubation, clearance of BW13 and BW14 spots was observed for both strains, whereas clearance of Pf0s2841 was only observed in the case of BW4 spots (fig. 5). This indicates that strain Pf0s14253 was immune to re-infection with BW4 phage and carried BW4 prophages. The absence of immunity from BW14 and BW13 suggests that these phages may have different immune repressors.

BW4 prophage induction

To use BW4 lysogenic strains as production strains for phage-derived particles, the inventors first have to test the ability to produce high concentrations of BW4 phage upon induction of a lysis cycle. To this end, pf0s14253 was grown in the absence or presence of the antibiotic mitomycin C (MMC) known to induce prophages, and the presence of BW4 phage particles was titrated against the culture supernatant based on the indicator strain Pf0s 2841. A large number of BW4 phage particles, 7.4X10, were observed under mitomycin C-supplemented conditions (FIG. 6) ⁷ PFU/. Mu.L, 3.0X10 s relative to the conditions without mitomycin C ³ PFU/. Mu.L. This suggests that the BW4 prophage has a high dynamic range between lysis and lysogenic cycle under such conditions and demonstrates the potential of BW4 for the production of phage-derived particles.

Sequencing and annotation of BW4 phages

To engineer the BW4 phage design to produce the acnes skin phage-derived particles, the BW4 phage was sequenced. The BW4 phage suspension was subjected to DNA isolation (Promega Wizard DNA Clean-Up system) followed by Illumina sequencing. Raw reads were assembled into single contigs (contigs) using Spades and end-corrected by Sanger sequencing (SEQ ID NO: 22). Annotation was performed using a Phaster and manually selected based on homology to other BW-like phages (Cheng et al, (2018) BMC Microbiology 18:19).

BW-like phages have the typical genomic structure of other temperate phages, as described in Cheng et al, (2018) BMC Microbiology18:19, with one large putative structural operon (also called lytic operon) organized in different functional modules that are in transcription order: packaging, head, tail and lysis modules. Surprisingly, it was assumed that the first gene of the operon (gp 1) appears to be associated with HHpred-based DNA replication, as it contains domains similar to bifunctional primases and polymerase proteins. The other parts of the BW4 phage genome contain genes required for prophage integration/excision, DNA replication, DNA recombination, lytic/lysogenic cycle regulation, and other accessory proteins. This modular structure demonstrates the possibility of exchanging the genes required for BW4 phage capsid generation and phage genome packaging for their equivalents from the dermatophyte acne phage genome.

Isolation of dermatophyte acne PAC7 phage

The acnes were isolated from the skin of healthy volunteers. Simple application of the patch (Biore) to the nose allows extraction of the acne, which is resuspended in RCM, plated on MRS and incubated under anaerobic conditions at 37 ℃. For some of these plates, plaques were able to be observed on dense dermatophytes acnes. DPBS (dun phosphate buffered saline) was poured onto the plate and the potential phage resuspended and filtered to remove bacteria. The phage suspension was streaked onto plates and added to the upper agar of strain Ca0s 2345. Plates were incubated for 2 days and plaques were re-separated by three consecutive picks, streaks and top agar plates. Finally, the plaques were amplified on the upper agar using the Ca0s2345 strain and the resulting plaque suspension PEG precipitated. High titer was obtained when Ca0s 2345-based drop timing>10 ⁶ PFU/. Mu.L) of phage suspension.

Sequencing and annotation of PAC7 phage

The PAC7 phage suspension was subjected to DNA isolation (Promega Wizard DNA Clean-Up System) followed by Illumina sequencing. Raw reads were assembled into single contigs using Spades and end-corrected by Sanger sequencing (SEQ ID NO: 23). Annotation was performed using a Phaster and manually selected based on homology to other Propionibacterium acnes (Marinelli et al, (2012) mBio 3:e00279-12). Similar to the propionibacterium freudenreichii BW4 phage, a structural operon comprising modules for packaging, head-to-tail assembly and cell lysis was identified (fig. 7). HNH endonuclease has been identified as the last gene of phage (gp 45). Such endonucleases have been shown to be essential for efficient packaging (Quiles-Puchalt et al, (2014) Proc Nat. Acad. Sci. 111:6016-6021).

Construction of lysogenic Strain Using chimeric BW4-PAC7 prophages

The genes in the BW4 prophage structural operon, from small termination enzyme gp2 to included tape measure protein gp16, were replaced by the structural PAC7 genes from gp1 to gp14 (fig. 8). This was performed by homologous recombination using plasmid pAN514 (SEQ ID NO: 24), a P.freudenreichii suicide vector cloned in E.coli DH 10B. After transformation of the vector, a double crossover event was selected with propionibacterium freudenreichii (Pf 1s 22499) by chloramphenicol-based selection. The integrity of the chimeric BW4-PAC7 structural operon was globally confirmed by PCR and sanger sequencing of the entire chimeric structural operon.

Production and production of PAC7 derived particles from lysogenic strains harboring chimeric BW4-PAC7 prophages (production) Titration

To generate propionibacterium acnes phage-derived particles from propionibacterium freudenreichii BW4-PAC7 chimeric lysogens, pAN594 cosmid (fig. 9) containing PAC7 phage packaging signals (SEQ ID NO: 25), the five genes expressing PAC7 tail module (gp 15-gp 19) and the operon of gp45 endonuclease (SEQ ID NO: 26) as well as origins of replication that are functional in propionibacterium freudenreichii and dermatobacterium acnes (as disclosed in U.S. applications US2022/135986 and US 2022/135787) were transformed into Pf1s 22903. Transformants were streaked and grown in the presence of chloramphenicol (1. Mu.g/ml) to select for the presence of prophages and in the presence of erythromycin (2.5. Mu.g/ml) to select for pAN 594. At OD _600nm About 0.4, cultures were supplemented with 0.5. Mu.g/ml mitomycin C and grown overnight at 30℃under anaerobic conditions. After incubation, cells were collected by centrifugation, lysed by bead beating (2 x 20min with 0.1mm glass beads at 30 Hz), the supernatant filtered and the presence of phage-derived particles titrated on dermatophytes acnes Ca0s 2258.

Obtain every mu L toAt most 10 ² A potential transconductor (fig. 10). 8 colonies were streaked onto Brain Heart Infusion (BHI) erythromycin (5. Mu.g/mL) and confirmed as dermatophytes acnes and transconductors carrying pAN594 using PCR (FIG. 11).

Thus, the inventors have demonstrated for the first time that a phage derived particle of Propionibacterium acnes capable of delivering DNA into the skin bacillus acnes can be produced by exchanging the structural gene of the Propionibacterium freudenreichii with the structural gene of the phage of the skin bacillus acnes.

Materials and methods

Strains used and produced

Table 1:strains used and produced

Culture conditions

All incubations of the propionibacterium freudenreichii strain were under anaerobic conditions (Thermo Scientific ^TM Sachet Oxoid ^TM AnaeroGen) at 30 ℃.

All incubations of the dermatophytes acnes strains were carried out in an anaerobic chamber at 37 ℃.

Construction of Strain Pf1s22499

Deletion of packaging signals from BW4 prophages was performed by homologous recombination and CRISPR-Cas selection recombinants using the pAN241 propionibacterium vector cloned in e.coli and then transformed into the Pf0s14253 strain. The pAN241 vector contained a template for homologous recombination (SEQ ID NO: 27) and an FnCpf1 transcriptional cassette with crRNA targeting BW4 prophage cos.

Transformation protocol of propionibacterium freudenreichii

Transformation of propionibacterium freudenreichii was adapted from Brede, d.a. et al, appl Environ Microb, 8077-8084 (2005), substituting SLB (sodium lactate broth) medium for BHI.

Titration of phage-derived particles

Strain Ca0s2258 was streaked on BHI agar plates. Once dense growth on the plates was obtained, liquid cultures were established in BHI. After overnight incubation, the turbid cultures were concentrated 10X in BHI. Mu.l of cells were mixed with 10. Mu.l of diluted 1/10 and diluted 1/100 of the pure solution of phage-derived particles generated from Pf1s22904 or Pf1s22903 as negative controls. Samples were incubated at room temperature for 2h and then 1/10 serial dilutions were made in BHI, samples were incubated under anaerobic conditions at 37℃for 2h, after which 4. Mu.L spots were placed on BHI+5. Mu.g/mL erythromycin. Plates were incubated under anaerobic conditions at 37℃for 7 days.

Sequence listing

<110> Ai Lige biotechnology Co., ltd (ELIGO BIOSCIENCE)

<120> production of lytic phage

<130> EB2021-04b

<150> US63/187,531

<151> 2021-05-12

<150> US63/187,532

<151> 2021-05-12

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Met Lys Leu Ala Pro Asn Val Lys Gln Gln Ser Arg Gly Ile Lys His

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Ala Lys Gln Trp Gln Glu His Asp Ala Arg Met Ala Gly Asp Asn Glu

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Pro Pro Val Trp Leu Gly Glu Gln Gln Leu Ser Glu Leu Asp Lys Leu

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Gln Ile Val Pro Glu Gly Arg Lys Ser Val Arg Ile Phe Arg Ala Gly

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Tyr Leu Ala Pro Val Met Ile Lys Ala Ile Gly Gln Lys Leu Ala Ala

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Ala Gly Val Gln Asp Ala Asn Phe Tyr Pro Asp Gly Met His Gly Gln

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Lys Val Glu Asn Trp Arg Glu Tyr Leu Ala Arg Glu Arg Gln Asn Leu

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Ser Asp Gly Leu Val Ile Glu Leu Pro Val Lys Gln Lys Ala Gln Leu

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Ser Gln Met Ala Asp Ser Glu Arg Ala Gln Leu Leu Ala Asp Arg Phe

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Asp Gly Val Cys Val His Pro Glu Ser Glu Ile Val His Val Trp Cys

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Gly Gly Val Trp Cys Pro Val Ser Thr Met Glu Leu Ser Arg Glu Met

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Val Ala Ile Tyr Ser Glu His Arg Ala Thr Phe Ser Lys Arg Val Ile

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Asn Asn Ala Val Glu Ala Leu Lys Val Ile Ala Glu Pro Met Gly Glu

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Pro Ser Gly Asp Leu Leu Pro Phe Ala Asn Gly Ala Leu Asp Leu Lys

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Asn Gly Ile Glu Tyr Thr Pro Pro Ala Pro Gly Glu Asn Ile Arg Asp

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Asn Ala Pro Asn Phe His Lys Trp Leu Glu His Ala Ala Gly Lys Asp

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Pro Arg Lys Met Met Arg Ile Cys Ala Ala Leu Tyr Met Ile Met Ala

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Asn Arg Tyr Asp Trp Gln Met Phe Ile Glu Ala Thr Gly Asp Gly Gly

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Ser Gly Lys Ser Thr Phe Thr His Ile Ala Ser Leu Leu Ala Gly Lys

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Gln Asn Thr Val Ser Ala Glu Met Thr Ser Leu Asp Asp Ala Gly Gly

340 345 350

Arg Ala Gln Val Val Gly Ser Arg Leu Ile Val Leu Ala Asp Gln Pro

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Lys Tyr Thr Gly Glu Gly Thr Gly Ile Lys Lys Ile Thr Gly Gly Asp

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Pro Val Glu Ile Asn Pro Lys Tyr Glu Lys Arg Phe Thr Ala Val Ile

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Arg Ala Val Val Leu Ala Thr Asn Asn Asn Pro Met Ile Phe Thr Glu

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Arg Ala Gly Gly Val Ala Arg Arg Arg Val Ile Phe Arg Phe Asp Asn

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Ile Val Ser Glu Ala Glu Lys Asp Arg Glu Leu Pro Glu Lys Ile Ala

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Ala Glu Ile Pro Val Ile Ile Arg Arg Leu Leu Ala Asn Phe Ala Asp

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Ala Leu Ala Ile Lys Gln Gln Thr Asp Pro Val Ile Glu Phe Cys Gln

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gatgcaaatt tttaccctga tggtatgcac ggtcagaagg tggagaactg gcgcgaatat 360

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acgggggaat tttccccgca cacgccggag aactggatca ccacgcacaa cggcattgag 780

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cttgagcacg cagccggaaa agacccgcgc aagatgatgc gtatatgtgc cgcgctgtac 900

atgattatgg cgaaccggta cgactggcag atgtttattg aggccaccgg agacggcggg 960

agcggtaaaa gtacattcac acacatagcc agccttctgg cagggaaaca aaacacggta 1020

agcgctgaaa tgacatcgct tgatgatgct ggtgggcgtg cgcaggttgt cgggagtcgt 1080

cttatcgtcc tggcagacca gccgaaatat acaggcgaag gaacgggcat caagaaaatc 1140

acgggcggcg accccgtgga aattaacccg aaatatgaaa agcgttttac ggcggtaatc 1200

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tatgactctg ccgccgtcat aaaatggtat gccgaaaggg atgctgaaat tgagaacgaa 180

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atcgcaggtg aaattgccag tattctcgac gggctccccc tgtcggtgca gcggcgtttt 420

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gggtgtttat gcctacttta tagagcataa gcagcgcaac acccttatct ggttgccgac 840

ggatggtgat gccgagaact ttatgaaaac ccacgttgag ccgactattc gtgatattcc 900

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gaagcgtttc actaatgggc gtggcttctg gtgcctgggc ggtaaagcgg caaaaaacta 1020

ccgtgaaaag tcggtggatg tggcgggtta tgatgaactt gctgcttttg atgatgatat 1080

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aaagtccatc cgtggctcca cgccaaaagt gagaggcacc tgtcagattg agcgtgcagc 1200

cagtgaatcc ccgcatttta tgcgttttca tgttgcctgc ccgcattgcg gggaggagca 1260

gtatcttaaa tttggcgaca aagagacgcc gtttggcctc aaatggacgc cggatgaccc 1320

ctccagcgtg ttttatctct gcgagcataa tgcctgcgtc atccgccagc aggagctgga 1380

ctttactgat gcccgttata tctgcgaaaa gaccgggatc tggacccgtg atggcattct 1440

ctggttttcg tcatccggtg aagagattga gccacctgac agtgtgacct ttcacatctg 1500

gacagcgtac agcccgttca ccacctgggt gcagattgtc aaagactgga tgaaaacgaa 1560

aggggatacg ggaaaacgta aaaccttcgt aaacaccacg ctcggtgaga cgtgggaggc 1620

gaaaattggc gaacgtccgg atgctgaagt gatggcagag cggaaagagc attattcagc 1680

gcccgttcct gaccgtgtgg cttacctgac cgccggtatc gactcccagc tggaccgcta 1740

cgaaatgcgc gtatggggat gggggccggg tgaggaaagc tggctgattg accggcagat 1800

tattatgggc cgccacgacg atgaacagac gctgctgcgt gtggatgagg ccatcaataa 1860

aacctatacc cgccggaatg gtgcagaaat gtcgatatcc cgtatctgct gggatactgg 1920

cgggattgac ccgaccattg tgtatgaacg ctcgaaaaaa catgggctgt tccgggtgat 1980

ccccattaaa ggggcatccg tctacggaaa gccggtggcc agcatgccac gtaagcgaaa 2040

caaaaacggg gtttacctta ccgaaatcgg tacggatacc gcgaaagagc agatttataa 2100

ccgcttcaca ctgacgccgg aaggggatga accgcttccc ggtgccgttc acttcccgaa 2160

taacccggat atttttgatc tgaccgaagc gcagcagctg actgctgaag agcaggtcga 2220

aaaatgggtg gatggcagga aaaaaatact gtgggacagc aaaaagcgac gcaatgaggc 2280

actcgactgc ttcgtttatg cgctggcggc gctgcgcatc agtatttccc gctggcagct 2340

ggatctcagt gcgctgctgg cgagcctgca ggaagaggat ggtgcagcaa ccaacaagaa 2400

aacactggca gattacgccc gtgccttatc cggagaggat gaatgacgcg acaggaagaa 2460

cttgccgctg cccgtgcggc actgcatgac ctgatgacag gtaaacgggt ggcaacagta 2520

cagaaagacg gacgaagggt ggagtttacg gccacttccg tgtctgacct gaaaaaatat 2580

attgcagagc tggaagtgca gaccggcatg acacagcgac gcaggggacc tgcaggattt 2640

tatgtatgaa aacgcccacc attcccaccc ttctggggcc ggacggcatg acatcgctgc 2700

gcgaatatgc cggttatcac ggcggtggca gcggatttgg agggcagttg cggtcgtgga 2760

acccaccgag tgaaagtgtg gatgcagccc tgttgcccaa ctttacccgt ggcaatgccc 2820

gcgcagacga tctggtacgc aataacggct atgccgccaa cgccatccag ctgcatcagg 2880

atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc tatctgggca 2940

tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg aaagagtttg 3000

ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc atgatgattc 3060

gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc acctgggata 3120

ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag cgcatcagca 3180

acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt aatgacagcg 3240

gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg ccgcagaaat 3300

ggacatggat accccgtgag ttacccggcg ggcgcgcctc gttcattcac gtttttgaac 3360

ccgtggagga cgggcagact cgcggtgcaa atgtgtttta cagcgtgatg gagcagatga 3420

agatgctcga cacgctgcag aacacgcagc tgcagagcgc cattgtgaag gcgatgtatg 3480

ccgccaccat tgagagtgag ctggatacgc agtcagcgat ggattttatt ctgggcgcga 3540

acagtcagga gcagcgggaa aggctgaccg gctggattgg tgaaattgcc gcgtattacg 3600

ccgcagcgcc ggtccggctg ggaggcgcaa aagtaccgca cctgatgccg ggtgactcac 3660

tgaacctgca gacggctcag gatacggata acggctactc cgtgtttgag cagtcactgc 3720

tgcggtatat cgctgccggg ctgggtgtct cgtatgagca gctttcccgg aattacgccc 3780

agatgagcta ctccacggca cgggccagtg cgaacgagtc gtgggcgtac tttatggggc 3840

ggcgaaaatt cgtcgcatcc cgtcaggcga gccagatgtt tctgtgctgg ctggaagagg 3900

ccatcgttcg ccgcgtggtg acgttacctt caaaagcgcg cttcagtttt caggaagccc 3960

gcagtgcctg ggggaactgc gactggatag gctccggtcg tatggccatc gatggtctga 4020

aagaagttca ggaagcggtg atgctgatag aagccggact gagtacctac gagaaagagt 4080

gcgcaaaacg cggtgacgac tatcaggaaa tttttgccca gcaggtccgt gaaacgatgg 4140

agcgccgtgc agccggtctt aaaccgcccg cctgggcggc tgcagcattt gaatccgggc 4200

tgcgacaatc aacagaggag gagaagagtg acagcagagc tgcgtaatct cccgcatatt 4260

gccagcatgg cctttaatga gccgctgatg cttgaacccg cctatgcgcg ggttttcttt 4320

tgtgcgcttg caggccagct tgggatcagc agcctgacgg atgcggtgtc cggcgacagc 4380

ctgactgccc aggaggcact cgcgacgctg gcattatccg gtgatgatga cggaccacga 4440

caggcccgca gttatcaggt catgaacggc atcgccgtgc tgccggtgtc cggcacgctg 4500

gtcagccgga cgcgggcgct gcagccgtac tcggggatga ccggttacaa cggcattatc 4560

gcccgtctgc aacaggctgc cagcgatccg atggtggacg gcattctgct cgatatggac 4620

acgcccggcg ggatggtggc gggggcattt gactgcgctg acatcatcgc ccgtgtgcgt 4680

gacataaaac cggtatgggc gcttgccaac gacatgaact gcagtgcagg tcagttgctt 4740

gccagtgccg cctcccggcg tctggtcacg cagaccgccc ggacaggctc catcggcgtc 4800

atgatggctc acagtaatta cggtgctgcg ctggagaaac agggtgtgga aatcacgctg 4860

atttacagcg gcagccataa ggtggatggc aacccctaca gccatcttcc ggatgacgtc 4920

cgggagacac tgcagtcccg gatggacgca acccgccaga tgtttgcgca gaaggtgtcg 4980

gcatataccg gcctgtccgt gcaggttgtg ctggataccg aggctgcagt gtacagcggt 5040

caggaggcca ttgatgccgg actggctgat gaacttgtta acagcaccga tgcgatcacc 5100

gtcatgcgtg atgcactgga tgcacgtaaa tcccgtctct caggagggcg aatgaccaaa 5160

gagactcaat caacaactgt ttcagccact gcttcgcagg ctgacgttac tgacgtggtg 5220

ccagcgacgg agggcgagaa cgccagcgcg gcgcagccgg acgtgaacgc gcagatcacc 5280

gcagcggttg cggcagaaaa cagccgcatt atggggatcc tcaactgtga ggaggctcac 5340

ggacgcgaag aacaggcacg cgtgctggca gaaacccccg gtatgaccgt gaaaacggcc 5400

cgccgcattc tggccgcagc accacagagt gcacaggcgc gcagtgacac tgcgctggat 5460

cgtctgatgc agggggcacc ggcaccgctg gctgcaggta acccggcatc tgatgccgtt 5520

aacgatttgc tgaacacacc agtgtaaggg atgtttatga cgagcaaaga aacctttacc 5580

cattaccagc cgcagggcaa cagtgacccg gctcataccg caaccgcgcc cggcggattg 5640

agtgcgaaag cgcctgcaat gaccccgctg atgctggaca cctccagccg taagctggtt 5700

gcgtgggatg gcaccaccga cggtgctgcc gttggcattc ttgcggttgc tgctgaccag 5760

accagcacca cgctgacgtt ctacaagtcc ggcacgttcc gttatgagga tgtgctctgg 5820

ccggaggctg ccagcgacga gacgaaaaaa cggaccgcgt ttgccggaac ggcaatcagc 5880

atcgtttaac tttacccttc atcactaaag gccgcctgtg cggctttttt tacgggattt 5940

ttttatgtcg atgtacacaa ccgcccaact gctggcggca aatgagcaga aatttaagtt 6000

tgatccgctg tttctgcgtc tctttttccg tgagagctat cccttcacca cggagaaagt 6060

ctatctctca caaattccgg gactggtaaa catggcgctg tacgtttcgc cgattgtttc 6120

cggtgaggtt atccgttccc gtggcggctc cacctctgaa tttacgccgg gatatgtcaa 6180

gccgaagcat gaagtgaatc cgcagatgac cctgcgtcgc ctgccggatg aagatccgca 6240

gaatctggcg gacccggctt accgccgccg tcgcatcatc atgcagaaca tgcgtgacga 6300

agagctggcc attgctcagg tcgaagagat gcaggcagtt tctgccgtgc ttaagggcaa 6360

atacaccatg accggtgaag ccttcgatcc ggttgaggtg gatatgggcc gcagtgagga 6420

gaataacatc acgcagtccg gcggcacgga gtggagcaag cgtgacaagt ccacgtatga 6480

cccgaccgac gatatcgaag cctacgcgct gaacgccagc ggtgtggtga atatcatcgt 6540

gttcgatccg aaaggctggg cgctgttccg ttccttcaaa gccgtcaagg agaagctgga 6600

tacccgtcgt ggctctaatt ccgagctgga gacagcggtg aaagacctgg gcaaagcggt 6660

gtcctataag gggatgtatg gcgatgtggc catcgtcgtg tattccggac agtacgtgga 6720

aaacggcgtc aaaaagaact tcctgccgga caacacgatg gtgctgggga acactcaggc 6780

acgcggtctg cgcacctatg gctgcattca ggatgcggac gcacagcgcg aaggcattaa 6840

cgcctctgcc cgttacccga aaaactgggt gaccaccggc gatccggcgc gtgagttcac 6900

catgattcag tcagcaccgc tgatgctgct ggctgaccct gatgagttcg tgtccgtaca 6960

actggcgtaa tcatggccct tcggggccat tgtttctctg tggaggagtc catgacgaaa 7020

gatgaactga ttgcccgtct ccgctcgctg ggtgaacaac tgaaccgtga tgtcagcctg 7080

acggggacga aagaagaact ggcgctccgt gtggcagagc tgaaagagga gcttgatgac 7140

acggatgaaa ctgccggtca ggacacccct ctcagccggg aaaatgtgct gaccggacat 7200

gaaaatgagg tgggatcagc gcagccggat accgtgattc tggatacgtc tgaactggtc 7260

acggtcgtgg cactggtgaa gctgcatact gatgcacttc acgccacgcg ggatgaacct 7320

gtggcatttg tgctgccggg aacggcgttt cgtgtctctg ccggtgtggc agccgaaatg 7380

acagagcgcg gcctggccag aatgcaataa cgggaggcgc tgtggctgat ttcgataacc 7440

tgttcgatgc tgccattgcc cgcgccgatg aaacgatacg cgggtacatg ggaacgtcag 7500

ccaccattac atccggtgag cagtcaggtg cggtgatacg tggtgttttt gatgaccctg 7560

aaaatatcag ctatgccgga cagggcgtgc gcgttgaagg ctccagcccg tccctgtttg 7620

tccggactga tgaggtgcgg cagctgcggc gtggagacac gctgaccatc ggtgaggaaa 7680

atttctgggt agatcgggtt tcgccggatg atggcggaag ttgtcatctc tggcttggac 7740

ggggcgtacc gcctgccgtt aaccgtcgcc gctgaaaggg ggatgtatgg ccataaaagg 7800

tcttgagcag gccgttgaaa acctcagccg tatcagcaaa acggcggtgc ctggtgccgc 7860

cgcaatggcc attaaccgcg ttgcttcatc cgcgatatcg cagtcggcgt cacaggttgc 7920

ccgtgagaca aaggtacgcc ggaaactggt aaaggaaagg gccaggctga aaagggccac 7980

ggtcaaaaat ccgcaggcca gaatcaaagt taaccggggg gatttgcccg taatcaagct 8040

gggtaatgcg cgggttgtcc tttcgcgccg caggcgtcgt aaaaaggggc agcgttcatc 8100

cctgaaaggt ggcggcagcg tgcttgtggt gggtaaccgt cgtattcccg gcgcgtttat 8160

tcagcaactg aaaaatggcc ggtggcatgt catgcagcgt gtggctggga aaaaccgtta 8220

ccccattgat gtggtgaaaa tcccgatggc ggtgccgctg accacggcgt ttaaacaaaa 8280

tattgagcgg atacggcgtg aacgtcttcc gaaagagctg ggctatgcgc tgcagcatca 8340

actgaggatg gtaataaagc gatgaaacat actgaactcc gtgcagccgt actggatgca 8400

ctggagaagc atgacaccgg ggcgacgttt tttgatggtc gccccgctgt ttttgatgag 8460

gcggattttc cggcagttgc cgtttatctc accggcgctg aatacacggg cgaagagctg 8520

gacagcgata cctggcaggc ggagctgcat atcgaagttt tcctgcctgc tcaggtgccg 8580

gattcagagc tggatgcgtg gatggagtcc cggatttatc cggtgatgag cgatatcccg 8640

gcactgtcag atttgatcac cagtatggtg gccagcggct atgactaccg gcgcgacgat 8700

gatgcgggct tgtggagttc agccgatctg acttatgtca ttacctatga aatgtgagga 8760

cgctatgcct gtaccaaatc ctacaatgcc ggtgaaaggt gccgggacca ccctgtgggt 8820

ttataagggg agcggtgacc cttacgcgaa tccgctttca gacgttgact ggtcgcgtct 8880

ggcaaaagtt aaagacctga cgcccggcga actgaccgct gagtcctatg acgacagcta 8940

tctcgatgat gaagatgcag actggactgc gaccgggcag gggcagaaat ctgccggaga 9000

taccagcttc acgctggcgt ggatgcccgg agagcagggg cagcaggcgc tgctggcgtg 9060

gtttaatgaa ggcgataccc gtgcctataa aatccgcttc ccgaacggca cggtcgatgt 9120

gttccgtggc tgggtcagca gtatcggtaa ggcggtgacg gcgaaggaag tgatcacccg 9180

cacggtgaaa gtcaccaatg tgggacgtcc gtcgatggca gaagatcgca gcacggtaac 9240

agcggcaacc ggcatgaccg tgacgcctgc cagcacctcg gtggtgaaag ggcagagcac 9300

cacgctgacc gtggccttcc agccggaggg cgtaaccgac aagagctttc gtgcggtgtc 9360

tgcggataaa acaaaagcca ccgtgtcggt cagtggtatg accatcaccg tgaacggcgt 9420

tgctgcaggc aaggtcaaca ttccggttgt atccggtaat ggtgagtttg ctgcggttgc 9480

agaaattacc gtcaccgcca gttaatccgg agagtcagcg atgttcctga aaaccgaatc 9540

atttgaacat aacggtgtga ccgtcacgct ttctgaactg tcagccctgc agcgcattga 9600

gcatctcgcc ctgatgaaac ggcaggcaga acaggcggag tcagacagca accggaagtt 9660

tactgtggaa gacgccatca gaaccggcgc gtttctggtg gcgatgtccc tgtggcataa 9720

ccatccgcag aagacgcaga tgccgtccat gaatgaagcc gttaaacaga ttgagcagga 9780

agtgcttacc acctggccca cggaggcaat ttctcatgct gaaaacgtgg tgtaccggct 9840

gtctggtatg tatgagtttg tggtgaataa tgcccctgaa cagacagagg acgccgggcc 9900

cgcagagcct gtttctgcgg gaaagtgttc gacggtgagc tgagttttgc cctgaaactg 9960

gcgcgtgaga tggggcgacc cgactggcgt gccatgcttg ccgggatgtc atccacggag 10020

tatgccgact ggcaccgctt ttacagtacc cattattttc atgatgttct gctggatatg 10080

cacttttccg ggctgacgta caccgtgctc agcctgtttt tcagcgatcc ggatatgcat 10140

ccgctggatt tcagtctgct gaaccggcgc gaggctgacg aagagcctga agatgatgtg 10200

ctgatgcaga aagcggcagg gcttgccgga ggtgtccgct ttggcccgga cgggaatgaa 10260

gttatccccg cttccccgga tgtggcggac atgacggagg atgacgtaat gctgatgaca 10320

gtatcagaag ggatcgcagg aggagtccgg tatggctgaa ccggtaggcg atctggtcgt 10380

tgatttgagt ctggatgcgg ccagatttga cgagcagatg gccagagtca ggcgtcattt 10440

ttctggtacg gaaagtgatg cgaaaaaaac agcggcagtc gttgaacagt cgctgagccg 10500

acaggcgctg gctgcacaga aagcggggat ttccgtcggg cagtataaag ccgccatgcg 10560

tatgctgcct gcacagttca ccgacgtggc cacgcagctt gcaggcgggc aaagtccgtg 10620

gctgatcctg ctgcaacagg gggggcaggt gaaggactcc ttcggcggga tgatccccat 10680

gttcaggggg cttgccggtg cgatcaccct gccgatggtg ggggccacct cgctggcggt 10740

ggcgaccggt gcgctggcgt atgcctggta tcagggcaac tcaaccctgt ccgatttcaa 10800

caaaacgctg gtcctttccg gcaatcaggc gggactgacg gcagatcgta tgctggtcct 10860

gtccagagcc gggcaggcgg cagggctgac gtttaaccag accagcgagt cactcagcgc 10920

actggttaag gcgggggtaa gcggtgaggc tcagattgcg tccatcagcc agagtgtggc 10980

gcgtttctcc tctgcatccg gcgtggaggt ggacaaggtc gctgaagcct tcgggaagct 11040

gaccacagac ccgacgtcgg ggctgacggc gatggctcgc cagttccata acgtgtcggc 11100

ggagcagatt gcgtatgttg ctcagttgca gcgttccggc gatgaagccg gggcattgca 11160

ggcggcgaac gaggccgcaa cgaaagggtt tgatgaccag acccgccgcc tgaaagagaa 11220

catgggcacg ctggagacct gggcagacag gactgcgcgg gcattcaaat ccatgtggga 11280

tgcggtgctg gatattggtc gtcctgatac cgcgcaggag atgctgatta aggcagaggc 11340

tgcgtataag aaagcagacg acatctggaa tctgcgcaag gatgattatt ttgttaacga 11400

tgaagcgcgg gcgcgttact gggatgatcg tgaaaaggcc cgtcttgcgc ttgaagccgc 11460

ccgaaagaag gctgagcagc agactcaaca ggacaaaaat gcgcagcagc agagcgatac 11520

cgaagcgtca cggctgaaat ataccgaaga ggcgcagaag gcttacgaac ggctgcagac 11580

gccgctggag aaatataccg cccgtcagga agaactgaac aaggcactga aagacgggaa 11640

aatcctgcag gcggattaca acacgctgat ggcggcggcg aaaaaggatt atgaagcgac 11700

gctgaaaaag ccgaaacagt ccagcgtgaa ggtgtctgcg ggcgatcgtc aggaagacag 11760

tgctcatgct gccctgctga cgcttcaggc agaactccgg acgctggaga agcatgccgg 11820

agcaaatgag aaaatcagcc agcagcgccg ggatttgtgg aaggcggaga gtcagttcgc 11880

ggtactggag gaggcggcgc aacgtcgcca gctgtctgca caggagaaat ccctgctggc 11940

gcataaagat gagacgctgg agtacaaacg ccagctggct gcacttggcg acaaggttac 12000

gtatcaggag cgcctgaacg cgctggcgca gcaggcggat aaattcgcac agcagcaacg 12060

ggcaaaacgg gccgccattg atgcgaaaag ccgggggctg actgaccggc aggcagaacg 12120

ggaagccacg gaacagcgcc tgaaggaaca gtatggcgat aatccgctgg cgctgaataa 12180

cgtcatgtca gagcagaaaa agacctgggc ggctgaagac cagcttcgcg ggaactggat 12240

ggcaggcctg aagtccggct ggagtgagtg ggaagagagc gccacggaca gtatgtcgca 12300

ggtaaaaagt gcagccacgc agacctttga tggtattgca cagaatatgg cggcgatgct 12360

gaccggcagt gagcagaact ggcgcagctt cacccgttcc gtgctgtcca tgatgacaga 12420

aattctgctt aagcaggcaa tggtggggat tgtcgggagt atcggcagcg ccattggcgg 12480

ggctgttggt ggcggcgcat ccgcgtcagg cggtacagcc attcaggccg ctgcggcgaa 12540

attccatttt gcaaccggag gatttacggg aaccggcggc aaatatgagc cagcggggat 12600

tgttcaccgt ggtgagtttg tcttcacgaa ggaggcaacc agccggattg gcgtggggaa 12660

tctttaccgg ctgatgcgcg gctatgccac cggcggttat gtcggtacac cgggcagcat 12720

ggcagacagc cggtcgcagg cgtccgggac gtttgagcag aataaccatg tggtgattaa 12780

caacgacggc acgaacgggc agataggtcc ggctgctctg aaggcggtgt atgacatggc 12840

ccgcaagggt gcccgtgatg aaattcagac acagatgcgt gatggtggcc tgttctccgg 12900

aggtggacga tgaagacctt ccgctggaaa gtgaaacccg gtatggatgt ggcttcggtc 12960

ccttctgtaa gaaaggtgcg ctttggtgat ggctattctc agcgagcgcc tgccgggctg 13020

aatgccaacc tgaaaacgta cagcgtgacg ctttctgtcc cccgtgagga ggccacggta 13080

ctggagtcgt ttctggaaga gcacgggggc tggaaatcct ttctgtggac gccgccttat 13140

gagtggcggc agataaaggt gacctgcgca aaatggtcgt cgcgggtcag tatgctgcgt 13200

gttgagttca gcgcagagtt tgaacaggtg gtgaactgat gcaggatatc cggcaggaaa 13260

cactgaatga atgcacccgt gcggagcagt cggccagcgt ggtgctctgg gaaatcgacc 13320

tgacagaggt cggtggagaa cgttattttt tctgtaatga gcagaacgaa aaaggtgagc 13380

cggtcacctg gcaggggcga cagtatcagc cgtatcccat tcaggggagc ggttttgaac 13440

tgaatggcaa aggcaccagt acgcgcccca cgctgacggt ttctaacctg tacggtatgg 13500

tcaccgggat ggcggaagat atgcagagtc tggtcggcgg aacggtggtc cggcgtaagg 13560

tttacgcccg ttttctggat gcggtgaact tcgtcaacgg aaacagttac gccgatccgg 13620

agcaggaggt gatcagccgc tggcgcattg agcagtgcag cgaactgagc gcggtgagtg 13680

cctcctttgt actgtccacg ccgacggaaa cggatggcgc tgtttttccg ggacgtatca 13740

tgctggccaa cacctgcacc tggacctatc gcggtgacga gtgcggttat agcggtccgg 13800

ctgtcgcgga tgaatatgac cagccaacgt ccgatatcac gaaggataaa tgcagcaaat 13860

gcctgagcgg ttgtaagttc cgcaataacg tcggcaactt tggcggcttc ctttccatta 13920

acaaactttc gcagtaaatc ccatgacaca gacagaatca gcgattctgg cgcacgcccg 13980

gcgatgtgcg ccagcggagt cgtgcggctt cgtggtaagc acgccggagg gggaaagata 14040

tttcccctgc gtgaatatct ccggtgagcc ggaggcgtat ttccgtatgt cgccggaaga 14100

ctggctgcag gcagaaatgc agggtgagat tgtggcgctg gtccacagcc accccggtgg 14160

tctgccctgg ctgagtgagg ccgaccggcg gctgcaggtg cagagtgatt tgccgtggtg 14220

gctggtctgc cgggggacga ttcataagtt ccgctgtgtg ccgcatctca ccgggcggcg 14280

ctttgagcac ggtgtgacgg actgttacac actgttccgg gatgcttatc atctggcggg 14340

gattgagatg ccggactttc atcgtgagga tgactggtgg cgtaacggcc agaatctcta 14400

tctggataat ctggaggcga cggggctgta tcaggtgccg ttgtcagcgg cacagccggg 14460

cgatgtgctg ctgtgctgtt ttggttcatc agtgccgaat cacgccgcaa tttactgcgg 14520

cgacggcgag ctgctgcacc atattcctga acaactgagc aaacgagaga ggtacaccga 14580

caaatggcag cgacgcacac actccctctg gcgtcaccgg gcatggcgcg catctgcctt 14640

tacggggatt tacaacgatt tggtcgccgc atcgaccttc gtgtgaaaac gggggctgaa 14700

gccatccggg cactggccac acagctcccg gcgtttcgtc agaaactgag cgacggctgg 14760

tatcaggtac ggattgccgg gcgggacgtc agcacgtccg ggttaacggc gcagttacat 14820

gagactctgc ctgatggcgc tgtaattcat attgttccca gagtcgccgg ggccaagtca 14880

ggtggcgtat tccagattgt cctgggggct gccgccattg ccggatcatt ctttaccgcc 14940

ggagccaccc ttgcagcatg gggggcagcc attggggccg gtggtatgac cggcatcctg 15000

ttttctctcg gtgccagtat ggtgctcggt ggtgtggcgc agatgctggc accgaaagcc 15060

agaactcccc gtatacagac aacggataac ggtaagcaga acacctattt ctcctcactg 15120

gataacatgg ttgcccaggg caatgttctg cctgttctgt acggggaaat gcgcgtgggg 15180

tcacgcgtgg tttctcagga gatcagcacg gcagacgaag gggacggtgg tcaggttgtg 15240

gtgattggtc gctgatgcaa aatgttttat gtgaaaccgc ctgcgggcgg ttttgtcatt 15300

tatggagcgt gaggaatggg taaaggaagc agtaaggggc ataccccgcg cgaagcgaag 15360

gacaacctga agtccacgca gttgctgagt gtgatcgatg ccatcagcga agggccgatt 15420

gaaggtccgg tggatggctt aaaaagcgtg ctgctgaaca gtacgccggt gctggacact 15480

gaggggaata ccaacatatc cggtgtcacg gtggtgttcc gggctggtga gcaggagcag 15540

actccgccgg agggatttga atcctccggc tccgagacgg tgctgggtac ggaagtgaaa 15600

tatgacacgc cgatcacccg caccattacg tctgcaaaca tcgaccgtct gcgctttacc 15660

ttcggtgtac aggcactggt ggaaaccacc tcaaagggtg acaggaatcc gtcggaagtc 15720

cgcctgctgg ttcagataca acgtaacggt ggctgggtga cggaaaaaga catcaccatt 15780

aagggcaaaa ccacctcgca gtatctggcc tcggtggtga tgggtaacct gccgccgcgc 15840

ccgtttaata tccggatgcg caggatgacg ccggacagca ccacagacca gctgcagaac 15900

aaaacgctct ggtcgtcata cactgaaatc atcgatgtga aacagtgcta cccgaacacg 15960

gcactggtcg gcgtgcaggt ggactcggag cagttcggca gccagcaggt gagccgtaat 16020

tatcatctgc gcgggcgtat tctgcaggtg ccgtcgaact ataacccgca gacgcggcaa 16080

tacagcggta tctgggacgg aacgtttaaa ccggcataca gcaacaacat ggcctggtgt 16140

ctgtgggata tgctgaccca tccgcgctac ggcatgggga aacgtcttgg tgcggcggat 16200

gtggataaat gggcgctgta tgtcatcggc cagtactgcg accagtcagt gccggacggc 16260

tttggcggca cggagccgcg catcacctgt aatgcgtacc tgaccacaca gcgtaaggcg 16320

tgggatgtgc tcagcgattt ctgctcggcg atgcgctgta tgccggtatg gaacgggcag 16380

acgctgacgt tcgtgcagga ccgaccgtcg gataagacgt ggacctataa ccgcagtaat 16440

gtggtgatgc cggatgatgg cgcgccgttc cgctacagct tcagcgccct gaaggaccgc 16500

cataatgccg ttgaggtgaa ctggattgac ccgaacaacg gctgggagac ggcgacagag 16560

cttgttgaag atacgcaggc cattgcccgt tacggtcgta atgttacgaa gatggatgcc 16620

tttggctgta ccagccgggg gcaggcacac cgcgccgggc tgtggctgat taaaacagaa 16680

ctgctggaaa cgcagaccgt ggatttcagc gtcggcgcag aagggcttcg ccatgtaccg 16740

ggcgatgtta ttgaaatctg cgatgatgac tatgccggta tcagcaccgg tggtcgtgtg 16800

ctggcggtga acagccagac ccggacgctg acgctcgacc gtgaaatcac gctgccatcc 16860

tccggtaccg cgctgataag cctggttgac ggaagtggca atccggtcag cgtggaggtt 16920

cagtccgtca ccgacggcgt gaaggtaaaa gtgagccgtg ttcctgacgg tgttgctgaa 16980

tacagcgtat gggagctgaa gctgccgacg ctgcgccagc gactgttccg ctgcgtgagt 17040

atccgtgaga acgacgacgg cacgtatgcc atcaccgccg tgcagcatgt gccggaaaaa 17100

gaggccatcg tggataacgg ggcgcacttt gacggcgaac agagtggcac ggtgaatggt 17160

gtcacgccgc cagcggtgca gcacctgacc gcagaagtca ctgcagacag cggggaatat 17220

caggtgctgg cgcgatggga cacaccgaag gtggtgaagg gcgtgagttt cctgctccgt 17280

ctgaccgtaa cagcggacga cggcagtgag cggctggtca gcacggcccg gacgacggaa 17340

accacatacc gcttcacgca actggcgctg gggaactaca ggctgacagt ccgggcggta 17400

aatgcgtggg ggcagcaggg cgatccggcg tcggtatcgt tccggattgc cgcaccggca 17460

gcaccgtcga ggattgagct gacgccgggc tattttcaga taaccgccac gccgcatctt 17520

gccgtttatg acccgacggt acagtttgag ttctggttct cggaaaagca gattgcggat 17580

atcagacagg ttgaaaccag cacgcgttat cttggtacgg cgctgtactg gatagccgcc 17640

agtatcaata tcaaaccggg ccatgattat tacttttata tccgcagtgt gaacaccgtt 17700

ggcaaatcgg cattcgtgga ggccgtcggt cgggcgagcg atgatgcgga aggttacctg 17760

gattttttca aaggcaagat aaccgaatcc catctcggca aggagctgct ggaaaaagtc 17820

gagctgacgg aggataacgc cagcagactg gaggagtttt cgaaagagtg gaaggatgcc 17880

agtgataagt ggaatgccat gtgggctgtc aaaattgagc agaccaaaga cggcaaacat 17940

tatgtcgcgg gtattggcct cagcatggag gacacggagg aaggcaaact gagccagttt 18000

ctggttgccg ccaatcgtat cgcatttatt gacccggcaa acgggaatga aacgccgatg 18060

tttgtggcgc agggcaacca gatattcatg aacgacgtgt tcctgaagcg cctgacggcc 18120

cccaccatta ccagcggcgg caatcctccg gccttttccc tgacaccgga cggaaagctg 18180

accgctaaaa atgcggatat cagtggcagt gtgaatgcga actccgggac gctcagtaat 18240

gtgacgatag ctgaaaactg tacgataaac ggtacgctga gggcggaaaa aatcgtcggg 18300

gacattgtaa aggcggcgag cgcggctttt ccgcgccagc gtgaaagcag tgtggactgg 18360

ccgtcaggta cccgtactgt caccgtgacc gatgaccatc cttttgatcg ccagatagtg 18420

gtgcttccgc tgacgtttcg cggaagtaag cgtactgtca gcggcaggac aacgtattcg 18480

atgtgttatc tgaaagtact gatgaacggt gcggtgattt atgatggcgc ggcgaacgag 18540

gcggtacagg tgttctcccg tattgttgac atgccagcgg gtcggggaaa cgtgatcctg 18600

acgttcacgc ttacgtccac acggcattcg gcagatattc cgccgtatac gtttgccagc 18660

gatgtgcagg ttatggtgat taagaaacag gcgctgggca tcagcgtggt ctgagtgtgt 18720

tacagaggtt cgtccgggaa cgggcgtttt attataaaac agtgagaggt gaacgatgcg 18780

taatgtgtgt attgccgttg ctgtctttgc cgcacttgcg gtgacagtca ctccggcccg 18840

tgcggaaggt ggacatggta cgtttacggt gggctatttt caagtgaaac cgggtacatt 18900

gccgtcgttg tcgggcgggg ataccggtgt gagtcatctg aaagggatta acgtgaagta 18960

ccgttatgag ctgacggaca gtgtgggggt gatggcttcc ctggggttcg ccgcgtcgaa 19020

aaagagcagc acagtgatga ccggggagga tacgtttcac tatgagagcc tgcgtggacg 19080

ttatgtgagc gtgatggccg gaccggtttt acaaatcagt aagcaggtca gtgcgtacgc 19140

catggccgga gtggctcaca gtcggtggtc cggcagtaca atggattacc gtaagacgga 19200

aatcactccc gggtatatga aagagacgac cactgccagg gacgaaagtg caatgcggca 19260

tacctcagtg gcgtggagtg caggtataca gattaatccg gcagcgtccg tcgttgttga 19320

tattgcttat gaaggctccg gcagtggcga ctggcgtact gacggattca tcgttggggt 19380

cggttataaa ttctgattag ccaggtaaca cagtgttatg acagcccgcc ggaaccggtg 19440

ggcttttttg tggggtgaat atggcagtaa agatttcagg agtcctgaaa gacggcacag 19500

gaaaaccggt acagaactgc accattcagc tgaaagccag acgtaacagc accacggtgg 19560

tggtgaacac ggtgggctca gagaatccgg atgaagccgg gcgttacagc atggatgtgg 19620

agtacggtca gtacagtgtc atcctgcagg ttgacggttt tccaccatcg cacgccggga 19680

ccatcaccgt gtatgaagat tcacaaccgg ggacgctgaa tgattttctc tgtgccatga 19740

cggaggatga tgcccggccg gaggtgctgc gtcgtcttga actgatggtg gaagaggtgg 19800

cgcgtaacgc gtccgtggtg gcacagagta cggcagacgc gaagaaatca gccggcgatg 19860

ccagtgcatc agctgctcag gtcgcggccc ttgtgactga tgcaactgac tcagcacgcg 19920

ccgccagcac gtccgccgga caggctgcat cgtcagctca ggaagcgtcc tccggcgcag 19980

aagcggcatc agcaaaggcc actgaagcgg aaaaaagtgc cgcagccgca gagtcctcaa 20040

aaaacgcggc ggccaccagt gccggtgcgg cgaaaacgtc agaaacgaat gctgcagcgt 20100

cacaacaatc agccgccacg tctgcctcca ccgcggccac gaaagcgtca gaggccgcca 20160

cttcagcacg agatgcggtg gcctcaaaag aggcagcaaa atcatcagaa acgaacgcat 20220

catcaagtgc cggtcgtgca gcttcctcgg caacggcggc agaaaattct gccagggcgg 20280

caaaaacgtc cgagacgaat gccaggtcat ctgaaacagc agcggaacgg agcgcctctg 20340

ccgcggcaga cgcaaaaaca gcggcggcgg ggagtgcgtc aacggcatcc acgaaggcga 20400

cagaggctgc gggaagtgcg gtatcagcat cgcagagcaa aagtgcggca gaagcggcgg 20460

caatacgtgc aaaaaattcg gcaaaacgtg cagaagatat agcttcagct gtcgcgcttg 20520

aggatgcgga cacaacgaga aaggggatag tgcagctcag cagtgcaacc aacagcacgt 20580

ctgaaacgct tgctgcaacg ccaaaggcgg ttaaggtggt aatggatgaa acgaacagaa 20640

aagcccactg gacagtccgg cactgaccgg aacgccaaca gcaccaaccg cgctcagggg 20700

aacaaacaat acccagattg cgaacaccgc ttttgtactg gccgcgattg cagatgttat 20760

cgacgcgtca cctgacgcac tgaatacgct gaatgaactg gccgcagcgc tcgggaatga 20820

tccagatttt gctaccacca tgactaacgc gcttgcgggt aaacaaccga agaatgcgac 20880

actgacggcg ctggcagggc tttccacggc gaaaaataaa ttaccgtatt ttgcggaaaa 20940

tgatgccgcc agcctgactg aactgactca ggttggcagg gatattctgg caaaaaattc 21000

cgttgcagat gttcttgaat accttggggc cggtgagaat tcggcctttc cggcaggtgc 21060

gccgatcccg tggccatcag atatcgttcc gtctggctac gtcctgatgc aggggcaggc 21120

gtttgacaaa tcagcctacc caaaacttgc tgtcgcgtat ccatcgggtg tgcttcctga 21180

tatgcgaggc tggacaatca aggggaaacc cgccagcggt cgtgctgtat tgtctcagga 21240

acaggatgga attaagtcgc acacccacag tgccagtgca tccggtacgg atttggggac 21300

gaaaaccaca tcgtcgtttg attacgggac gaaaacaaca ggcagtttcg attacggcac 21360

caaatcgacg aataacacgg gggctcatgc tcacagtctg agcggttcaa caggggccgc 21420

gggtgctcat gcccacacaa gtggtttaag gatgaacagt tctggctgga gtcagtatgg 21480

aacagcaacc attacaggaa gtttatccac agttaaagga accagcacac agggtattgc 21540

ttatttatcg aaaacggaca gtcagggcag ccacagtcac tcattgtccg gtacagccgt 21600

gagtgccggt gcacatgcgc atacagttgg tattggtgcg caccagcatc cggttgttat 21660

cggtgctcat gcccattctt tcagtattgg ttcacacgga cacaccatca ccgttaacgc 21720

tgcgggtaac gcggaaaaca ccgtcaaaaa cattgcattt aactatattg tgaggcttgc 21780

ataatggcat tcagaatgag tgaacaacca cggaccataa aaatttataa tctgctggcc 21840

ggaactaatg aatttattgg tgaaggtgac gcatatattc cgcctcatac cggtctgcct 21900

gcaaacagta ccgatattgc accgccagat attccggctg gctttgtggc tgttttcaac 21960

agtgatgagg catcgtggca tctcgttgaa gaccatcggg gtaaaaccgt ctatgacgtg 22020

gcttccggcg acgcgttatt tatttctgaa ctcggtccgt taccggaaaa ttttacctgg 22080

ttatcgccgg gaggggaata tcagaagtgg aacggcacag cctgggtgaa ggatacggaa 22140

gcagaaaaac tgttccggat ccgggaggcg gaagaaacaa aaaaaagcct gatgcaggta 22200

gccagtgagc atattgcgcc gcttcaggat gctgcagatc tggaaattgc aacgaaggaa 22260

gaaacctcgt tgctggaagc ctggaagaag tatcgggtgt tgctgaaccg tgttgataca 22320

tcaactgcac ctgatattga gtggcctgct gtccctgtta tggagtaa 22368

<210> 8

<211> 20330

<212> DNA

<213> bacteriophage lambda

<400> 8

tcaaggtccc taaattaata cgactcacta tagggagata ggggccttta cgattattac 60

tttaagattt aactctaaga ggaatcttta ttatgttaac acctattaac caattactta 120

agaaccctaa cgatattcca gatgtacctc gtgcaaccgc tgagtatcta caggttcgat 180

tcaactatgc gtacctcgaa gcgtctggtc atataggact tatgcgtgct aatggttgta 240

gtgaggccca catcttgggt ttcattcagg gcctacagta tgcctctaac gtcattgacg 300

agattgagtt acgcaaggaa caactaagag atgatgggga ggattgacac tatgtgtttc 360

tcaccgaaaa ttaaaactcc gaagatggat accaatcaga ttcgagccgt tgagccagcg 420

cctctgaccc aagaagtgtc aagcgtggag ttcggtgggt cttctgatga gacggatacc 480

gagggcaccg aagtgtctgg acgcaaaggc ctcaaggtcg aacgtgatga ttccgtagcg 540

aagtctaaag ccagcggcaa tggctccgct cgtatgaaat cttccatccg taagtccgca 600

tttggaggta agaagtgatg tctgagttca catgtgtgga ggctaagagt cgcttccgtg 660

caatccggtg gactgtggaa caccttgggt tgcctaaagg attcgaagga cactttgtgg 720

gctacagcct ctacgtagac gaagtgatgg acatgtctgg ttgccgtgaa gagtacattc 780

tggactctac cggaaaacat gtagcgtact tcgcgtggtg cgtaagctgt gacattcacc 840

acaaaggaga cattctggat gtaacgtccg ttgtcattaa tcctgaggca gactctaagg 900

gcttacagcg attcctagcg aaacgcttta agtaccttgc ggaactccac gattgcgatt 960

gggtgtctcg ttgtaagcat gaaggcgaga caatgcgtgt atactttaag gaggtataag 1020

ttatgggtaa gaaagttaag aaggccgtga agaaagtcac caagtccgtt aagaaagtcg 1080

ttaaggaagg ggctcgtccg gttaaacagg ttgctggcgg tctagctggt ctggctggtg 1140

gtactggtga agcacagatg gtggaagtac cacaagctgc cgcacagatt gttgacgtac 1200

ctgagaaaga ggtttccact gaggacgaag cacagacaga aagcggacgc aagaaagctc 1260

gtgctggcgg taagaaatcc ttgagtgtag cccgtagctc cggtggcggt atcaacattt 1320

aatcaggagg ttatcgtgga agactgcatt gaatggaccg gaggtgtcaa ctctaagggt 1380

tatggtcgta agtgggttaa tggtaaactt gtgactccac ataggcacat ctatgaggag 1440

acatatggtc cagttccaac aggaattgtg gtgatgcata tctgcgataa ccctaggtgc 1500

tataacataa agcaccttac gcttggaact ccaaaggata attccgagga catggttacc 1560

aaaggtagac aggctaaagg agaggaacta agcaagaaac ttacagagtc agacgttctc 1620

gctatacgct cttcaacctt aagccaccgc tccttaggag aactgtatgg agtcagtcaa 1680

tcaaccataa cgcgaatact acagcgtaag acatggagac acatttaatg gctgagaaac 1740

gaacaggact tgcggaggat ggcgcaaagt ctgtctatga gcgtttaaag aacgaccgtg 1800

ctccctatga gacacgcgct cagaattgcg ctcaatatac catcccatca ttgttcccta 1860

aggactccga taacgcctct acagattatc aaactccgtg gcaagccgtg ggcgctcgtg 1920

gtctgaacaa tctagcctct aagctcatgc tggctctatt ccctatgcag acttggatgc 1980

gacttactat atctgaatat gaagcaaagc agttactgag cgaccccgat ggactcgcta 2040

aggtcgatga gggcctctcg atggtagagc gtatcatcat gaactacatt gagtctaaca 2100

gttaccgcgt gactctcttt gaggctctca aacagttagt cgtagctggt aacgtcctgc 2160

tgtacctacc ggaaccggaa gggtcaaact ataatcccat gaagctgtac cgattgtctt 2220

cttatgtggt ccaacgagac gcattcggca acgttctgca aatggtgact cgtgaccaga 2280

tagcttttgg tgctctccct gaggacatcc gtaaggctgt agaaggtcaa ggtggtgaga 2340

agaaagctga tgagacaatc gacgtgtaca ctcacatcta tctggatgag gactcaggtg 2400

aatacctccg atacgaagag gtcgagggta tggaagtcca aggctccgat gggacttatc 2460

ctaaagaggc ttgcccatac atcccgattc ggatggtcag actagatggt gaatcctacg 2520

gtcgttcgta cattgaggaa tacttaggtg acttacggtc ccttgaaaat ctccaagagg 2580

ctatcgtcaa gatgtccatg attagctcta aggttatcgg cttagtgaat cctgctggta 2640

tcacccagcc acgccgactg accaaagctc agactggtga cttcgttact ggtcgtccag 2700

aagacatctc gttcctccaa ctggagaagc aagcagactt tactgtagct aaagccgtaa 2760

gtgacgctat cgaggctcgc ctttcgtttg cctttatgtt gaactctgcg gttcagcgta 2820

caggtgaacg tgtgaccgcc gaagagattc ggtatgtagc ttctgaactt gaagatactt 2880

taggtggtgt ctactctatc ctttctcaag aattacaatt gcctctggta cgagtgctct 2940

tgaagcaact acaagccacg caacagattc ctgagttacc taaggaagcc gtagagccaa 3000

ccattagtac aggtctggaa gcaattggtc gaggacaaga ccttgataag ctggagcggt 3060

gtgtcactgc gtgggctgca ctggcaccta tgcgggacga ccctgatatt aaccttgcga 3120

tgattaagtt acgtattgcc aacgctatcg gtattgacac ttctggtatt ctactcaccg 3180

aagaacagaa gcaacagaag atggcccaac agtctatgca aatgggtatg gataatggtg 3240

ctgctgcgct ggctcaaggt atggctgcac aagctacagc ttcacctgag gctatggctg 3300

ctgccgctga ttccgtaggt ttacagccgg gaatttaata cgactcacta tagggagacc 3360

tcatctttga aatgagcgat gacaagaggt tggagtcctc ggtcttcctg tagttcaact 3420

ttaaggagac aataataatg gctgaatcta atgcagacgt atatgcatct tttggcgtga 3480

actccgctgt gatgtctggt ggttccgttg aggaacatga gcagaacatg ctggctcttg 3540

atgttgctgc ccgtgatggc gatgatgcaa tcgagttagc gtcagacgaa gtggaaacag 3600

aacgtgacct gtatgacaac tctgacccgt tcggtcaaga ggatgacgaa ggccgcattc 3660

aggttcgtat cggtgatggc tctgagccga ccgatgtgga cactggagaa gaaggcgttg 3720

agggcaccga aggttccgaa gagtttaccc cactgggcga gactccagaa gaactggtag 3780

ctgcctctga gcaacttggt gagcacgaag agggcttcca agagatgatt aacattgctg 3840

ctgagcgtgg catgagtgtc gagaccattg aggctatcca gcgtgagtac gaggagaacg 3900

aagagttgtc cgccgagtcc tacgctaagc tggctgaaat tggctacacg aaggctttca 3960

ttgactcgta tatccgtggt caagaagctc tggtggagca gtacgtaaac agtgtcattg 4020

agtacgctgg tggtcgtgaa cgttttgatg cactgtataa ccaccttgag acgcacaacc 4080

ctgaggctgc acagtcgctg gataatgcgt tgaccaatcg tgacttagcg accgttaagg 4140

ctatcatcaa cttggctggt gagtctcgcg ctaaggcgtt cggtcgtaag ccaactcgta 4200

gtgtgactaa tcgtgctatt ccggctaaac ctcaggctac caagcgtgaa ggctttgcgg 4260

accgtagcga gatgattaaa gctatgagtg accctcggta tcgcacagat gccaactatc 4320

gtcgtcaagt cgaacagaaa gtaatcgatt cgaacttctg atagacttcg aaattaatac 4380

gactcactat agggagacca caacggtttc cctctagaaa taattttgtt taactttaag 4440

aaggagatat acatatggct agcatgactg gtggacagca aatgggtact aaccaaggta 4500

aaggtgtagt tgctgctgga gataaactgg cgttgttctt gaaggtattt ggcggtgaag 4560

tcctgactgc gttcgctcgt acctccgtga ccacttctcg ccacatggta cgttccatct 4620

ccagcggtaa atccgctcag ttccctgttc tgggtcgcac tcaggcagcg tatctggctc 4680

cgggcgagaa cctcgacgat aaacgtaagg acatcaaaca caccgagaag gtaatcacca 4740

ttgacggtct cctgacggct gacgttctga tttatgatat tgaggacgcg atgaaccact 4800

acgacgttcg ctctgagtat acctctcagt tgggtgaatc tctggcgatg gctgcggatg 4860

gtgcggttct ggctgagatt gccggtctgt gtaacgtgga aagcaaatat aatgagaaca 4920

tcgagggctt aggtactgct accgtaattg agaccactca gaacaaggcc gcacttaccg 4980

accaagttgc gctgggtaag gagattattg cggctctgac taaggctcgt gcggctctga 5040

ccaagaacta tgttccggct gctgaccgtg tgttctactg tgacccagat agctactctg 5100

cgattctggc agcactgatg ccgaacgcag caaactacgc tgctctgatt gaccctgaga 5160

agggttctat ccgcaacgtt atgggctttg aggttgtaga agttccgcac ctcaccgctg 5220

gtggtgctgg taccgctcgt gagggcacta ctggtcagaa gcacgtcttc cctgccaata 5280

aaggtgaggg taatgtcaag gttgctaagg acaacgttat cggcctgttc atgcaccgct 5340

ctgcggtagg tactgttaag ctgcgtgact tggctctgga gcgcgctcgc cgtgctaact 5400

tccaagcgga ccagattatc gctaagtacg caatgggcca cggtggtctt cgcccagaag 5460

ctgctggtgc agtggttttc aaagtggagt aatgctgggg gtggcctcaa cggtcgctgc 5520

tagtcccgaa gaggcgagtg ttacttcaac agaagaaacc ttaacgccag cacaggaggc 5580

cgcacgcacc cgcgctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc 5640

tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct 5700

gaaaggagga actatatgcg ctcatacgat atgaacgttg agactgccgc tgagttatca 5760

gctgtgaacg acattctggc gtctatcggt gaacctccgg tatcaacgct ggaaggtgac 5820

gctaacgcag atgcagcgaa cgctcggcgt attctcaaca agattaaccg acagattcaa 5880

tctcgtggat ggacgttcaa cattgaggaa ggcataacgc tactacctga tgtttactcc 5940

aacctgattg tatacagtga cgactattta tccctaatgt ctacttccgg tcaatccatc 6000

tacgttaacc gaggtggcta tgtgtatgac cgaacgagtc aatcagaccg ctttgactct 6060

ggtattactg tgaacattat tcgtctccgc gactacgatg agatgcctga gtgcttccgt 6120

tactggattg tcaccaaggc ttcccgtcag ttcaacaacc gattctttgg ggcaccggaa 6180

gtagagggtg tactccaaga agaggaagat gaggctagac gtctctgcat ggagtatgag 6240

atggactacg gtgggtacaa tatgctggat ggagatgcgt tcacttctgg tctactgact 6300

cgctaacatt aataaataag gaggctctaa tggcactcat tagccaatca atcaagaact 6360

tgaagggtgg tatcagccaa cagcctgaca tccttcgtta tccagaccaa gggtcacgcc 6420

aagttaacgg ttggtcttcg gagaccgagg gcctccaaaa gcgtccacct cttgttttct 6480

taaatacact tggagacaac ggtgcgttag gtcaagctcc gtacatccac ctgattaacc 6540

gagatgagca cgaacagtat tacgctgtgt tcactggtag cggaatccga gtgttcgacc 6600

tttctggtaa cgagaagcaa gttaggtatc ctaacggttc caactacatc aagaccgcta 6660

atccacgtaa cgacctgcga atggttactg tagcagacta tacgttcatc gttaaccgta 6720

acgttgttgc acagaagaac acaaagtctg tcaacttacc gaattacaac cctaatcaag 6780

acggattgat taacgttcgt ggtggtcagt atggtaggga actaattgta cacattaacg 6840

gtaaagacgt tgcgaagtat aagataccag atggtagtca acctgaacac gtaaacaata 6900

cggatgccca atggttagct gaagagttag ccaagcagat gcgcactaac ttgtctgatt 6960

ggactgtaaa tgtagggcaa gggttcatcc atgtgaccgc acctagtggt caacagattg 7020

actccttcac gactaaagat ggctacgcag accagttgat taaccctgtg acccactacg 7080

ctcagtcgtt ctctaagctg ccacctaatg ctcctaacgg ctacatggtg aaaatcgtag 7140

gggacgcctc taagtctgcc gaccagtatt acgttcggta tgacgctgag cggaaagttt 7200

ggactgagac tttaggttgg aacactgagg accaagttct atgggaaacc atgccacacg 7260

ctcttgtgcg agccgctgac ggtaatttcg acttcaagtg gcttgagtgg tctcctaagt 7320

cttgtggtga cgttgacacc aacccttggc cttcttttgt tggttcaagt attaacgatg 7380

tgttcttctt ccgtaaccgc ttaggattcc ttagtgggga gaacatcata ttgagtcgta 7440

cagccaaata cttcaacttc taccctgcgt ccattgcgaa ccttagtgat gacgacccta 7500

tagacgtagc tgtgagtacc aaccgaatag caatccttaa gtacgccgtt ccgttctcag 7560

aagagttact catctggtcc gatgaagcac aattcgtcct gactgcctcg ggtactctca 7620

catctaagtc ggttgagttg aacctaacga cccagtttga cgtacaggac cgagcgagac 7680

cttttgggat tgggcgtaat gtctactttg ctagtccgag gtccagcttc acgtccatcc 7740

acaggtacta cgctgtgcag gatgtcagtt ccgttaagaa tgctgaggac attacatcac 7800

acgttcctaa ctacatccct aatggtgtgt tcagtatttg cggaagtggt acggaaaact 7860

tctgttcggt actatctcac ggggacccta gtaaaatctt catgtacaaa ttcctgtacc 7920

tgaacgaaga gttaaggcaa cagtcgtggt ctcattggga ctttggggaa aacgtacagg 7980

ttctagcttg tcagagtatc agctcagata tgtatgtgat tcttcgcaat gagttcaata 8040

cgttcctagc tagaatctct ttcactaaga acgccattga cttacaggga gaaccctatc 8100

gtgcctttat ggacatgaag attcgataca cgattcctag tggaacatac aacgatgaca 8160

cattcactac ctctattcat attccaacaa tttatggtgc aaacttcggg aggggcaaaa 8220

tcactgtatt ggagcctgat ggtaagataa ccgtgtttga gcaacctacg gctgggtgga 8280

atagcgaccc ttggctgaga ctcagcggta acttggaggg acgcatggtg tacattgggt 8340

tcaacattaa cttcgtatat gagttctcta agttcctcat caagcagact gccgacgacg 8400

ggtctacctc cacggaagac attgggcgct tacagttacg ccgagcgtgg gttaactacg 8460

agaactctgg tacgtttgac atttatgttg agaaccaatc gtctaactgg aagtacacaa 8520

tggctggtgc ccgattaggc tctaacactc tgagggctgg gagactgaac ttagggaccg 8580

gacaatatcg attccctgtg gttggtaacg ccaagttcaa cactgtatac atcttgtcag 8640

atgagactac ccctctgaac atcattgggt gtggctggga aggtaactac ttacggagaa 8700

gttccggtat ttaattaaat attctccctg tggtggctcg aaattaatac gactcactat 8760

agggagaaca atacgactac gggagggttt tcttatgatg actataagac ctactaaaag 8820

tacagacttt gaggtattca ctccggctca ccatgacatt cttgaagcta aggctgctgg 8880

tattgagccg agtttccctg atgcttccga gtgtgtcacg ttgagcctct atgggttccc 8940

tctagctatc ggtggtaact gcggggacca gtgctggttc gttacgagcg accaagtgtg 9000

gcgacttagt ggaaaggcta agcgaaagtt ccgtaagtta atcatggagt atcgcgataa 9060

gatgcttgag aagtatgata ctctttggaa ttacgtatgg gtaggcaata cgtcccacat 9120

tcgtttcctc aagactatcg gtgcggtatt ccatgaagag tacacacgag atggtcaatt 9180

tcagttattt acaatcacga aaggaggata accatatgtg ttgggcagcc gcaataccta 9240

tcgctatatc tggcgctcag gctatcagtg gtcagaacgc tcaggccaaa atgattgccg 9300

ctcagaccgc tgctggtcgt cgtcaagcta tggaaatcat gaggcagacg aacatccaga 9360

atgctgacct atcgttgcaa gctcgaagta aacttgagga agcgtccgcc gagttgacct 9420

cacagaacat gcagaaggtc caagctattg ggtctatccg agcggctatc ggagagagta 9480

tgcttgaagg ttcctcaatg gaccgcatta agcgagtcac agaaggacag ttcattcggg 9540

aagccaatat ggtaactgag aactatcgcc gtgactacca agcaatcttc gcacagcaac 9600

ttggtggtac tcaaagtgct gcaagtcaga ttgacgaaat ctataagagc gaacagaaac 9660

agaagagtaa gctacagatg gttctggacc cactggctat catggggtct tccgctgcga 9720

gtgcttacgc atccggtgcg ttcgactcta agtccacaac taaggcacct attgttgccg 9780

ctaaaggaac caagacgggg aggtaatgag ctatgagtaa aattgaatct gcccttcaag 9840

cggcacaacc gggactctct cggttacgtg gtggtgctgg aggtatgggc tatcgtgcag 9900

caaccactca ggccgaacag ccaaggtcaa gcctattgga caccattggt cggttcgcta 9960

aggctggtgc cgatatgtat accgctaagg aacaacgagc acgagaccta gctgatgaac 10020

gctctaacga gattatccgt aagctgaccc ctgagcaacg tcgagaagct ctcaacaacg 10080

ggacccttct gtatcaggat gacccatacg ctatggaagc actccgagtc aagactggtc 10140

gtaacgctgc gtatcttgtg gacgatgacg ttatgcagaa gataaaagag ggtgtcttcc 10200

gtactcgcga agagatggaa gagtatcgcc atagtcgcct tcaagagggc gctaaggtat 10260

acgctgagca gttcggcatc gaccctgagg acgttgatta tcagcgtggt ttcaacgggg 10320

acattaccga gcgtaacatc tcgctgtatg gtgcgcatga taacttcttg agccagcaag 10380

ctcagaaggg cgctatcatg aacagccgag tggaactcaa cggtgtcctt caagaccctg 10440

atatgctgcg tcgtccagac tctgctgact tctttgagaa gtatatcgac aacggtctgg 10500

ttactggcgc aatcccatct gatgctcaag ccacacagct tataagccaa gcgttcagtg 10560

acgcttctag ccgtgctggt ggtgctgact tcctgatgcg agtcggtgac aagaaggtaa 10620

cacttaacgg agccactacg acttaccgag agttgattgg tgaggaacag tggaacgctc 10680

tcatggtcac agcacaacgt tctcagtttg agactgacgc gaagctgaac gagcagtatc 10740

gcttgaagat taactctgcg ctgaaccaag aggacccaag gacagcttgg gagatgcttc 10800

aaggtatcaa ggctgaacta gataaggtcc aacctgatga gcagatgaca ccacaacgtg 10860

agtggctaat ctccgcacag gaacaagttc agaatcagat gaacgcatgg acgaaagctc 10920

aggccaaggc tctggacgat tccatgaagt caatgaacaa acttgacgta atcgacaagc 10980

aattccagaa gcgaatcaac ggtgagtggg tctcaacgga ttttaaggat atgccagtca 11040

acgagaacac tggtgagttc aagcatagcg atatggttaa ctacgccaat aagaagctcg 11100

ctgagattga cagtatggac attccagacg gtgccaagga tgctatgaag ttgaagtacc 11160

ttcaagcgga ctctaaggac ggagcattcc gtacagccat cggaaccatg gtcactgacg 11220

ctggtcaaga gtggtctgcc gctgtgatta acggtaagtt accagaacga accccagcta 11280

tggatgctct gcgcagaatc cgcaatgctg accctcagtt gattgctgcg ctatacccag 11340

accaagctga gctattcctg acgatggaca tgatggacaa gcagggtatt gaccctcagg 11400

ttattcttga tgccgaccga ctgactgtta agcggtccaa agagcaacgc tttgaggatg 11460

ataaagcatt cgagtctgca ctgaatgcat ctaaggctcc tgagattgcc cgtatgccag 11520

cgtcactgcg cgaatctgca cgtaagattt atgactccgt taagtatcgc tcggggaacg 11580

aaagcatggc tatggagcag atgaccaagt tccttaagga atctacctac acgttcactg 11640

gtgatgatgt tgacggtgat accgttggtg tgattcctaa gaatatgatg caggttaact 11700

ctgacccgaa atcatgggag caaggtcggg atattctgga ggaagcacgt aagggaatca 11760

ttgcgagcaa cccttggata accaataagc aactgaccat gtattctcaa ggtgactcca 11820

tttaccttat ggacaccaca ggtcaagtca gagtccgata cgacaaagag ttactctcga 11880

aggtctggag tgagaaccag aagaaactcg aagagaaagc tcgtgagaag gctctggctg 11940

atgtgaacaa gcgagcacct atagttgccg ctacgaaggc ccgtgaagct gctgctaaac 12000

gagtccgaga gaaacgtaaa cagactccta agttcatcta cggacgtaag gagtaactaa 12060

aggctacata aggaggccct aaatggataa gtacgataag aacgtaccaa gtgattatga 12120

tggtctgttc caaaaggctg ctgatgccaa cggggtctct tatgaccttt tacgtaaagt 12180

cgcttggaca gaatcacgat ttgtgcctac agcaaaatct aagactggac cattaggcat 12240

gatgcaattt accaaggcaa ccgctaaggc cctcggtctg cgagttaccg atggtccaga 12300

cgacgaccga ctgaaccctg agttagctat taatgctgcc gctaagcaac ttgcaggtct 12360

ggtagggaag tttgatggcg atgaactcaa agctgccctt gcgtacaacc aaggcgaggg 12420

acgcttgggt aatccacaac ttgaggcgta ctctaaggga gacttcgcat caatctctga 12480

ggagggacgt aactacatgc gtaaccttct ggatgttgct aagtcaccta tggctggaca 12540

gttggaaact tttggtggca taaccccaaa gggtaaaggc attccggctg aggtaggatt 12600

ggctggaatt ggtcacaagc agaaagtaac acaggaactt cctgagtcca caagttttga 12660

cgttaagggt atcgaacagg aggctacggc gaaaccattc gccaaggact tttgggagac 12720

ccacggagaa acacttgacg agtacaacag tcgttcaacc ttcttcggat tcaaaaatgc 12780

tgccgaagct gaactctcca actcagtcgc tgggatggct ttccgtgctg gtcgtctcga 12840

taatggtttt gatgtgttta aagacaccat tacgccgact cgctggaact ctcacatctg 12900

gactccagag gagttagaga agattcgaac agaggttaag aaccctgcgt acatcaacgt 12960

tgtaactggt ggttcccctg agaacctcga tgacctcatt aaattggcta acgagaactt 13020

tgagaatgac tcccgcgctg ccgaggctgg cctaggtgcc aaactgagtg ctggtattat 13080

tggtgctggt gtggacccgc ttagctatgt tcctatggtc ggtgtcactg gtaagggctt 13140

taagttaatc aataaggctc ttgtagttgg tgccgaaagt gctgctctga acgttgcatc 13200

cgaaggtctc cgtacctccg tagctggtgg tgacgcagac tatgcgggtg ctgccttagg 13260

tggctttgtg tttggcgcag gcatgtctgc aatcagtgac gctgtagctg ctggactgaa 13320

acgcagtaaa ccagaagctg agttcgacaa tgagttcatc ggtcctatga tgcgattgga 13380

agcccgtgag acagcacgaa acgccaactc tgcggacctc tctcggatga acactgagaa 13440

catgaagttt gaaggtgaac ataatggtgt cccttatgag gacttaccaa cagagagagg 13500

tgccgtggtg ttacatgatg gctccgttct aagtgcaagc aacccaatca accctaagac 13560

tctaaaagag ttctccgagg ttgaccctga gaaggctgcg cgaggaatca aactggctgg 13620

gttcaccgag attggcttga agaccttggg gtctgacgat gctgacatcc gtagagtggc 13680

tatcgacctc gttcgctctc ctactggtat gcagtctggt gcctcaggta agttcggtgc 13740

aacagcttct gacatccatg agagacttca tggtactgac cagcgtactt ataatgactt 13800

gtacaaagca atgtctgacg ctatgaaaga ccctgagttc tctactggcg gcgctaagat 13860

gtcccgtgaa gaaactcgat acactatcta ccgtagagcg gcactagcta ttgagcgtcc 13920

agaactacag aaggcactca ctccgtctga gagaatcgtt atggacatca ttaagcgtca 13980

ctttgacacc aagcgtgaac ttatggaaaa cccagcaata ttcggtaaca caaaggctgt 14040

gagtatcttc cctgagagtc gccacaaagg tacttacgtt cctcacgtat atgaccgtca 14100

tgccaaggcg ctgatgattc aacgctacgg tgccgaaggt ttgcaggaag ggattgcccg 14160

ctcatggatg aacagctacg tctccagacc tgaggtcaag gccagagtcg atgagatgct 14220

taaggaatta cacggggtga aggaagtaac accagagatg gtagagaagt acgctatgga 14280

taaggcttat ggtatctccc actcagacca gttcaccaac agttccataa tagaagagaa 14340

cattgagggc ttagtaggta tcgagaataa ctcattcctt gaggcacgta acttgtttga 14400

ttcggaccta tccatcacta tgccagacgg acagcaattc tcagtgaatg acctaaggga 14460

cttcgatatg ttccgcatca tgccagcgta tgaccgccgt gtcaatggtg acatcgccat 14520

catggggtct actggtaaaa ccactaagga acttaaggat gagattttgg ctctcaaagc 14580

gaaagctgag ggagacggta agaagactgg cgaggtacat gctttaatgg ataccgttaa 14640

gattcttact ggtcgtgcta gacgcaatca ggacactgtg tgggaaacct cactgcgtgc 14700

catcaatgac ctagggttct tcgctaagaa cgcctacatg ggtgctcaga acattacgga 14760

gattgctggg atgattgtca ctggtaacgt tcgtgctcta gggcatggta tcccaattct 14820

gcgtgataca ctctacaagt ctaaaccagt ttcagctaag gaactcaagg aactccatgc 14880

gtctctgttc gggaaggagg tggaccagtt gattcggcct aaacgtgctg acattgtgca 14940

gcgcctaagg gaagcaactg ataccggacc tgccgtggcg aacatcgtag ggaccttgaa 15000

gtattcaaca caggaactgg ctgctcgctc tccgtggact aagctactga acggaaccac 15060

taactacctt ctggatgctg cgcgtcaagg tatgcttggg gatgttatta gtgccaccct 15120

aacaggtaag actacccgct gggagaaaga aggcttcctt cgtggtgcct ccgtaactcc 15180

tgagcagatg gctggcatca agtctctcat caaggaacat atggtacgcg gtgaggacgg 15240

gaagtttacc gttaaggaca agcaagcgtt ctctatggac ccacgggcta tggacttatg 15300

gagactggct gacaaggtag ctgatgaggc aatgctgcgt ccacataagg tgtccttaca 15360

ggattcccat gcgttcggag cactaggtaa gatggttatg cagtttaagt ctttcactat 15420

caagtccctt aactctaagt tcctgcgaac cttctatgat ggatacaaga acaaccgagc 15480

gattgacgct gcgctgagca tcatcacctc tatgggtctc gctggtggtt tctatgctat 15540

ggctgcacac gtcaaagcat acgctctgcc taaggagaaa cgtaaggagt acttggagcg 15600

tgcactggac ccaaccatga ttgcccacgc tgcgttatct cgtagttctc aattgggtgc 15660

tcctttggct atggttgacc tagttggtgg tgttttaggg ttcgagtcct ccaagatggc 15720

tcgctctacg attctaccta aggacaccgt gaaggaacgt gacccaaaca aaccgtacac 15780

ctctagagag gtaatgggcg ctatgggttc aaaccttctg gaacagatgc cttcggctgg 15840

ctttgtggct aacgtagggg ctaccttaat gaatgctgct ggcgtggtca actcacctaa 15900

taaagcaacc gagcaggact tcatgactgg tcttatgaac tccacaaaag agttagtacc 15960

gaacgaccca ttgactcaac agcttgtgtt gaagatttat gaggcgaacg gtgttaactt 16020

gagggagcgt aggaaataat acgactcact atagggagag gcgaaataat cttctccctg 16080

tagtctctta gatttacttt aaggaggtca aatggctaac gtaattaaaa ccgttttgac 16140

ttaccagtta gatggctcca atcgtgattt taatatcccg tttgagtatc tagcccgtaa 16200

gttcgtagtg gtaactctta ttggtgtaga ccgaaaggtc cttacgatta atacagacta 16260

tcgctttgct acacgtacta ctatctctct gacaaaggct tggggtccag ccgatggcta 16320

cacgaccatc gagttacgtc gagtaacctc cactaccgac cgattggttg actttacgga 16380

tggttcaatc ctccgcgcgt atgaccttaa cgtcgctcag attcaaacga tgcacgtagc 16440

ggaagaggcc cgtgacctca ctacggatac tatcggtgtc aataacgatg gtcacttgga 16500

tgctcgtggt cgtcgaattg tgaacctagc gaacgccgtg gatgaccgcg atgctgttcc 16560

gtttggtcaa ctaaagacca tgaaccagaa ctcatggcaa gcacgtaatg aagccttaca 16620

gttccgtaat gaggctgaga ctttcagaaa ccaagcggag ggctttaaga acgagtccag 16680

taccaacgct acgaacacaa agcagtggcg cgatgagacc aagggtttcc gagacgaagc 16740

caagcggttc aagaatacgg ctggtcaata cgctacatct gctgggaact ctgcttccgc 16800

tgcgcatcaa tctgaggtaa acgctgagaa ctctgccaca gcatccgcta actctgctca 16860

tttggcagaa cagcaagcag accgtgcgga acgtgaggca gacaagctgg aaaattacaa 16920

tggattggct ggtgcaattg ataaggtaga tggaaccaat gtgtactgga aaggaaatat 16980

tcacgctaac gggcgccttt acatgaccac aaacggtttt gactgtggcc agtatcaaca 17040

gttctttggt ggtgtcacta atcgttactc tgtcatggag tggggagatg agaacggatg 17100

gctgatgtat gttcaacgta gagagtggac aacagcgata ggcggtaaca tccagttagt 17160

agtaaacgga cagatcatca cccaaggtgg agccatgacc ggtcagctaa aattgcagaa 17220

tgggcatgtt cttcaattag agtccgcatc cgacaaggcg cactatattc tatctaaaga 17280

tggtaacagg aataactggt acattggtag agggtcagat aacaacaatg actgtacctt 17340

ccactcctat gtacatggta cgaccttaac actcaagcag gactatgcag tagttaacaa 17400

acacttccac gtaggtcagg ccgttgtggc cactgatggt aatattcaag gtactaagtg 17460

gggaggtaaa tggctggatg cttacctacg tgacagcttc gttgcgaagt ccaaggcgtg 17520

gactcaggtg tggtctggta gtgctggcgg tggggtaagt gtgactgttt cacaggatct 17580

ccgcttccgc aatatctgga ttaagtgtgc caacaactct tggaacttct tccgtactgg 17640

ccccgatgga atctacttca tagcctctga tggtggatgg ttacgattcc aaatacactc 17700

caacggtctc ggattcaaga atattgcaga cagtcgttca gtacctaatg caatcatggt 17760

ggagaacgag taattggtaa atcacaagga aagacgtgta gtccacggat ggactctcaa 17820

ggaggtacaa gtatgtatgg aaaaggataa gagccttatt acattcttag agatgttgga 17880

cactgcgatg gctcagcgta tgcttgcgga cctttcggac catgagcgtc gctctccgca 17940

actctataat gctattaaca aactgttaga ccgccacaag ttccagattg gtaagttgca 18000

gccggatgtt cacatcttag gtggccttgc tggtgctctt gaagagtaca aagagaaagt 18060

cggtgataac ggtcttacgg atgatgatat ttacacatta cagtgatata ctcaaggcag 18120

atagtggtct ttatggatgt cattgtctat acgagatgct cctacgtgaa atctgaaagt 18180

taacgggagg cattgaaatc aagtaaggag gcaatgtgtc tactcaatcc aatcgtaatg 18240

cgctcgtagt ggcgcaactg aaaggagact tcgtggcgtt cctattcgtc ttatggaagg 18300

cgctaaacct accggtgccc actaagtgtc agattgacat ggctaaggtg ctggcgaatg 18360

gagacaacaa gaagttcatc ttacaggctt tccgtggtat cggtaagtcg ttcatcacat 18420

gtgcgttcgt tgtgtggtcc ttatggagag accctcagtt gaagatactt atcgtatcag 18480

cctctaagga gcgtgcagac gctaactcca tctttattaa gaacatcatt gacctgctgc 18540

cattcctatc tgagttaaag ccaagacccg gacagcgtga ctcggtaatc agctttgatg 18600

taggcccagc caatcctgac cactctccta gtgtgaaatc agtaggtatc actggtcagt 18660

taactggtag ccgtgctgac attatcattg cggatgacgt tgagattccg tctaacagcg 18720

caactatggg tgcccgtgag aagctatgga ctctggttca ggagttcgct gcgttactta 18780

aaccgctgcc ttcctctcgc gttatctacc ttggtacacc tcagacagag atgactctct 18840

ataaggaact tgaggataac cgtgggtaca caaccattat ctggcctgct ctgtacccaa 18900

ggacacgtga agagaacctc tattactcac agcgtcttgc tcctatgtta cgcgctgagt 18960

acgatgagaa ccctgaggca cttgctggga ctccaacaga cccagtgcgc tttgaccgtg 19020

atgacctgcg cgagcgtgag ttggaatacg gtaaggctgg ctttacgcta cagttcatgc 19080

ttaaccctaa ccttagtgat gccgagaagt acccgctgag gcttcgtgac gctatcgtag 19140

cggccttaga cttagagaag gccccaatgc attaccagtg gcttccgaac cgtcagaaca 19200

tcattgagga ccttcctaac gttggcctta agggtgatga cctgcatacg taccacgatt 19260

gttccaacaa ctcaggtcag taccaacaga agattctggt cattgaccct agtggtcgcg 19320

gtaaggacga aacaggttac gctgtgctgt acacactgaa cggttacatc taccttatgg 19380

aagctggagg tttccgtgat ggctactccg ataagaccct tgagttactc gctaagaagg 19440

caaagcaatg gggagtccag acggttgtct acgagagtaa cttcggtgac ggtatgttcg 19500

gtaaggtatt cagtcctatc cttcttaaac accacaactg tgcgatggaa gagattcgtg 19560

cccgtggtat gaaagagatg cgtatttgcg atacccttga gccagtcatg cagactcacc 19620

gccttgtaat tcgtgatgag gtcattaggg ccgactacca gtccgctcgt gacgtagacg 19680

gtaagcatga cgttaagtac tcgttgttct accagatgac ccgtatcact cgtgagaaag 19740

gcgctctggc tcatgatgac cgattggatg cccttgcgtt aggcattgag tatctccgtg 19800

agtccatgca gttggattcc gttaaagtag aaggcgaggt tttagcagat tttttagaag 19860

agcatatgat gcgcccaacc gtagcagcaa cccacattat cgaaatgagc gttggtggtg 19920

tggacgttta tagtgaagat gacgaaggct atggcaccag ctttatcgaa tggtaaggac 19980

caacataaag ggaggagact catgttccgc ttattgttga acctactgcg gcatagagtc 20040

acctaccgat ttcttgtggt actttgtgct gcccttgggt acgcatctct tactggagac 20100

ctcagttcac tggagtctgt cgtttgctct atactcactt gtagcgatta gggtcttcct 20160

gaccgactga tggctcaccg agggattcag cggtatgatt gcatcacacc acttcatccc 20220

tatagagtca agtcctaagg tatacccata aagagcctct aatggtctat cctaaggtct 20280

atacctaaag ataggccatc ctatcagtgt cacctaaaga gggtcttaga 20330

<210> 9

<211> 3555

<212> DNA

<213> artificial sequence

<220>

<223> payload pJ23115-GFP T7 cos 2.0

<400> 9

cctttaggga aatatgctaa gttttcaccg taacacgcca catcttgact atatatgtgt 60

agaaactgcc ggaaatcgtc gtggtattct gaccagagcg atgaaaacgt ttcagtttgc 120

tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc 180

attgccatac gaaactccgg atgtgcattc atcaggcggg caagaatgtg aataaaggcc 240

ggataaaact tgtgcttatt tttctttacg gtttttaaaa aggccgtaat atccagctga 300

acggtttggt tataggtgca ctgagcaact gactggaatg cctcaaaatg ttctttacga 360

tgccattgac ttatatcaac tgtagtatat ccagtgattt ttttctccat tttagcttcc 420

ttagcttgcg aaatctcgat aactcaaaaa atagtagtga tcttatttca ttatggtgaa 480

agttgtctta cgtgcaacat tttcgcaaaa agttggcgct tgatttcagt gcaatttatc 540

tcttcaaatg tagcacttta tagctagctc agcccttggt acaatgctag cgttttcatt 600

aaagaggaga aaggaagcca tgagtaaagg tgaggaatta tttactggtg ttgttccgat 660

cttagttgaa ctggacggcg atgttaacgg tcataaattc agtgttcgtg gtgaaggtga 720

aggtgatgca accaacggta agctgaccct gaaattcatc tgcactactg gaaaattacc 780

agtaccgtgg cctactctgg tgactaccct gacctatggt gttcagtgtt tttctcgtta 840

ccctgaccac atgaagcaac atgatttctt caaatctgca atgccggaag gttatgtaca 900

ggagcgcacc atttctttca aagacgatgg cacgtataaa acccgtgcag aggttaaatt 960

tgaaggtgac actctggtga atcgtattga actgaaaggc attgatttca aagaggacgg 1020

caatatttta ggccacaaac tggaatataa cttcaactcc cataacgttt acatcaccgc 1080

agacaaacaa aagaacggta tcaaagctaa cttcaaaatt cgccataacg ttgaagacgg 1140

tagcgtacag ctggcggatc attaccaaca gaacactccg attggagatg ctcctgtttt 1200

actgccggat aaccactacc tgtccaccca gtctaaactg tcgaaggatc cgaacgaaaa 1260

gcgcgaccac atggtgttat tagagttcgt taccgctagt ggtatcacgc acggtatgga 1320

tgaactctac aaataagtca gtttcacctg ttttacgtta aaacccgctt cggcgggttt 1380

ttacttttgg gtttagccga acgccatagt acatgtaggt cgagggtgaa gtacttgctg 1440

acttccttga ggaacacatg atgcgtccta cggttgctgc tacgcatatc attgagatgt 1500

ctgtgggagg agttgatgtg tactctgagg acgatgaggg ttacggtacg tctttcattg 1560

agtggtgatt tatgcattag gactgcatag ggatgcacta tagaccacgg atggtcagtt 1620

ctttaagtta ctgaaaagac acgataaatt aatacgactc actataggga gaggagggac 1680

gaaaggttac tatatagata ctgaatgaat acttatagag tgcataaagt atgcataatg 1740

gtgtacctag agtgacctct aagaatggtg attatattgt attagtatca ccttaactta 1800

aggcgggatc gtcaccctca gcagcgaaag acagctgtcg gtcagagcgt cattgcgaag 1860

ctgagtgtga tcgatgccat cagcgaaggg cccaaactcc gagcgattaa gcgtttgctg 1920

gctgtcacgc ctgcctgttg cttgcttgga cttgcgatgt acgtgctcag ctgtctttcg 1980

ctgctgaggg tgacgatccc gcgagggcct atggagttcc tatagggtcc tttaaaatat 2040

accataaaaa tctgagtgac tatctcacag tgtacggacc taaagttccc ccataggggg 2100

tacctaaagc ccagccaatc acctaaagtc aaccttcggt tgaccttgag ggttccctaa 2160

gggttgggga tgacccttgg gtttgtcttt gggtgttacc ttgagtgtct ctctgtgtcc 2220

ctatctgtta cagtctccta aagtatcctc ctaaagtcac ctcctaacgt agaaatattt 2280

tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc 2340

tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc 2400

tttgaaccga ggtaactggc ttggaggagc gcagtcgcca aaacttgtcc tttcagttta 2460

gccttatccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg 2520

ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg 2580

cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct 2640

acccggaact gagtgtcagg cgtggaatga gacaaactcg gccgtaacag aggaatgaca 2700

ccggcaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc 2760

tggtatcttt atagtcctgt caggtttcgc caccactgat ttgagcgtca gatttcgtga 2820

tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcgaccct ctcacttccc 2880

tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg 2940

ctcgccacag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc 3000

tgtatcacat attctgctga cgcaccgatg cagccttttt tctcctgcca catgaagcac 3060

ttcacttaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgcaga 3120

tgtccggcgg tgcttttgcc gttacgcact actttagtca gttccgcagt accgtcagta 3180

gctgaacagg agggacagtg ttgatatcgg gtagcaccag aagtctatag catgtgcata 3240

cctttggtcg aaaaaaaaag cccgcactgt caggtgcggg cttttttcag tgtttccttg 3300

ccggattacg ccccgccctg ccactcatcg cagtattgtt gtaattcatt aagcattctg 3360

ccgacatgga agccatcaca aacggcatga tgaacttgga tcgccagtgg cattaacacc 3420

ttgtcgcctt gcgtataata ttttcccata gtgaaaacgg gggcgaagaa gttgtccata 3480

tttgctacgt ttaaatcaaa actggtgaaa ctcacccagg gattggcact gacgaaaaac 3540

atattttcga taaac 3555

<210> 10

<211> 6594

<212> DNA

<213> artificial sequence

<220>

<223> p1884 plasmid

<400> 10

tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag 60

ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc 120

aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt 180

acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg 240

gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa 300

cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa 360

atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt 420

ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt 480

tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc 540

aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg 600

caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt 660

ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat 720

cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat 780

ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat 840

cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc 900

atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa 960

ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact 1020

agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact 1080

tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta 1140

tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga 1200

tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc 1260

gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac 1320

cctcgcaagc tcggttgcgg ccgcaatcgg gcaaatcgct gaatattcct tttgtctccg 1380

accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct 1440

ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa 1500

ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg 1560

ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc 1620

tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc 1680

agcggtatca tcaacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 1740

ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 1800

taaatcaatc taaagtatat atgagtaaac ttggtctgac agttacgttt ccacaaccaa 1860

ttaaccaatt ctgatttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat 1920

atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc 1980

accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc 2040

aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc 2100

accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac 2160

ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt 2220

attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt 2280

acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc 2340

acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt 2400

gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa 2460

ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt 2520

gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc 2580

acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt 2640

ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct 2700

tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg 2760

tgcaatgtaa catcagagat tttgagacac aacgtggctt tccctgcagg atttcggagg 2820

cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2880

gctcgccgca gccgaacgcc gactagtgga ttttacggct agctcagtcc taggtacaat 2940

gctagcgaat tcattaaaga ggagaaaggt acccatggca cgtaccccga gccgtagcag 3000

cattggtagc ctgcgtagtc cgcataccca taaagcaatt ctgaccagca ccattgaaat 3060

cctgaaagaa tgtggttata gcggtctgag cattgaaagc gttgcacgtc gtgccggtgc 3120

aagcaaaccg accatttatc gttggtggac caataaagca gcactgattg ccgaagtgta 3180

tgaaaatgaa agcgaacagg tgcgtaaatt tccggatctg ggtagcttta aagccgatct 3240

ggattttctg ctgcgtaatc tgtggaaagt ttggcgtgaa accatttgtg gtgaagcatt 3300

tcgttgtgtt attgcagaag cacagctgga ccctgcaacc ctgacccagc tgaaagatca 3360

gtttatggaa cgtcgtcgtg agatgccgaa aaaactggtt gaaaatgcca ttagcaatgg 3420

tgaactgccg aaagatacca atcgtgaact gctgctggat atgatttttg gtttttgttg 3480

gtatcgcctg ctgaccgaac agctgaccgt tgaacaggat attgaagaat ttaccttcct 3540

gctaattaat ggtgtttgtc cgggtacaca gcgttaacta gggcccatac ccccaattat 3600

tgaaggccgc taacgcggcc tttttttgtt tctggtctgc ccgacgtacg gtgaatctga 3660

ttcgttacca attgacatga tacgaaacgt accgtatcgt taaggtattt actaactgga 3720

agaggcacta aatgaacacg attaacatcg ctaagaacga cttctctgac atcgaactgg 3780

ctgctatccc gttcaacact ctggctgacc attacggtga gcgtttagct cgcgaacagt 3840

tggcccttga gcatgagtct tacgagatgg gtgaagcacg cttccgcaag atgtttgagc 3900

gtcaacttaa agctggtgag gttgcggata acgctgccgc caagcctctc atcactaccc 3960

tactccctaa gatgattgca cgcatcaacg actggtttga ggaagtgaaa gctaagcgcg 4020

gcaagcgccc gacagccttc cagttcctgc aagaaatcaa gccggaagcc gtagcgtaca 4080

tcaccattaa gaccactctg gcttgcctaa ccagtgctga caatacaacc gttcaggctg 4140

tagcaagcgc aatcggtcgg gccattgagg acgaggctcg cttcggtcgt atccgtgacc 4200

ttgaagctaa gcacttcaag aaaaacgttg aggaacaact caacaagcgc gtagggcacg 4260

tctacaagaa agcatttatg caagttgtcg aggctgacat gctctctaag ggtctactcg 4320

gtggcgaggc gtggtcttcg tggcataagg aagactctat tcatgtagga gtacgctgca 4380

tcgagatgct cattgagtca accggaatgg ttagcttaca ccgccaaaat gctggcgtag 4440

taggtcaaga ctctgagact atcgaactcg cacctgaata cgctgaggct atcgcaaccc 4500

gtgcaggtgc gctggctggc atctctccga tgttccaacc ttgcgtagtt cctcctaagc 4560

cgtggactgg cattactggt ggtggctatt gggctaacgg tcgtcgtcct ctggcgctgg 4620

tgcgtactca cagtaagaaa gcactgatgc gctacgaaga cgtttacatg cctgaggtgt 4680

acaaagcgat taacattgcg caaaacaccg catggaaaat caacaagaaa gtcctagcgg 4740

tcgccaacgt aatcaccaag tggaagcatt gtccggtcga ggacatccct gcgattgagc 4800

gtgaagaact cccgatgaaa ccggaagaca tcgacatgaa tcctgaggct ctcaccgcgt 4860

ggaaacgtgc tgccgctgct gtgtaccgca aggacaaggc tcgcaagtct cgccgtatca 4920

gccttgagtt catgcttgag caagccaata agtttgctaa ccataaggcc atctggttcc 4980

cttacaacat ggactggcgc ggtcgtgttt acgctgtgtc aatgttcaac ccgcaaggta 5040

acgatatgac caaaggactg cttacgctgg cgaaaggtaa accaatcggt aaggaaggtt 5100

actactggct gaaaatccac ggtgcaaact gtgcgggtgt cgataaggtt ccgttccctg 5160

agcgcatcaa gttcattgag gaaaaccacg agaacatcat ggcttgcgct aagtctccac 5220

tggagaacac ttggtgggct gagcaagatt ctccgttctg cttccttgcg ttctgctttg 5280

agtacgctgg ggtacagcac cacggcctga gctataactg ctcccttccg ctggcgtttg 5340

acgggtcttg ctctggcatc cagcacttct ccgcgatgct ccgagatgag gtaggtggtc 5400

gcgcggttaa cttgcttcct agtgaaaccg ttcaggacat ctacgggatt gttgctaaga 5460

aagtcaacga gattctacaa gcagacgcaa tcaatgggac cgataacgaa gtagttaccg 5520

tgaccgatga gaacactggt gaaatctctg agaaagtcaa gctgggcact aaggcactgg 5580

ctggtcaatg gctggcttac ggtgttactc gcagtgtgac taagcgttca gtcatgacgc 5640

tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt gctggaagat accattcagc 5700

cagctattga ttccggcaag ggtctgatgt tcactcagcc gaatcaggct gctggataca 5760

tggctaagct gatttgggaa tctgtgagcg tgacggtggt agctgcggtt gaagcaatga 5820

actggcttaa gtctgctgct aagctgctgg ctgctgaggt caaagataag aagactggag 5880

agattcttcg caagcgttgc gctgtgcatt gggtaactcc tgatggtttc cctgtgtggc 5940

aggaatacaa gaagcctatt cagacgcgct tgaacctgat gttcctcggt cagttccgct 6000

tacagcctac cattaacacc aacaaagata gcgagattga tgcacacaaa caggagtctg 6060

gtatcgctcc taactttgta cacagccaag acggtagcca ccttcgtaag actgtagtgt 6120

gggcacacga gaagtacgga atcgaatctt ttgcactgat tcacgactcc ttcggtacca 6180

ttccggctga cgctgcgaac ctgttcaaag cagtgcgcga aactatggtt gacacatatg 6240

agtcttgtga tgtactggct gatttctacg accagttcgc tgaccagttg cacgagtctc 6300

aattggacaa aatgccagca cttccggcta aaggtaactt gaacctccgt gacatcttag 6360

agtcggactt cgcgttcgcg gcggctaacg acgagaacta cgctgcggca gtgtaataat 6420

gacgcatcct cacgataata tccgggtagg acgaacaata aggccgcaaa tcgcggcctt 6480

ttttattgat aacaaaagga cagttttccc tttgatatgt aacggtgaac agttgttcta 6540

cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacc 6594

<210> 11

<211> 6594

<212> DNA

<213> artificial sequence

<220>

<223> p1885 plasmid

<400> 11

tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag 60

ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc 120

aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt 180

acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg 240

gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa 300

cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa 360

atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt 420

ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt 480

tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc 540

aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg 600

caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt 660

ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat 720

cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat 780

ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat 840

cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc 900

atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa 960

ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact 1020

agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact 1080

tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta 1140

tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga 1200

tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc 1260

gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac 1320

cctcgcaagc tcggttgcgg ccgcaatcgg gcaaatcgct gaatattcct tttgtctccg 1380

accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct 1440

ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa 1500

ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg 1560

ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc 1620

tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc 1680

agcggtatca tcaacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 1740

ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 1800

taaatcaatc taaagtatat atgagtaaac ttggtctgac agttacgttt ccacaaccaa 1860

ttaaccaatt ctgatttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat 1920

atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc 1980

accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc 2040

aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc 2100

accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac 2160

ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt 2220

attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt 2280

acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc 2340

acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt 2400

gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa 2460

ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt 2520

gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc 2580

acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt 2640

ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct 2700

tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg 2760

tgcaatgtaa catcagagat tttgagacac aacgtggctt tccctgcagg atttcggagg 2820

cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2880

gctcgccgca gccgaacgcc gactagtgga ttttacggct agctcagtcc taggtacaat 2940

gctagcgaat tcattaaaga ggagaaaggt acccatggca cgtaccccga gccgtagcag 3000

cattggtagc ctgcgtagtc cgcataccca taaagcaatt ctgaccagca ccattgaaat 3060

cctgaaagaa tgtggttata gcggtctgag cattgaaagc gttgcacgtc gtgccggtgc 3120

aagcaaaccg accatttatc gttggtggac caataaagca gcactgattg ccgaagtgta 3180

tgaaaatgaa agcgaacagg tgcgtaaatt tccggatctg ggtagcttta aagccgatct 3240

ggattttctg ctgcgtaatc tgtggaaagt ttggcgtgaa accatttgtg gtgaagcatt 3300

tcgttgtgtt attgcagaag cacagctgga ccctgcaacc ctgacccagc tgaaagatca 3360

gtttatggaa cgtcgtcgtg agatgccgaa aaaactggtt gaaaatgcca ttagcaatgg 3420

tgaactgccg aaagatacca atcgtgaact gctgctggat atgatttttg gtttttgttg 3480

gtatcgcctg ctgaccgaac agctgaccgt tgaacaggat attgaagaat ttaccttcct 3540

gctaattaat ggtgtttgtc cgggtacaca gcgttaacta gggcccatac ccccaattat 3600

tgaaggccgc taacgcggcc tttttttgtt tctggtctgc ccgacgtacg gtgaatctga 3660

ttcgttacca attgacatga tacgaaacgt accgtatcgt taaggtattt actaactgga 3720

agaggcacta aatgaacacg attaacatcg ctaagaacga cttctctgac atcgaactgg 3780

ctgctatccc gttcaacact ctggctgacc attacggtga gcgtttagct cgcgaacagt 3840

tggcccttga gcatgagtct tacgagatgg gtgaagcacg cttccgcaag atgtttgagc 3900

gtcaacttaa agctggtgag gttgcggata acgctgccgc caagcctctc atcactaccc 3960

tactccctaa gatgattgca cgcatcaacg actggtttga ggaagtgaaa gctaagcgcg 4020

gcaagcgccc gacagccttc cagttcctgc aagaaatcaa gccggaagcc gtagcgtaca 4080

tcaccattaa gaccactctg gcttgcctaa ccagtgctga caatacaacc gttcaggctg 4140

tagcaagcgc aatcggtcgg gccattgagg acgaggctcg cttcggtcgt atccgtgacc 4200

ttgaagctaa gcacttcaag aaaaacgttg aggaacaact caacaagcgc gtagggcacg 4260

tctacaagaa agcatttatg caagttgtcg aggctgacat gctctctaag ggtctactcg 4320

gtggcgaggc gtggtcttcg tggcataagg aagactctat tcatgtagga gtacgctgca 4380

tcgagatgct cattgagtca accggaatgg ttagcttaca ccgccaaaat gctggcgtag 4440

taggtcaaga ctctgagact atcgaactcg cacctgaata cgctgaggct atcgcaaccc 4500

gtgcaggtgc gctggctggc atctctccga tgttccaacc ttgcgtagtt cctcctaagc 4560

cgtggactgg cattactggt ggtggctatt gggctaacgg tcgtcgtcct ctggcgctgg 4620

tgcgtactca cagtaagaaa gcactgatgc gctacgaaga cgtttacatg cctgaggtgt 4680

acaaagcgat taacattgcg caaaacaccg catggaaaat caacaagaaa gtcctagcgg 4740

tcgccaacgt aatcaccaag tggaagcatt gtccggtcga ggacatccct gcgattgagc 4800

gtgaagaact cccgatgaaa ccggaagaca tcgacatgaa tcctgaggct ctcaccgcgt 4860

ggaaacgtgc tgccgctgct gtgtaccgca aggacaaggc tcgcaagtct cgccgtatca 4920

gccttgagtt catgcttgag caagccaata agtttgctaa ccataaggcc atctggttcc 4980

cttacaacat ggactggcgc ggtcgtgttt acgctgtgtc aatgttcaac ccgcaaggta 5040

acgatatgac caaaggactg cttacgctgg cgaaaggtaa accaatcggt aaggaaggtt 5100

actactggct gaaaatccac ggtgcaaact gtgcgggtgt cgataaggtt ccgttccctg 5160

agcgcatcaa gttcattgag gaaaaccacg agaacatcat ggcttgcgct aagtctccac 5220

tggagaacac ttggtgggct gagcaagatt ctccgttctg cttccttgcg ttctgctttg 5280

agtacgctgg ggtacagcac cacggcctga gctataactg ctcccttccg ctggcgtttg 5340

acgggtcttg ctctggcatc cagcacttct ccgcgatgct ccgagatgag gtaggtggtc 5400

gcgcggttaa cttgcttcct agtgaaaccg ttcaggacat ctacgggatt gttgctaaga 5460

aagtcaacga gattctacaa gcagacgcaa tcaatgggac cgataacgaa gtagttaccg 5520

tgaccgatga gaacactggt gaaatctctg agaaagtcaa gctgggcact aaggcactgg 5580

ctggtcaatg gctggcttac ggtgttactc gcagtgtgac taagcgttca gtcatgacgc 5640

tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt gctggaagat accattcagc 5700

cagctattga ttccggcaag ggtctgatgt tcactcagcc gaatcaggct gctggataca 5760

tggctaagct gatttgggaa tctgtgagcg tgacggtggt agctgcggtt gaagcaatga 5820

actggcttaa gtctgctgct aagctgctgg ctgctgaggt caaagataag aagactggag 5880

agattcttcg caagcgttgc gctgtgcatt gggtaactcc tgatggtttc cctgtgtggc 5940

aggaatacaa gaagcctatt cagacgcgct tgaacctgat gttcctcggt cagttccgct 6000

tacagcctac cattaacacc aacaaagata gcgagattga tgcacacaaa caggagtctg 6060

gtatcgctcc taactttgta cacagccaag acggtagcca ccttcgtaag actgtagtgt 6120

gggcacacga gaagtacgga atcgaatctt ttgcactgat tcacgactcc ttcggtacca 6180

ttccggctga cgctgcgaac ctgttcaaag cagtgcgcga aactatggtt gacacatatg 6240

agtcttgtga tgtactggct gatttctacg accagttcgc tgaccagttg cacgagtctc 6300

aattggacaa aatgccagca cttccggcta aaggtaactt gaacctccgt gacatcttag 6360

agtcggactt cgcgttcgcg gcagcgaacg acgaaaacta tgccctggta gcctaataat 6420

gacgcatcct cacgataata tccgggtagg acgaacaata aggccgcaaa tcgcggcctt 6480

ttttattgat aacaaaagga cagttttccc tttgatatgt aacggtgaac agttgttcta 6540

cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacc 6594

<210> 12

<211> 2682

<212> DNA

<213> artificial sequence

<220>

<223> T7 RNA polymerase version AAV

<400> 12

atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg 60

ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag 120

catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa 180

gctggtgagg ttgcggataa cgctgccgcc aagcctctca tcactaccct actccctaag 240

atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg 300

acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag 360

accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca 420

atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag 480

cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa 540

gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg 600

tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc 660

attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac 720

tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg 780

ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc 840

attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac 900

agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt 960

aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta 1020

atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc 1080

ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct 1140

gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc 1200

atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg 1260

gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc 1320

aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg 1380

aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag 1440

ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact 1500

tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg 1560

gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc 1620

tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac 1680

ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag 1740

attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag 1800

aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg 1860

ctggcttacg gtgttactcg cagtgtgact aagcgttcag tcatgacgct ggcttacggg 1920

tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat 1980

tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg 2040

atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag 2100

tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc 2160

aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag 2220

aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc 2280

attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct 2340

aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag 2400

aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac 2460

gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat 2520

gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa 2580

atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc 2640

gcgttcgcgg cggctaacga cgagaactac gctgcggcag tg 2682

<210> 13

<211> 894

<212> PRT

<213> artificial sequence

<220>

<223> T7 RNA polymerase version AAV

<400> 13

Met Asn Thr Ile Asn Ile Ala Lys Asn Asp Phe Ser Asp Ile Glu Leu

1 5 10 15

Ala Ala Ile Pro Phe Asn Thr Leu Ala Asp His Tyr Gly Glu Arg Leu

20 25 30

Ala Arg Glu Gln Leu Ala Leu Glu His Glu Ser Tyr Glu Met Gly Glu

35 40 45

Ala Arg Phe Arg Lys Met Phe Glu Arg Gln Leu Lys Ala Gly Glu Val

50 55 60

Ala Asp Asn Ala Ala Ala Lys Pro Leu Ile Thr Thr Leu Leu Pro Lys

65 70 75 80

Met Ile Ala Arg Ile Asn Asp Trp Phe Glu Glu Val Lys Ala Lys Arg

85 90 95

Gly Lys Arg Pro Thr Ala Phe Gln Phe Leu Gln Glu Ile Lys Pro Glu

100 105 110

Ala Val Ala Tyr Ile Thr Ile Lys Thr Thr Leu Ala Cys Leu Thr Ser

115 120 125

Ala Asp Asn Thr Thr Val Gln Ala Val Ala Ser Ala Ile Gly Arg Ala

130 135 140

Ile Glu Asp Glu Ala Arg Phe Gly Arg Ile Arg Asp Leu Glu Ala Lys

145 150 155 160

His Phe Lys Lys Asn Val Glu Glu Gln Leu Asn Lys Arg Val Gly His

165 170 175

Val Tyr Lys Lys Ala Phe Met Gln Val Val Glu Ala Asp Met Leu Ser

180 185 190

Lys Gly Leu Leu Gly Gly Glu Ala Trp Ser Ser Trp His Lys Glu Asp

195 200 205

Ser Ile His Val Gly Val Arg Cys Ile Glu Met Leu Ile Glu Ser Thr

210 215 220

Gly Met Val Ser Leu His Arg Gln Asn Ala Gly Val Val Gly Gln Asp

225 230 235 240

Ser Glu Thr Ile Glu Leu Ala Pro Glu Tyr Ala Glu Ala Ile Ala Thr

245 250 255

Arg Ala Gly Ala Leu Ala Gly Ile Ser Pro Met Phe Gln Pro Cys Val

260 265 270

Val Pro Pro Lys Pro Trp Thr Gly Ile Thr Gly Gly Gly Tyr Trp Ala

275 280 285

Asn Gly Arg Arg Pro Leu Ala Leu Val Arg Thr His Ser Lys Lys Ala

290 295 300

Leu Met Arg Tyr Glu Asp Val Tyr Met Pro Glu Val Tyr Lys Ala Ile

305 310 315 320

Asn Ile Ala Gln Asn Thr Ala Trp Lys Ile Asn Lys Lys Val Leu Ala

325 330 335

Val Ala Asn Val Ile Thr Lys Trp Lys His Cys Pro Val Glu Asp Ile

340 345 350

Pro Ala Ile Glu Arg Glu Glu Leu Pro Met Lys Pro Glu Asp Ile Asp

355 360 365

Met Asn Pro Glu Ala Leu Thr Ala Trp Lys Arg Ala Ala Ala Ala Val

370 375 380

Tyr Arg Lys Asp Lys Ala Arg Lys Ser Arg Arg Ile Ser Leu Glu Phe

385 390 395 400

Met Leu Glu Gln Ala Asn Lys Phe Ala Asn His Lys Ala Ile Trp Phe

405 410 415

Pro Tyr Asn Met Asp Trp Arg Gly Arg Val Tyr Ala Val Ser Met Phe

420 425 430

Asn Pro Gln Gly Asn Asp Met Thr Lys Gly Leu Leu Thr Leu Ala Lys

435 440 445

Gly Lys Pro Ile Gly Lys Glu Gly Tyr Tyr Trp Leu Lys Ile His Gly

450 455 460

Ala Asn Cys Ala Gly Val Asp Lys Val Pro Phe Pro Glu Arg Ile Lys

465 470 475 480

Phe Ile Glu Glu Asn His Glu Asn Ile Met Ala Cys Ala Lys Ser Pro

485 490 495

Leu Glu Asn Thr Trp Trp Ala Glu Gln Asp Ser Pro Phe Cys Phe Leu

500 505 510

Ala Phe Cys Phe Glu Tyr Ala Gly Val Gln His His Gly Leu Ser Tyr

515 520 525

Asn Cys Ser Leu Pro Leu Ala Phe Asp Gly Ser Cys Ser Gly Ile Gln

530 535 540

His Phe Ser Ala Met Leu Arg Asp Glu Val Gly Gly Arg Ala Val Asn

545 550 555 560

Leu Leu Pro Ser Glu Thr Val Gln Asp Ile Tyr Gly Ile Val Ala Lys

565 570 575

Lys Val Asn Glu Ile Leu Gln Ala Asp Ala Ile Asn Gly Thr Asp Asn

580 585 590

Glu Val Val Thr Val Thr Asp Glu Asn Thr Gly Glu Ile Ser Glu Lys

595 600 605

Val Lys Leu Gly Thr Lys Ala Leu Ala Gly Gln Trp Leu Ala Tyr Gly

610 615 620

Val Thr Arg Ser Val Thr Lys Arg Ser Val Met Thr Leu Ala Tyr Gly

625 630 635 640

Ser Lys Glu Phe Gly Phe Arg Gln Gln Val Leu Glu Asp Thr Ile Gln

645 650 655

Pro Ala Ile Asp Ser Gly Lys Gly Leu Met Phe Thr Gln Pro Asn Gln

660 665 670

Ala Ala Gly Tyr Met Ala Lys Leu Ile Trp Glu Ser Val Ser Val Thr

675 680 685

Val Val Ala Ala Val Glu Ala Met Asn Trp Leu Lys Ser Ala Ala Lys

690 695 700

Leu Leu Ala Ala Glu Val Lys Asp Lys Lys Thr Gly Glu Ile Leu Arg

705 710 715 720

Lys Arg Cys Ala Val His Trp Val Thr Pro Asp Gly Phe Pro Val Trp

725 730 735

Gln Glu Tyr Lys Lys Pro Ile Gln Thr Arg Leu Asn Leu Met Phe Leu

740 745 750

Gly Gln Phe Arg Leu Gln Pro Thr Ile Asn Thr Asn Lys Asp Ser Glu

755 760 765

Ile Asp Ala His Lys Gln Glu Ser Gly Ile Ala Pro Asn Phe Val His

770 775 780

Ser Gln Asp Gly Ser His Leu Arg Lys Thr Val Val Trp Ala His Glu

785 790 795 800

Lys Tyr Gly Ile Glu Ser Phe Ala Leu Ile His Asp Ser Phe Gly Thr

805 810 815

Ile Pro Ala Asp Ala Ala Asn Leu Phe Lys Ala Val Arg Glu Thr Met

820 825 830

Val Asp Thr Tyr Glu Ser Cys Asp Val Leu Ala Asp Phe Tyr Asp Gln

835 840 845

Phe Ala Asp Gln Leu His Glu Ser Gln Leu Asp Lys Met Pro Ala Leu

850 855 860

Pro Ala Lys Gly Asn Leu Asn Leu Arg Asp Ile Leu Glu Ser Asp Phe

865 870 875 880

Ala Phe Ala Ala Ala Asn Asp Glu Asn Tyr Ala Ala Ala Val

885 890

<210> 14

<211> 2682

<212> DNA

<213> artificial sequence

<220>

<223> T7 RNA polymerase version LVA

<400> 14

atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg 60

ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag 120

catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa 180

gctggtgagg ttgcggataa cgctgccgcc aagcctctca tcactaccct actccctaag 240

atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg 300

acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag 360

accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca 420

atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag 480

cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa 540

gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg 600

tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc 660

attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac 720

tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg 780

ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc 840

attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac 900

agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt 960

aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta 1020

atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc 1080

ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct 1140

gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc 1200

atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg 1260

gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc 1320

aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg 1380

aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag 1440

ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact 1500

tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg 1560

gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc 1620

tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac 1680

ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag 1740

attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag 1800

aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg 1860

ctggcttacg gtgttactcg cagtgtgact aagcgttcag tcatgacgct ggcttacggg 1920

tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat 1980

tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg 2040

atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag 2100

tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc 2160

aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag 2220

aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc 2280

attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct 2340

aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag 2400

aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac 2460

gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat 2520

gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa 2580

atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc 2640

gcgttcgcgg cagcgaacga cgaaaactat gccctggtag cc 2682

<210> 15

<211> 894

<212> PRT

<213> artificial sequence

<220>

<223> T7 RNA polymerase version LVA

<400> 15

Met Asn Thr Ile Asn Ile Ala Lys Asn Asp Phe Ser Asp Ile Glu Leu

1 5 10 15

Ala Ala Ile Pro Phe Asn Thr Leu Ala Asp His Tyr Gly Glu Arg Leu

20 25 30

Ala Arg Glu Gln Leu Ala Leu Glu His Glu Ser Tyr Glu Met Gly Glu

35 40 45

Ala Arg Phe Arg Lys Met Phe Glu Arg Gln Leu Lys Ala Gly Glu Val

50 55 60

Ala Asp Asn Ala Ala Ala Lys Pro Leu Ile Thr Thr Leu Leu Pro Lys

65 70 75 80

Met Ile Ala Arg Ile Asn Asp Trp Phe Glu Glu Val Lys Ala Lys Arg

85 90 95

Gly Lys Arg Pro Thr Ala Phe Gln Phe Leu Gln Glu Ile Lys Pro Glu

100 105 110

Ala Val Ala Tyr Ile Thr Ile Lys Thr Thr Leu Ala Cys Leu Thr Ser

115 120 125

Ala Asp Asn Thr Thr Val Gln Ala Val Ala Ser Ala Ile Gly Arg Ala

130 135 140

Ile Glu Asp Glu Ala Arg Phe Gly Arg Ile Arg Asp Leu Glu Ala Lys

145 150 155 160

His Phe Lys Lys Asn Val Glu Glu Gln Leu Asn Lys Arg Val Gly His

165 170 175

Val Tyr Lys Lys Ala Phe Met Gln Val Val Glu Ala Asp Met Leu Ser

180 185 190

Lys Gly Leu Leu Gly Gly Glu Ala Trp Ser Ser Trp His Lys Glu Asp

195 200 205

Ser Ile His Val Gly Val Arg Cys Ile Glu Met Leu Ile Glu Ser Thr

210 215 220

Gly Met Val Ser Leu His Arg Gln Asn Ala Gly Val Val Gly Gln Asp

225 230 235 240

Ser Glu Thr Ile Glu Leu Ala Pro Glu Tyr Ala Glu Ala Ile Ala Thr

245 250 255

Arg Ala Gly Ala Leu Ala Gly Ile Ser Pro Met Phe Gln Pro Cys Val

260 265 270

Val Pro Pro Lys Pro Trp Thr Gly Ile Thr Gly Gly Gly Tyr Trp Ala

275 280 285

Asn Gly Arg Arg Pro Leu Ala Leu Val Arg Thr His Ser Lys Lys Ala

290 295 300

Leu Met Arg Tyr Glu Asp Val Tyr Met Pro Glu Val Tyr Lys Ala Ile

305 310 315 320

Asn Ile Ala Gln Asn Thr Ala Trp Lys Ile Asn Lys Lys Val Leu Ala

325 330 335

Val Ala Asn Val Ile Thr Lys Trp Lys His Cys Pro Val Glu Asp Ile

340 345 350

Pro Ala Ile Glu Arg Glu Glu Leu Pro Met Lys Pro Glu Asp Ile Asp

355 360 365

Met Asn Pro Glu Ala Leu Thr Ala Trp Lys Arg Ala Ala Ala Ala Val

370 375 380

Tyr Arg Lys Asp Lys Ala Arg Lys Ser Arg Arg Ile Ser Leu Glu Phe

385 390 395 400

Met Leu Glu Gln Ala Asn Lys Phe Ala Asn His Lys Ala Ile Trp Phe

405 410 415

Pro Tyr Asn Met Asp Trp Arg Gly Arg Val Tyr Ala Val Ser Met Phe

420 425 430

Asn Pro Gln Gly Asn Asp Met Thr Lys Gly Leu Leu Thr Leu Ala Lys

435 440 445

Gly Lys Pro Ile Gly Lys Glu Gly Tyr Tyr Trp Leu Lys Ile His Gly

450 455 460

Ala Asn Cys Ala Gly Val Asp Lys Val Pro Phe Pro Glu Arg Ile Lys

465 470 475 480

Phe Ile Glu Glu Asn His Glu Asn Ile Met Ala Cys Ala Lys Ser Pro

485 490 495

Leu Glu Asn Thr Trp Trp Ala Glu Gln Asp Ser Pro Phe Cys Phe Leu

500 505 510

Ala Phe Cys Phe Glu Tyr Ala Gly Val Gln His His Gly Leu Ser Tyr

515 520 525

Asn Cys Ser Leu Pro Leu Ala Phe Asp Gly Ser Cys Ser Gly Ile Gln

530 535 540

His Phe Ser Ala Met Leu Arg Asp Glu Val Gly Gly Arg Ala Val Asn

545 550 555 560

Leu Leu Pro Ser Glu Thr Val Gln Asp Ile Tyr Gly Ile Val Ala Lys

565 570 575

Lys Val Asn Glu Ile Leu Gln Ala Asp Ala Ile Asn Gly Thr Asp Asn

580 585 590

Glu Val Val Thr Val Thr Asp Glu Asn Thr Gly Glu Ile Ser Glu Lys

595 600 605

Val Lys Leu Gly Thr Lys Ala Leu Ala Gly Gln Trp Leu Ala Tyr Gly

610 615 620

Val Thr Arg Ser Val Thr Lys Arg Ser Val Met Thr Leu Ala Tyr Gly

625 630 635 640

Ser Lys Glu Phe Gly Phe Arg Gln Gln Val Leu Glu Asp Thr Ile Gln

645 650 655

Pro Ala Ile Asp Ser Gly Lys Gly Leu Met Phe Thr Gln Pro Asn Gln

660 665 670

Ala Ala Gly Tyr Met Ala Lys Leu Ile Trp Glu Ser Val Ser Val Thr

675 680 685

Val Val Ala Ala Val Glu Ala Met Asn Trp Leu Lys Ser Ala Ala Lys

690 695 700

Leu Leu Ala Ala Glu Val Lys Asp Lys Lys Thr Gly Glu Ile Leu Arg

705 710 715 720

Lys Arg Cys Ala Val His Trp Val Thr Pro Asp Gly Phe Pro Val Trp

725 730 735

Gln Glu Tyr Lys Lys Pro Ile Gln Thr Arg Leu Asn Leu Met Phe Leu

740 745 750

Gly Gln Phe Arg Leu Gln Pro Thr Ile Asn Thr Asn Lys Asp Ser Glu

755 760 765

Ile Asp Ala His Lys Gln Glu Ser Gly Ile Ala Pro Asn Phe Val His

770 775 780

Ser Gln Asp Gly Ser His Leu Arg Lys Thr Val Val Trp Ala His Glu

785 790 795 800

Lys Tyr Gly Ile Glu Ser Phe Ala Leu Ile His Asp Ser Phe Gly Thr

805 810 815

Ile Pro Ala Asp Ala Ala Asn Leu Phe Lys Ala Val Arg Glu Thr Met

820 825 830

Val Asp Thr Tyr Glu Ser Cys Asp Val Leu Ala Asp Phe Tyr Asp Gln

835 840 845

Phe Ala Asp Gln Leu His Glu Ser Gln Leu Asp Lys Met Pro Ala Leu

850 855 860

Pro Ala Lys Gly Asn Leu Asn Leu Arg Asp Ile Leu Glu Ser Asp Phe

865 870 875 880

Ala Phe Ala Ala Ala Asn Asp Glu Asn Tyr Ala Leu Val Ala

885 890

<210> 16

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1334 primers

<400> 16

ggacctccca ccattccaag 20

<210> 17

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1335 primer

<400> 17

acggcgatgt tcaggttctt 20

<210> 18

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1336 primer

<400> 18

ggcgaaagaa gacctggtca 20

<210> 19

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1337 primers

<400> 19

tagccggcga aatggatgtt 20

<210> 20

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1322 primer

<400> 20

catcagaccg cattcgcttg 20

<210> 21

<211> 20

<212> DNA

<213> artificial sequence

<220>

<223> AD1323 primer

<400> 21

ggacgaagat gtggaagcca 20

<210> 22

<211> 32767

<212> DNA

<213> BW4 phage

<400> 22

agcgatatct ccccgggttt ttccacaggg tgtccgccca gggcgtcgct gtcgagctca 60

cagcgcacgc tgaacgcccg ccagcactcg aggcgatccg aacagcctcg ctccgagtgc 120

gtcgggcctg tgtggatcgc tcatgagttt cgtaacaagc ccctagccac agcccgattc 180

agatagaata ggagcatgga agggcagtgc ggatggtgcg gtcgggcatt cgatcgtgcc 240

cggacgggtc gcccgcgacg cttctgctcg gcccgctgtc gggtcgccgc gtcccggtgt 300

gcgatcccgc tggccatgag gtcccgcact gcgtgggtcc gctgcgacgg caagcgcccc 360

atcaccctgg ctggcgctcc ggcctcatcc acggacccgg gcacatggtc tggctggtcg 420

caggtgcgac gcgccacggc cggcgatggc ttcgggacca tgctcggtga cgggctgggg 480

tgctgggatc tcgaccactt cgacgatcag ggcgcccggg ccttcatcga ccggatcgat 540

aagccgatca tcttcgccga gcggtcggtg tcggggcatg gcttccacat cttcgtccgg 600

actgacgagg cccccggacg ccgcaccgga aacatcgagt tctactcacg ccatcggttc 660

atcagggtca caggagacca gttcgtctga agaagggggt gcgccatggc tgcacaggtc 720

agggccgtgg accccgatga gcgcccaccc gcccgcaagc gggccaagac catcacccag 780

gccgcgaagt ccggcactga ggttgaactg ttggaggcac tgcaggctcg cgtggcccgc 840

gccgtgcagg accgtgacac tccgccgcgc gatctggcag cgctgacgaa gcggctgatg 900

gacatcaccc gggagctcga ggcggcccgg gtcaaggatc aggaggcggg atctgatggt 960

gccgtcaccg cagacgaaac atggcgaccg caagctctct gaggtcgcca agcacctgat 1020

ccttcctgaa gggatcgtct cgacgggctg gccggccgtg cgtgaccggt gtggcgagtg 1080

gggtgtggtc ttcgaccgtt ggcaggacgg catgggccgg gtgatcctgt cgaagcgcgg 1140

cagcggcctg ttcgccgctg gtgtgggcgg ggtcggcatg tcgatcccgc gccagaccgg 1200

caagaccttc accgtcggca tgatcatcct cgggctgtgc tcgctgagcg aggagctcac 1260

ggtgctgtgg acctcccacc attccaagac gaccaccaag actttcgagt cgctgcgggg 1320

catggcccag cgtaagaagg tcgccccgtt gatccgtcag gtccgaacag gaaacggtga 1380

ccagcagatc attttcagca acggttcgag gatctacttc ggtgcccggg aacagggctt 1440

cgggcgtggc ttcgacgacg tggacatcga gatctttgac gaggcgcaga tcctgtccga 1500

gcaggccctc tccgacatgg ttcccgcggc gaatgtgagc accaatccgc tgatcatctt 1560

catgggcacc ccgccgcgtc cctcggaccc gtcggaggcg ttcgcgaacc gccgcgccga 1620

agctctggcg ggcgacgccc cggacgccgc ctggatcgaa ttcggagcgg acgagcacgc 1680

cgacccgacc agccgcgccc aatggcgtaa ggcaaaccca tcctttcctc accgcacgtc 1740

ggagacctcc attctgcgga tgaagaagat gctcgggccc gagtccttca aacgcgaggg 1800

cttgggcatc tgggatgaga cggcatcggt ccgcgcgatc ccagccgaag ggtggcgcgt 1860

cctgaccgtc aaggaaccac ccgccgacgc gatccagtcc ttcggcatca agttcgccat 1920

cgacgggagt gcggtcgccc tggcagccgc cctgaaaccc aaggacgggc cgatctatgt 1980

cgaaggaatc gagcagcgct cggcatccga cggcatcgaa tggctcgccg actacctgac 2040

gcccctgtgg cgcaacacgg cccagatcgt catcgatggc aagtccggcg ccggtgccct 2100

ggttgatgcg ctgcgccgtg gtggcgtggc tgcgaaggtg atcctcaccc cgagcgtcgc 2160

cgacgtgatc accgcccaca gcctgactct ggaggccatc aagaccggtg gactgtcgca 2220

cctggctgac ccggagctgg atcggcaggt ccgcatcgcc acgaagcgaa agatcggggc 2280

cgccgggggc ttcggctggc aggcccccga aggcgacacc gtcgccctcc tcgacgccat 2340

cacgcttgcc cactgggcgg ccctcaccac gaagcgacat cccggcagga aggcggtggc 2400

actggcatga gcctcctcgt caacccctat gcgtcgccgt ccttcttctc gtccccgtcc 2460

gtggtcggac tcggagcaga cgagcaggag ctcctggacg agctggtggc cctgtgggca 2520

cgcaagaagc cccgcaacgt gctgcgcggc ctgtaccttg acggcaagca gcagatcaag 2580

aacctgaaca tcgccgtgcc cgacgagatc gccgacagtc tccagatcgt ggtcggctgg 2640

cccgagaagg ccgtcttcgg gctatcgaac ctgtgcatgt gggatggcgt cgtcactccc 2700

acaggcgacg agaatccctt cgggcttgac gatctcctgt cggccaaccg cttcgacgtc 2760

gagatcaatg aaacgatcac ctcggccatg gcgaactccg tggccttcct gaccgtatcg 2820

gcgggcaacg tgtccatagg tgagccgccg gtggtgatca tgccgttctc cgccgaatgg 2880

gcctcagccc tgtgggaccg gcgcacccgc tcaatcaagg cgggactgac catcggcgac 2940

atcgactacc tgggccgccc caccagcctc tcgctcttca cccgcaccgc caccatcacc 3000

tgcgtggggt cccggctggg atggatgatc gaagatcgcg ccgagcacgg gctgaaccgc 3060

gtcccgatgg agccggtccc gttccgccca acccttgacc gccccttcgg gcgctcgcgg 3120

atctcgcgcc aggtgatgac catcgtggac cgcgccatgc gcgcggccct gcgcatggac 3180

atctcctcag agctgttcac cgcacccggc ctgctcctca acggaatcac cccggagcag 3240

tgggcagaga tccagaagtg gacatggaag ctcggcacgg tgcgcggcct gactcgcgac 3300

gaggatggcg agaccgcatc ggtcgagacg atcccccagc agtcgatgga accgttcatc 3360

gcgcagctgc gcgagctggc cgaggaattc gcctcagcca catccatgcc gctgtctgca 3420

ttgggggtcg tccaagacaa cccctcctcg gctgacgcca tctacgcggc gaaagaagac 3480

ctggtcatcg aggccaccaa cgccaaccgg atcaccggct acgcgctatc ccgggtcttc 3540

caagacgcgg tgatgatgcg cgacggcctg accgagatgc ccgacgagct cggcggggtc 3600

gccgccaagt ggcgcaaccc ggcgatgccg tcgatcgtgt cccagtccga cgcgatggtc 3660

aagcagattt cggcgatccc cgggctggcc gctaccgacg tcgccttcga acagctcggc 3720

tattcggcgg ctgacatcgt gcggattcgt acccagatgc gccgagccca ggctgcggac 3780

ggcctgactt cgttgctggc caaaccagcc acgtcgtcaa cgcctggcgc ggagccctct 3840

cagtccgcaa gtccgacgga gccagctgca agcactccgc tgccggacct cgaaggggcc 3900

cctggtgacc gatcgtgatg acctgaacca tttccacgag gccaatgacg cgatccagcg 3960

gcgcgcaatc aacgacctga acaagttttg ggcgcggctt gccaagtcag acccgaaagc 4020

cgttcgcgca gccatggact tattcgtccc ccagctcatc gcctcctacg gagagttggc 4080

cgccgaagcc gctgcccgtt ggtatgagga actacggccc gccgacaaga agaacttcca 4140

ggccgaactc gcggaccctg tgtccgacga catcatcgag gcagatgtgg ctgaggccct 4200

ggggaccagc ggcgcctggg acaccgaggc ggtgcgaggg agcctggccg atgcgatcag 4260

gcgtcagatc ttctacatgg cgcgggcgac tgtcgcacgc aacatcgctc acgacccgaa 4320

gcgtccaagg tttgcacgag ttcctcgggg cgcggtcacg tgcgcgttct gcaccatgct 4380

cgcctccagg gggtgggtgt actacaccgc gaagactgcc gggatcacac gaccctggca 4440

tcgcaagtgc gactgccaga tcgtgcctga gtggaaacgc ggcaacatcc atttcgccgg 4500

ctacgaccct gacaagatgt tcgagcagta tgccgaatcg gtcgatgcgg tggggtcgag 4560

cttcgacacg aaggcaatcc tcgccgacat gcgccgacgc catcccgaag cgctgaccga 4620

cggggtcgtc aacatgagtg aaggacaggg tccggtgacc agtgattaga cagtcggtga 4680

acggatgact acgccgtcgg cgctgcgccg ccgcctggaa tggctactgg agaaccgtga 4740

acggcttctc aggagccatg gcgagtcgga ctttgccgag atgctggatg gcgcccgtca 4800

cgagcttgat gaggcccgcg agcaggcagg cctggccgcg cagtcaaacc caatctgtag 4860

caagccccgt tccaccttcg ggtgggcggg gctttgtcat gcccgcatcc gggcatccaa 4920

ttccgtccca ccgcgagggt ggggcgtcga cctggtggcg cgatgccgcc gaactaatcc 4980

ctggaagggg aaactgctat gcacaagaag ctcatgccgt gggtccgtct catcgaggcg 5040

gtcgagactc ctgctggagc cgcccccacg cccgcgatcg atccgaagga tccggcagcc 5100

aatcccacca ctgagccgaa gccggccgac gcgacgtcgg agaagcctct cggcgaggcg 5160

ggcaaggttg cgttggatcg cgagcgcgag gctcgccgca gcgccgacaa gcgcgccagt 5220

gagttggagg cccgtgtgca ccagctcgag gacgcgggca agaccgaggc ccagaagcag 5280

gccgacgaac tcaagcgcac ccagtccgag ctggagacgc tgaggggcga gaaggcacgg 5340

ctggaggtgg cgtccgcgac gggcgtcccg gtcgatctgc tcgctggccc cggcgacgat 5400

ctggatgcct acgcgcaggc cctgaacgcc tggcgcgaca agcagtccga aaagccagcc 5460

gcccctgcgg tggacacccc ttccccttcg ccgtccgggg tgaccggaca gcccgtgcag 5520

ccgaaccgga cggtcgatga actcatcgcg gccgccgaga agaacggcga tctggcaacc 5580

gcgaagcaac tcaaattgat gaagctcgac gcactgcgtc ggacgtcctg atcagaaagg 5640

caccactatg ccgggcatta ccggacaggg caccacctac aaccttccga actatgtggg 5700

ggagcttttt gcggcatctc ccgaagacac cccgctgctg tcggcgatcg ggggactgac 5760

cggcggcgag tcggtcggcg cccgccagtt cgaatggcag ggctacgacc tgcgcgacgc 5820

cgacggttcg cgccagcgcc tcgagggagc caacgccccc gacggtgagg agcgcacccg 5880

ctactccgcc tccaatgtgg tcgagatcca ccaggagtcg gtggaggtgt cctacaccaa 5940

gcaggccgcg aaccgtgagc gggctaccaa cggtgccgcc acggtccagc tggcgggctc 6000

cgtgctgccg gccgatgagc tcacctggca gatcgaccag cagctcaagc aggtcgcccg 6060

cgatgtcgag aagtccttca tcgcgggcac ctaccagctg cccaccgaca acgccaagcc 6120

gcgccgcacg cgtggcctgc tggaggcgac caccacgaac gtggccgcct cgacccacac 6180

cgcaaaggaa ctcaccgtgg aggagatcct cgacctgttc cagaaggtgt gggagaacgg 6240

cggcatccag gaagccgaga cccgcaccgt cattgtcggt gccgccctga agcggaccct 6300

gacgcgcctg ttcatcaccg acgtcaagta ccaggaagaa tcccgcaacg ttggcggtgt 6360

gaacctgcag accttcgaaa ccgacttcgg caaggcgaac atcatgctcg accgcttcat 6420

gccgagcgac accctcgtgg tcgcgtcgct ggaggacctg aagccggcct tcctcgacat 6480

ccccggcaag ggccacttct tcgccgagcc gctcgccaag accggtgcag ccgacaaggt 6540

gcagatttac ggcgaggtcg ggctgcagta cgggaaccag cgcaagcacg gaaagctcac 6600

tgtcgcaccc gcaacccccg ccaagtaatc acggatcggt ttgaggttgc ctgatgaaag 6660

tcacctcgac catcccgaac ctgactgttc tcgacctgga catccagttc gttgacggtc 6720

aggccgatgt ggacccgcat ctcgccgaga ggctgcgtcg cctcgagcct ctcggcgtgc 6780

gggtccccac agccagccgc aagccgccca cgcggtcgcg gcgtaagcag ggggtcagcc 6840

atggtcgcac ctgatccgga actgccgttc gccaccgtct ccgatatgga gagccggtgg 6900

cgttctttgt ctaaggacga gcacacgcgg gccgaggccc ttctggacga tgcgagcggg 6960

ttgatcgttg atacctgccc gcgctgggaa caggcctcac cggccaccct gcggcgtgtg 7020

acgtgctctg tcgtgcgccg ggcgatggcc gcagacgatg aggacatcgg cgcaacctcg 7080

ctcatggaca cgacgggccc cttcaccact cagcgcgcct actcatcacc ggccggggat 7140

ctcttcttga ccaaggccga gaaggccgcg ctcggcgggg tcaccggcgc attcgagacg 7200

agccttctgg ggctgacatg aagcgctcat ggccgacacc cgtggaacgt ctccgcgagg 7260

gtccgcccga gattgaccgt gacggtgatc cgattgccgg ctccggagtg atcaccaagg 7320

atcctctccc tgatgccctg ttcgcgccgg gcggctcgca gatcctcgtc gcccccggcg 7380

tggcggcagt cgtggacgaa cccaccctct actggcgcgg atcagaagtg atcgatgtgg 7440

tggccaccga caaggtccgg atagccggcc gagtctggac ccctgaagga aatcctgcgc 7500

gatggccgaa gggcgtcgtg ctcaagctca aggcccagga ggcaaagaat cgtggctaat 7560

ttccgtttcg aacccaatac gaaggcgttc accgagtggg cgcagcgcga ctgcgacgcg 7620

cacctgatcg ccggcatcac ggcctcgatg ggggccaagg cgggcgaggg tttctcgacg 7680

atggtctcca acaatggcga ccgcacccgc ggttatctcg cgacggcctc cacgaagggc 7740

cgtatgcggc aggcgcaggg gcatgtcatc gagcgggtca tcggatcgag cggcgtgtga 7800

aaccgcccga cctccacacg ctcgtcgccc accatctggc tgagctcctc gacgtgccgg 7860

tcgtctccac ccgccccgag ggagagacgg cgccgtccaa gttcgttcgg atcatctcga 7920

ccggcggagc gggccgctat ggccgggtct tccagggcat ccagctgacg atcggctcct 7980

acgcgggatc ggcggcgacc gcccgtgatc tcgcgatgca ggtggacgag gccatgaatg 8040

ggctgccggt ctcgccgttg ccggtctcca aggtcaccgg caacaccccg tcggacgacc 8100

ccgatcccga cactcagcag gcccgccaca cggccaccta ccaactcacc acccttatct 8160

cttaggagtc attcatggct gtcaattccg tcaacgtgca cgtcttcggg tccgatgacg 8220

acgtgctcta cctgggcccg tcaggtctga atctgggcaa catttcgctg gaaaccgcga 8280

tcccgaagga gatgatcgac accggctggc tcactgatga cggtgtgacc ctcggcatga 8340

aggactctgt caaggccatc cagggccacc agggccacgc gaatgtgctt cagttcatgg 8400

actcgtcgga taccaccctc gaggcgaccc tcatggagtc tcagctgcag accttcctgt 8460

ggaacctcga cgcggacgct gaggacatcg acggggtcac caagatcacc gcggccagct 8520

cccgcaaggt cctcaacctg tgcgcgatct gggacacctt cgacacccag cacagcggca 8580

tccattggcg ctacgtcttc ccctcgctca ccctgggcga gcgcgatgac atccccttca 8640

aggtgggcga agccagcgct tacaagtatt cgctgggtgt gctggagaag ttcttcgtct 8700

tcaccaacgc ggcagcgatg aaggccggtg gagcatccgc caagacggtg accggtgtga 8760

agatcaccac caccgacggt gcgaccgtgg gcctcccgtc gtcgctgaag gtgggggaga 8820

aggtgtccct cgccgccgag atctcctaca gcgacgggac gaaggcggtc aagcagacca 8880

atgccgtggg cctcacctgg acgtcctcgg acaaggccaa ggccaccatc gatggcggcg 8940

tggtcaccgg agtctcggca ggcaaggccg acatcaccgc ctcgatcgac ggcaagactt 9000

ccgaagcgct gtcgctgacc atcaacaccg ccgcctgacc aaccctcaaa ccctccgccc 9060

cggtcgtcct ctcgcgccgg ggcggagcct tgccacaccc gcgagaggtc aacttttctg 9120

cgagaggaaa ccatcatggc cgaggccaag aagatcagcg ccgccgagaa ggcgcgccgc 9180

gagacccagt ccgcgaagga caccggcacg atcaccgaca ccaccgtgca gatcggcgat 9240

atcgagttga ccgtgcccgc cgccgtcttc gaagacgact gggaattcca ggaggcgatc 9300

ctgatggcca acgatcccga tgccaccgac gaggatcggg ccagggcaag catgacgctg 9360

ttccgtcgtc tggtcggaaa ccgccaccgc gaagtgcttg accagctgcg cgacgagtcg 9420

gggcgtgtgc cggtgtctaa ggtcaccgag accgtcaaga aggtcatgga cgcggtcaac 9480

ccaaactgat gagcctcttc cagctcctcg ccacacattg ggaggagctg gagggggact 9540

tccaagaggc ctaccgcgtc gacctgcggg acttgtggcg tggtcggctg agcccggcgc 9600

gctgctgggt gctgctgaca caactgccac ccgggtctcg gctctggcgg atgctcggcg 9660

gccccatggc gtggggcatg gtcgagcgcg ccgtccgtga agagggctgg cgactcgcct 9720

cccagaacgc tggtaaggaa ctgcctcggc cggagccgcc tgcgccggga tggcgcgaca 9780

agcaggacga cctgcgacgc cgcgaagagc gccgtcttgc ccgcttcatg caacgccacg 9840

cagaacgcaa caactgaaca gtgcaccgtc ccgggaggtt tccatggctc tagatctcgg 9900

taccgcctgg gtgcaggtgt ctccgtcctt caggggcttc gcctccacgg tgaacaaaga 9960

ggtcggttcg gcagtgggcg gggccttcaa gtctgcggcc aaggtcggca ccaccgcgat 10020

cgccacgatc ggtgcggccg tcggtgggct ggcgctcaag ggcggcatcg accgcgccct 10080

gtcgatcgag caggcgcagg ccaagctgaa gggcctgggc cacgacgcag ggtcgatcac 10140

cgagatcatg aacgacgccc tcgcctcggt gaagggcacc gccttcggtc tgggcgatgc 10200

cgcgacggtt gccgcgtcga tgtcggctgc cggcgtcaag tcgggcgagc agatgaccgg 10260

tgtgctgaag acggttgccg acaccgccca gatttcgggg cgctcgctca ccgatatcgg 10320

tgcgatcttc gggtcggtgg cggcccgcgg caagctgcag ggcgacgaca tgctgcagct 10380

catgagctcc ggcgtgccgg tgctccaatt cctttccgac cagctcggcg tcaccaccgc 10440

cgacgtgtcg gacatggtgt ccaaggggca gatcgacttc gccactttct ccgccgccat 10500

gcagaagggt cttggtggtg cggcactggc tggcggcgaa accttcaccg gtgccatggc 10560

caacgtccgc gccgccctgt cccggctggg tgaggctgcc gccaagcctg ccctggacgg 10620

gctgcgcaat gtcttcaacg cgctgatccc ggcgattgat gccgccacaa atgcgctcaa 10680

gcccatcgcc agcgccctgg cgaaccgaat ttcgcaagca gcagaggcgg cttccgcctc 10740

gatcgggcgc ctcaccggct ccctcacgag catcacgaat ctcaatacag ggatgctcgg 10800

cgcggccttc tcatcgatgc tgccgatcat cggagcactg tcggggcagc ttggctcctt 10860

gcttggcggg atcccggtcg tcgggcaggc cttcgcaggg atcactgggc cggtgggatt 10920

ggctgccggc gtgctggtcg agatcgtggc ggcttcatcg tcgctgcgtc aggccctggg 10980

cacgctggtc ggggtcgtcg ggtctcagtt gtccggtgtg atgacgggca tcgtcgcggt 11040

gtttgccggc ttcaggtccg tgcttggtgc cgtcggtgac gttctggccc cgttcgtgga 11100

ccgtgcggcg gacgccgcca atgtggtcct gcccttgctg gggggtgcgc tgtcggctgc 11160

cggtggcatc ctgcagtctt ttgcgggttt catcgagcgc aaccatgtgg cgctctccat 11220

tcttgcgggt gcggtggttg cggccgcgac gagttggaag atctataccg gcgcgcaaga 11280

tcttgcgcgg ctggcaacga cgaagctcgg gctcgcgaca acggtcctga agggcaagct 11340

gtcatcgatg ggggcggcgt tcaagacgaa tccgttcggt gtcatcctca tggcgatctc 11400

ggcgctggtg ggggcgttct cgattgccta ccagtcctct gagacgttcc gcaacggtgt 11460

gcaggggatt ctcggctcgc tggcgccggt gttttcctcc ctgatgggga cgctgtcggg 11520

gctattccag caggtcgcgg gcgctgtcgg gccggtgctg tcgtcgatcg tctcgacgct 11580

ggcgtcggtg ttctcggcga tcggtcccgt cctgtcgcag ctggccggca ccatcggatc 11640

tgtcttctcg gcgatcggtc ccgtcctggc gtcggtcttc gggtcgatcg ggtcggttct 11700

ggcgagtgtc ttctccgggg tgatgagtgt cgtggcgccg atgctcaccg cgttgcagcc 11760

gctgttcacg cagctgtcgg cttcggcggg gcagatcggt gcggcgttcg gtcctgttgg 11820

tcaggcgctg tcgtcgtcct tccagcaggt cggtgccgcg ctggcgccgc tgctgccgat 11880

gcttggtcag cagttcgggg cgatcctgtc tcagctggct gcggccctgg ctccggtcat 11940

gggtcagttg ctggctgcgg ctgctcaggt gttgccgacg ttggcgcagg ccttcgggca 12000

ggtcgccggg gtgctgatcg ggtcgctggg tcaggctctg acccagatcg ctccgctgat 12060

aggccagctg gtgggggtgc tgatcgggtc gctgggtcag gctctgacgc agattgcccc 12120

gctggtgggc accctggtcg gggtggtcgc gcagctgttc gcccagctgg cccctttggt 12180

gggtcagctg ctggtgcagc ttgttccggt tgtcgcggga atccttgtgg cgatcgtgcc 12240

gatcgtcggg atgctgatta gtcagctcgt tccggtgatc gtcacgctgc tccaggtgat 12300

caccccgatt atcaccatgc taatcagcgc gctggtgccg gtgatccagg tcgtgaccca 12360

gctggtgctg gcgatcatcc aggcggtgat cccgttgatc tcggcgatcc tgccggcgat 12420

ctcggcactc atctcggcgc tgctgccggt gatcgtcatg atcatccagg tggtggcgca 12480

ggtgctgcag tggctggcgc cgctgatctc caccctgatc acggcactga ttccggtgat 12540

caccacgatc atccaggtgg tcatcacggt cgtgtcgaca atttggtcgg tggtcggggc 12600

ggtcattggc tggttccagt ccacggttgt gcccatcatc ggcaccgttg ttggtgcgat 12660

cgcgaacgct ttcggttggg tgcgcgaccg tatttccgat gcctggaact ggattaagga 12720

ccgcattgtc gccccggttg tcgagtggtt ccagtccacg gtggtgccga agttcgaggc 12780

ggtgcgcgac tccgtggtgc gggccttcga gacgctgaag gatggcgttg gtcgcgcctg 12840

ggatgcgttg aaggatctcg caaagaagcc ggtcgaattc gtcgtgaaca cggtggctgc 12900

cgggttggtg cgggcctaca actgggtggc gacgaagttc ggtgccgacg aggtcaagga 12960

gcctcatgtc gagttcgcca acggcgggtt cgcgggacgt gaggccggct tcgcgtcgtc 13020

gccgatcctg tgggccgagg ccggcccgga agcctatatc ccgttggatc cggccaagcg 13080

gacacgctcg ctggggatct gggccaagac cgggcagatg ctcggcgctc tacccatggc 13140

tgacggcggg atcatcggga acatcattgg cgggatcggc aacgccgctg cggcgatcgg 13200

caatttcatc aagtcaccga tcgagtggct catgggccgg gtccgggacc tgatcgatga 13260

tgtgggcagc tcaccgttcg cccagatcgc cgcgaagatc cccggcaaga tcgccgacga 13320

tatcggcgcc tgggtcaagg aacacatggc ctccatattc ggcggcggcg gttccggatc 13380

ggaagcgttc gacggctggt ggaacgcggc tgtcgccatc aatcctgata tggccccctt 13440

caagcagatc gccgccacgg tcgcccagaa cgaatccgga ttcaacccga acgtcatgaa 13500

caactgggat tcgaacgctg cggcgggcac gccgtcgggt gggctgatgc agttcatcca 13560

gcccaccttc gaggcctaca agtggcccgg attcgacaat tggatgggtg cggtcgatca 13620

gatcctcgcc tggtggaagt acgtgaatgc ccgctatggc gggccgttca atattcccgg 13680

aattgcctcg ctggcgggtg gcggcggata tgtcggctac gccggaggca ccctgaacgc 13740

ggctgccggc acggcatggg tgggggagaa cggccccgag ctggtcgatt tcggtggcgg 13800

cgagtcggtc tacaaccgct cccagattga cggtctggag gatcggatcg ctgaccggac 13860

gatttcccgg ctgcagcagc tgagggtggc gctgatcgtg gacggacatc agatgggtca 13920

ggtcatcgac ggccgcatct ccatggctgg cgctgctgca cacggatcga ggtggtgaca 13980

tggcgatcat tgcgacgcgc cgcgactggc ctgaggctcc gcaacgcttc cagtccgccg 14040

atgggcggct ggtggcggag ctggaccctg accggtgcgg agtgcgactg cgcggcaccg 14100

acctggaggc gtggagcgtc accctcaccc gtgatggcga ggtgatccac accggcgacc 14160

ccatggtcac accgggagga acaggaatcg cctacgacct gtctgcaccg ttggatgctg 14220

atgtcgtcta cgaggcgcac gcgggtgggg cggtgctcac gcaggtggcc gtccacaccg 14280

gcggcttgcc tttcgagtgg gggatggtga ccccgctggc cgaccccgac aagggcctga 14340

tgctacggac cgtcgccgac acccccacgc tgggcaggtc ggcacgccag aagctgtctg 14400

cggtgccctc atcgaggctg caggcaggtg gctgggacgt ccccaccgac gcggcacagg 14460

gatggacgtg gctcgcggga ttccccgacg cctccaaagc gctcgccgag cgcgacgcga 14520

tcatggaggc cctatcgctg gggccggtct acttccggcc cgaaacctcg atcggcttcc 14580

cgcccatgtg ggcactgccc ggcgacgtgt cagcgaccaa gcagggcgac gcctggacgg 14640

tgtcgtgcac gctgacgccg atcaccgctc ccgcgaccgc cgacctgccc gcctgggcgc 14700

ccggcaacag ctatgcgcgt gtggcggcca cccgggggag cctcgccgag ctcgcccgca 14760

catccaagac attcctcgag ctagtggggt tctgatgatt gaagtatcca agcgatgggc 14820

ctcctcagta ggggccggtg cacgctggtc ggtgatggtc tcctggtcct ccgacggagg 14880

ccagacctgg catgacgtgg tgcccaccgc ctgctcggtg gacgagtcta ccggccagca 14940

ggtgcggtgg aagctgtcct gcaccctgcg caaggccgac gccgagggcc tgaccgtctt 15000

cggttgcagg gcgcgcgtct tcgtgtcgat gcatcacacc gacagctggg aggagacgat 15060

ccagctcggc gaattccgca ttgacaccac ctctgacacc accctcgccg ggccgtccgg 15120

tgcgcaggtc gcggcagttc aggtgagcgg ttcgagctgg gagcagcagc tgatggactc 15180

gcggctggtt gagccgcgtg aggtgtcggg tgccgcgatc gatgtgctcg gcggcctgat 15240

ccgggaggtg ctccctgacg cagagatcgt cttcgacggc gggatcgatc cgggccgcaa 15300

cattccggcg acggtggtgg agcgtgaccg gtgggccttc attgacggct cgaattcgtc 15360

ggagacgtcg gtggcgcgga tgctcggcgc ccaggtctcg accgacgcac ggggcgtgtg 15420

gcatgtggcc ccgcctccgg tgctggacgg gacggcggcg tggacgatcg aggccggcaa 15480

gggcggtgcg ctcctgtcgg cggtggccag cgaggaccgc tccacgatcc gtaacgccgt 15540

catcgcgcgc ggcgagtcaa ccgataagag cgtgccggtg ttgggtccgg tgaccgtggc 15600

tgatcacaat gcgtggtcac caaccaacgt ggacactccg gtctccaggg gcggcttcgg 15660

cacagtcccg atcttctaca cttcgagcct tttcaccgac acgacgcagg tggaggcggc 15720

agcgaaggcg atgctgcagc cgcgcctggg cgtcaaacgc accctggacc tgacaacgct 15780

cttcgaccct gccaaacgcg ccggggatgt gggtgtggtg cagaccactg atggtccggt 15840

caccgtcgtg ctcgaatcag tgtcgtgcga cctggtggcg gcgtcgatga cctgccagac 15900

gcgcggcacg accggcaccg agctgatcac gaccgaaacc acgacaacca ctggggagaa 15960

gatctcatga gtgcaccaga cattgccctg caaggactga tcggggaaga caccgagcag 16020

gtggcgcttg cccaggtgct cggcgtgggc gtcgacgggc ggtcggtgcg tgtccagcgc 16080

ggcactctca cccacgaggt ccgccggctc gatagctaca agccttcagc gggagaccgg 16140

gcgctgctgt tacggctatc tggcggcgaa tgggtgctga tcggcgccct cgcctgacct 16200

tgacgaccta acctctgaca acctgaaaag gagccctcca tggcaaccgt ctatggccct 16260

gacaaattca ccgtcccgac tggtccggac gcaccggacg tgccggcgac gatcatcacg 16320

ctgctggact cgatgcgtcc ctcgctgatc gggcatgcgt cttcgatcgc tgaccgcacc 16380

gcgaaatatg ggcgggcatc cgcgtcgagc attcaggcgc cgaagggcac agtggtggtg 16440

tctgccgagc tgaacgcaat ttgggtgaaa acatcggaca cgctggatga gtgggcgacg 16500

atcattcagc actcggatga ggtggcgacc gtgtcggtgg tgtccaccca gtccgaccag 16560

gtgaccacgg tccagaagtt cacgattccc gagtcgggca tctatgcgct gtatgcatcg 16620

atgaatgacc agaacggctt ggatgtcgat gggtcgatcc gtgagataca tgttctggtg 16680

aacgggacct ggaagttcgg tgggatcttc ccggcgagca agttctggct ctggtcgggt 16740

tcgcggacga cctttctcaa taagggcgac acctatcaga tcgactttat gcaacgctca 16800

ggcggggaga ggtccctgaa ggtaacgctg tcttatcaaa ggattttgta atggcgacgt 16860

gggattacgg gtatgcgccg gctgatgtgg tgaccgatgc ggccggggat gtgctggccg 16920

gcatcgaact gcgggtgtgg gacgccgagg tggcagggaa agccgtcgcc gtccagcagg 16980

accgtggcga cggatggaaa cccgcgtcaa gagtcctcac cgacgacgtg ggccgctacc 17040

gatttcgtgc cgaagcgggc cccacggtgt gggtggagga cgtgtcaggg cggcgctggc 17100

ggatggatgc ctggcagacg ctcggcacga tgatcgactc cgcacagagc gccaccgccg 17160

cggccgagtc ggccaactca atcgcccacg aagccatgtc agtcgcccaa caagcccaga 17220

cgtcggcgaa ggccgccgcc gactccgccg ccgccgtgca gggggttgcc ccgtccgacg 17280

cgaatgtgtc gccgatcatc accggcgggg cgaagactgc tgaggcggtg cggaaggcgg 17340

cgctggctgc tttcccgacg accgggccga cgatcttcac gcacttcttg acgcgcgacg 17400

aggccctgca tgtggcgatc tccaccgacg gtgtgacggt ggaggacacc ggcctgcggt 17460

ggaagccgaa gaacgacacc accctggggg agtgcttcgt gcgcgaccca tcggtgtgtt 17520

tctggaaggg tgcctattgg gtcgccttca cccggcccac gaagggcggg ggtgacgctt 17580

tcgggacgac caagtcgttc ggactgatga agaccacgga ctggcggacc ttccaggagc 17640

tcccgccggt cgtgatgccg agtcaatttc agcagacgtg ggcgccgcag tggttcatcg 17700

gctccgacgg ggtgccgcat atctttgtgg ccctcggcac caccaccacg cccaacgcgt 17760

acttcaccca gtatgagctg cggccgctcg atgacgcgat gacgtcctgg tcggacccgg 17820

tggtcatgtc tggactgcca gcgaattgca tcgatgtcgc ggtgatcgag gacgccggta 17880

ccttccacgc ctttccgtcc aaccagaaga cgtcaacggt cgagcagtgg acgtcaaccg 17940

ggctcaccgg cccctacacg aagctggcgg ccagcgactt tcccggtgcc ggtgtcgaag 18000

gaccccagcc agtgccgctg aagacgggcg gctggcggat ctacgtcgac aattacgcgg 18060

agaccgactc gatctatttc gccgagagca cggacttgct gcattggtcg gcgctcaggc 18120

cggtcaccct gccgatgcgt cacgtcggcg cggtcgcggt ggactccttc ggtgcgctac 18180

gcacccgcga gctgtggcag ccgaacatcc cgggcatgag ggggatgggg gcacccttct 18240

ggggcgtacc cttcgccgcg gggaacgtgc tgaaggaatt cgcgcagatc gtgtccatgc 18300

gcaccgacgg cggcggcgaa atcgatctgg caaaggcggc cacgctgggc ttcaccggca 18360

tcgattacat ctcggcgacg gctgtcgcga acgtcgagat tctgcagatc gagcccgaca 18420

ttcgcgctgt cgacagcatg atccacggcg tcgccctgcg aggaccgagt acgccgcaga 18480

tcgatacaga cgtgaaggtc gcctggcggg tgctcggctg gggcgatccg agcacgccat 18540

gagcagggac gctgacgtga ccaagcaggg atccttgcct cggcgggtct gggacatgct 18600

ggcagagccg aagtcggtga cggtcctcat gacgattgcc tacgcggcgc tcgtcgcgct 18660

cggcttctgg gcgatcgacg acgcctccac gatgggggtc cgcgacatga tgggcggcct 18720

gctcatcgct ggtggcgtgt gcgggctgat cggatgcccg tggggccagt ggtggatcga 18780

gcgcgccggt ctggtggcga tcggtgccgc tttcgcggta cacctgtctt tcgtcgtggc 18840

gatctccccg cccgacggac cgtgggaagt ggcctcggcg ctggggctgc tgcttctcgt 18900

ggcgacacgc tggatcagga tcaggacgct gccagccgac ccgacgctgc ctcggcccgg 18960

gcctccagag gcgggggatg aatgaatgac ttccagacct ggatcacagt gctgggcggc 19020

gccggattcc ttggcgcgct cgtcacgctc atcaaggggc tggttgggtg gcgcaccggc 19080

aagtccggcc gcaaaatgag ggccgcccac gacgccatcg actcgctgaa tctggcgggc 19140

ttgtgggctg aagcctactg gcacgctcgc ggctattgcc gcagccacca tgaatggacc 19200

agcgattacg ccgacggcta tccaccccca cccgacgaca ccaacacccc tgactgagcc 19260

ccgccttgtg cggggctttc tcattcctca aagacttgga gacattcatg gactggacca 19320

atctgaacgc tgacgtgacg aagctgatgg gcgtgcactt cacccccgga cgtgaaggca 19380

ggacgatcga caagatcgtg atccaccaca acggcggcaa cctgagcatc gaccagatct 19440

ggaatgtgtg gcagacccgt gaagcctccg cgcattatca ggtggaggcg ggtggccgta 19500

tcggccagct cgtcaacgat ttggacaccg cgtggcatgc cggcgactgg gacgccaacc 19560

tgacctcgat cggcatcgag catgccgacg actcgaccga cccgtggcat gtgtctgatg 19620

ctgccgtcga tgccggcgcg cacctggtgg ctgcactgtg tcgcggctac aaccttggcc 19680

ggccggagtg gatgcgcaac gtcttcccgc actctcagtt cacgtccacg tcgtgcccgg 19740

cgtcgctggc ccgggaccag ctcggcgact acatggggcg cgcacaagcc tacttcgatg 19800

gcgcgccggt ggctgcggtc catcagtcgg tccctgcccc cgccccagcg cccagccgtc 19860

atgtggacct gcccgcgtgg aatctccccg agggcaactt ctacggcctc gtcagcggcg 19920

gaaacgactc ccacggcggc ttctatcccg ccgagcgtcc cgctgtgagg gccatccagc 19980

tgtggctcat ccgtcacggc tacgccggcg cggtgcctga cagttgggcg gacggcatct 20040

acgagcagcc gaccgccgac gccgtgaccg ctttccagca cgccgagcgc cccaacagca 20100

cggaccggtg gggcgaggtc tgggccgacg acctggccac catggccgcc aacaactgac 20160

aaggagctga tgccaagtga tctggactct cgcattctgg aagggcgcag gcgagcgcgc 20220

catcaaaacc gccgcgcaga ccgccgtcgg cctcatgggt acctcgacgc tcatcgaaca 20280

ggtgccgtgg actgtcgtcg cctccggcac cgccatggct gtggtgctgt cgctgatcac 20340

ctcgatcggc aacgccgact tcaccgccgg cgtccccact accgccaagg ggctcgaggc 20400

gacgaccgtg ggcaagacgg acaccacgcc cgtcacgcca ccggcgcgcg tcgccgaaga 20460

ggtcccagcc ggcttcgtcc cggacacggc cccggatccc gtgccgaccg tctgacctga 20520

gggggtgacg gcgaccctgc gccggatagc cactcaagca acctgagcga cacaagaccg 20580

cccactctga ccttcgcggg tcggagtggg cggccttttt gcgtctcagg ggcgcagatg 20640

atgactcgtc gtctttaatt ctagcagtac gcgttcagcg tcgccagacc atgactttct 20700

cggctgcctg gagcggcgca ccttcggggc ctttgaggta gggggcgatg tagatgagct 20760

tgcgcagtcc atgcttgggg ccgtgcgcct ggtgggtcca gtgcccgcgg accatgaacc 20820

gcacggtgag cttgtgcccg gttccgtcgt cgcggtcggt gaccacggtg cgcacgggac 20880

gcagatcgac cagggtgacg tgacggtcgg ggcgtggcgt gcggggcctg tgctcggtgc 20940

ccggggcctt gccggtgcgg gagtcgatcg tgcgccgctc ggcgacggtg ggggtgtcca 21000

tgagcacgct catcgccatc agcagcgacg cggacatgcg ggcctcgggg ctcagcgtgg 21060

catccagatc ggattcgggg cggatgagaa tcgacaggat ctcgacaagg ggaccgtcca 21120

cgtcagcgaa gccgggcgga tagtcgccca gccgccccag cagctggatc atggtgcccc 21180

caccgggagc gggaagccag gcgatggccc acacgggagg attcccctgc caggtgcggc 21240

cgccgggcag gtcgaaggtg cgcggcttgg gccccggcag gggtttggcg aagcaggcca 21300

gtccggtcgg ggtgatcagc tggctcgggc tccactcggg cacgtccagg gcagcatcca 21360

gagcgagcgc cgccatctcg ccactcaccc agaacagcga cgcattaccg agccgctcgg 21420

cactccaccc gaagccggac atcggcagtg ccttgtcgcc catggcttca gccaccgcat 21480

cagggtgggt ggcggccagt tgctccaggc gctcgtcgag gtggcgggag tcccgcacaa 21540

agcggcgacg cagcccgggc actccgcggg gtgtccacgt ccagctctcc ggggctgcca 21600

tgtcaggcga cgactcgaat gcccaggctc cgccacgcct cggcggtgtg gtcgagcccc 21660

agatcccacc actgggtcag cgcgtgttcc atggcgatcg tggcgcaggc cgacagctgc 21720

gaggccttgt cgcgagccga cagcaggtcg gcgctgtcgg tgccgtcggg cgtgaagtcc 21780

tgcagggcga tagtgatgtc tccgccggga agctgggaga ccgtgtcggc ggtcatgcct 21840

gcctgggtgg cgtcggcgat gatgcccagc tcgaccagcg tccagccggt gcgggccagg 21900

tcgatgcgct gggcctcgcg ccacaggtcc atctcggtgc gggtgcgtgc cgacaggctc 21960

gggggtgcgg ccacatcggc tccgcgtcgg gccatccatg cggccatgtc gtcggtgggc 22020

cgccaggtga ttgtgctggc catgaagatc ctcctcggag tggaaagtgg aaaggggagg 22080

ggccggagcc cctcccctga tggttgatgt ggtcagcgga tccaggtgaa gggctggtcg 22140

ccgatgatcg agcggaccgc gaggtcgtag ggggcctcgt cctcgctgga ctcgacatcg 22200

accgcgtcga gctcgacacc gtcgcgggtg atggtgatgg tgtcggtcgt ggtggcccga 22260

ccgtcgatca ccgtcggagt gttgatgctg gtgaactctg cggtgatgcc gaccagctcg 22320

tggtcggcga ccgcttccca gaaggcgtcc tcgtcggctt cgatggagaa gtgcacgctg 22380

gaggcgatcg tcgcaccctc ggaggtctcc tcgctgtgga gcgtgaccag ctcgtcggcg 22440

atggcgtcga ggtcatactc ggcgcgggcg tcggcgactg cacctccggc ctcgatggca 22500

tcgatgatag aggcgatggc ctcgccgcgg gtggagaagg tagtgtcggt agaatcggtc 22560

atgatcctgt ccctttcagg gtcttggcct catcggggtg cttcccggtg gggcctcttg 22620

ctttgtcctt gtgacaccca ctgtcgtcac agtgttggaa catgtcaagc cagtgggcca 22680

cctttcttta aagagatttc agcgggcgac cgcgccgcct cggacacctg ggcccagtag 22740

ccctggaggg cgcactgaat caattggttg tcaattggtt gtcaaacctg acccgtcgac 22800

ggggagtgag gaggtggtac cggctgatct acgcctgaaa cagatggagc gggcgacggg 22860

aatcgaaccc gcgtgtctag cttgggaaac gggcatcgtg ctagtctggg gaccgccgaa 22920

atgacgattt caggcgtaaa ccggcctccg gtgtcttacc ctgatagctg ggtgatagca 22980

ccgaattggt tgtcagattg gttgtcagat cgccccagga ggatggtcgc attgtcacgc 23040

gcaagctacg gggacggcac ccagccgacc cggcgttccg acgggcgctg ggcagcatcg 23100

gcctatgacg gctggcaggc gaacgggaac cgccggcgcc gatgggtgta cggccgcacc 23160

caggccgaat gcaagcggaa gctgcgcgac ctgaagcggg agatctggtc agacacccag 23220

cagatgaatg tgaaccccag ggagaccgtc aagagctgga cggcatcatg gctggacgac 23280

taccgatcga ttgccagacc aacaaccttc gccaccgacg agtccatggt gcgcaactgg 23340

atcgtcccag ccatcggtgc ccggcgcctg tccgaactga cagcgcgcga cgcctcgaag 23400

ctgcaacggg tctgccgaga cgggggactg tcggcgacaa cgtctcacta tgccgggctg 23460

ctcctgcggc gcatcctgaa ggctgcccgc gcgaacggct accgcatccc cgactccgtc 23520

atgctggccc ggatcccggg catcggcgca tccaacaggt ccgccctgag cgccatccag 23580

gcggccaacc tgctctcgac ggcaaacgca cgcgacacct ggccggagcc gcccagcctt 23640

cccgacctgc cctacggggc catctcgaag ctcgcaccag cagaagcgca gaagcgtgaa 23700

caactcaaga tggagcggtt ggaatggact gccgcccaaa acacggaccc ctccaggtgg 23760

gctgccgcac tcatgcaggg acttcggtca ggagaggctc gaggcctcac gtgggatcgt 23820

gtcgatctcg ataaggggac gatcaccatt gatcgtcaac tccagcgcat caagcccgac 23880

gcggcgcttc caccgggata caaggtcacc cggctggaag gcagccactg cctcgtggca 23940

ccgaaatctc gatcagggat ccgccgcgtc ccgatcgtcc cctggatggg ccaggctctc 24000

acccgctggc gcgacataca gggcgacagc cccttcgggc tcgtgtggcc gctgcccacc 24060

ggggcgccgc ccacgcgggt ccatgacctg cgggcatggc gtggactcca gcgcgtcgcc 24120

ggggtccaca aggaggatgg aaacctctac gtcctccacg aagcacgaca ctccaccgtg 24180

tcgctgctgc ttgctgccgg ggtcccggaa tcagtggtca tcgcgatcgt cgggcatgca 24240

agcttcgcgg cgaccgagca ctacgcccac accgacctcg aagcagcacg cgccgccctc 24300

atgaaggtgc aggaccgcct cgggctggag ctcgagagct gagcatgcaa agagccgccc 24360

accggaccaa tcgcggtctg gtgggcggct cttttgcgcc ttagagcacg tccgtcacca 24420

cgcctggaag ttgctgacga cgggtgcctg gtcggtgccg gtcacgtcgc agtgaaccgt 24480

gtatttggcg gagccgacgt cggcgccgat gttgacgttc cacagatcgt cggtcttatt 24540

gagggcggcg accgaatcga cggttgagtg gaccttgatc ttaagcgatg ggtattgctt 24600

gcccaaggca tccctcgcat aggtgccaca gccagaggtt gcgccggtca tggtgagtcc 24660

agtggtcgtt gcctcgacgg gtgtgggcgt ggcggacgca gtgggcgtct tcgtggccgt 24720

cggcgtcttc gccgctttcg gagtcttgga ggctgatgat gacgatcctg atgtctgggg 24780

atcgcacgcg gtcagcgcgc cggcgaggca gagtgacgca agcagggcga tggggacgag 24840

cgccttgcgg cgcatggtgt gggtcattcg ggttccttgg ttggttggtt cacatgctgt 24900

ccacagcaac ctagccgcgg aacctgcccg cctgggggtg atgagggcaa gtggcaagaa 24960

ttacatcgat gggattctcg ccaccccctg aagagcgtgg gtggcgaggt ctacattcgt 25020

acgcatgtac gaaacatgga agcctctcgg acacggctcg atctctggtg gatcggcccg 25080

cacaatggag tgcactgaag tagcggagtg ggcagagcgt cgtgcgcgtg ggtggggctc 25140

agcgctggta tcgcgcctgc gcggcgtcca tgaagacgcc gggttcaaag tcgagcacgc 25200

gagcaagctc aaagaggagt gcgacaggga gatcccgctt gccctgctcg atcctgatga 25260

tggtggattc gctgactcca gcgagtcgag cggtctcgac ctgggttaag cccttggcgg 25320

ctcgctcggc tcgaagctgg gcggcgatcg cggcgcgaat tgcatcacgc ttgctggcct 25380

ggttctggtc catgctgtca gcatagccgc cacattggac agttttccgg tccgattggg 25440

atgctcggca cttgcatctg gccatatggc atggcaagct gtccatatgg ccagttcaga 25500

catcaacctg gaggctgcgg acatgatctc cgccgccatc gagcgaagcg acaccagtcg 25560

ggctgaagtc gccacgctga cgggaatccc gttgaccact ctgcgtcgga agctcatggg 25620

ccgatcgccc gtcaacatcg aggacatctt cctgatcgcc ggcgcgctcg ggataccgcc 25680

tgtgagtatc acgcccgacg ttctcacgag tgaagccgcc gcctagcccc caaacagaag 25740

aagcccccgc ctgctgtcac agacgggagc caaccaaagg agtttccaat gagcattcta 25800

cccttcgact accacggtca ggaagtccgg ttcatcaccg atgagtccgg cgagcctcag 25860

gtcgtcgcgt cagatctcgc gaaggccctc aactatcgga acgcacccga catgatgcgt 25920

tccatcgacc tagaggaaag gggtacgcgt ccggtgcgta cccctggcgg tgagcaggag 25980

atgctcacgc tgaccgaggc cggcatgtac caagccatcc tgcaacgcca gacaggccgg 26040

atggtcgacg tcgcccaacg agccgctgtg aagcgattcc agcattgggt tacccacgag 26100

gtgattccct cgatccgcaa gcgcggcatg tatgccactc cggatgcagt cgaggcgatg 26160

ctggccgatc cggacgttat gatccggacg ctcaccgagc tgaaggccca gcgggccagg 26220

gtggcccagc tgcagcccaa ggccgactac gttgacgcct tcgtggccga cgaggatctg 26280

cggctcctgc gcaatgtggc caagtcgatc ggagtgcagg agggcgccat tcgcgacgcc 26340

ttgctcgcac acgagtggat ctacgcggag gagtcctcgc gctggtcgaa ctctcagggc 26400

tgcaaggtca tcgagcaccg ctattcaccg cgctctgaca aggcccgata ctttcgcccg 26460

gtcccgaatc accaggcacc ccgatttaag ggcgaggtaa tgcacaccct gaaggtcact 26520

ccggcagggg ctgaggcgat ctccaagatg gcaaagcgct ggggcctcgt cgtccaggag 26580

gtggcggcat gacctcgact ctcaccggca acatcatcgc cctgctgatc gtggccggcg 26640

tgatcgtcct cgcgatgggg gtgcgccgtg aaggtcgatg acttcgacga tgtgcgcccc 26700

ctgacgcaga aggacgtcgc cgagctactc cacgcaagcg tcggttacgt gcgctcctgc 26760

cgcctggcga cgaagccgaa aggccgggtc ttcccgatgc ccggctggaa gaccgacgga 26820

aagcgctatc tgcttcccgc ttggcggttc cgcgagtggg tcgaaagctt gcccgatgcc 26880

tagcccgcgc cgcttcctaa tcctgatcgc cctgggtgcc gccgccgtcg gtttcgcgcc 26940

ctcctcaatt caatttctct tcatggccgc gcttgtgctc ggcctcacca tcacatgcct 27000

caaggagtcc aaccatgcct gacacacagc cccgtcgtgc gcgtcgtcgc acgctgtccg 27060

agatcctcgc ccccgcgccg gcgccccgca gagcggaggc aacggcatga ggccaccagc 27120

cgttgaaacc cctgatgtga aggcgccggc cacgcctgct ggttcccggc tcttcaaggc 27180

tgtccgtcct gacggcttcg acttccacag cgggactgtc cggtggctcc ctgctgatgg 27240

cgcaccgatc ccggagggcg ggtggcttgt cgagcatccg catcctggtg aggttggcag 27300

ctgggatgca gctttttatc tgtcggcgtc gtcggtggag acggactgca caggtttcca 27360

gtggcctgct ctcctcctgt ccgtggagcc cgtaggtgcc atgtggaccc ctcgccccga 27420

caaatttcct cgcaagcggg ccgcgcacgc gtggcgcgtc atagaagagc tccccgcatg 27480

gcggcttttc ggtccccagg ggcggacggt cctggacatc atcgagcaaa ccgctcatct 27540

gaccaaacgc cagatcgcgg ccctgaacag ggctctggac gccgcacggg acaccgtttg 27600

ggacgttgct tggaacgccg cgtggcacgc cgctcgggtc gctgctcggg tcgctgctcg 27660

gggcgctgct cggggcgctg ctcggtacgc cgcttgggac gctgctcggg gcgctgcttg 27720

gtacgccact tgggtcgctg ctcggggcgc tgctctcgga tggctcgtca aggacctgat 27780

ctccgtcgag gacttccgca ccctgacggg cccgtgggag caggtcatgg gtccgatcga 27840

ggtggcggca tgaaccgcac ctatttcaag gccgttaggg cggacggcac tgacttctac 27900

accggcaagg tccgctggct gcccgatgat ggcgcaccga tccctgccgg gggttgggtc 27960

gttgagcatc cgacgagcga acgcgtgggg gacgacgccc gcacctatct ctcggtttcg 28020

acggtggaaa ccgactgcgc cgggatgggc tggccgtgcc gtctcctgcg ggtcgtcccc 28080

gacggcagac aggtgagcat ccctgaaccc gtggggctgc ccagcacgag ggcctcgatc 28140

aggtggcgcg tcatcgaaga gctccccgca tggcaggcgc ttggacccca ggggcgcgag 28200

attgaggcgc tgctcggaca ggttgagagt ctcacggagg accagaccct cgaaatgtct 28260

gccgctcggg gcttcgctcg gggcttcgct cgggacgtcg cgcggttcgc cgctctggtc 28320

gcctctcggg gcggtgctct gaacgctgcc cagggcggtg ctttgggcac tgctctgaac 28380

gctgttcggg acgctgttct cggatggctc gtcaaagatt ttatctctga tgaggaattc 28440

cgcaccctcg tgggcccgtg ggagcaggtc atgggtcggg tgatcgcatg atgccgatca 28500

ccaagccgtg cgcggttaag gacatgccgg agggcgagta tcactcggat ccctgcgtcg 28560

agccgtccct gtcgtccacg atggcgaaaa ccattgtttc gggtgaggct ggcccggccc 28620

gtctgcgaga gatcatgtct cacgggcagg aacataaggc cgtcttcgat ttcggcagcg 28680

ccgcgcacga gaaggtgctg ggacgcggcg ccggtgtcga ggtgctggat ttccctgcct 28740

ggaccacgaa ggcttcgcgt gaggcgcgtc aggccgtgtg ggatgccggc ggaactcccg 28800

tgctggcgaa ggattccgcc caggtggatg cgatggctga ggcgatcctg tccaatcctg 28860

tggcaggtga gctgttcacg cgcggggctg gttctcctga attgtcgatg ttcaccattg 28920

acgaggagac gggacgctgg cagcggggac ggctcgactt cctggcggac cgcaagacca 28980

tcgtcgactt caagacatct ggacagtccg tcgagctgcc cgactggatc aagcacagct 29040

ggcagttcgg ctaccacatc caagccgccg cctatatgga ccaggcgatc tcgctggatc 29100

tggtcgatga ggacgccatc ttcctgcatg tcgtgcagga gacgaagccg cccttcttgc 29160

tcgcgatcta tcaggtttca gctgaccagc tggccgaggg caggcgtcag atgcgtcgtg 29220

ccctggacct gtgggaccgc tgcctgaccc tcgacgaatg gcccgcgatc cctgcggtga 29280

tccaactatc caagctgccc gattgggtgc acaccactga tgacgaaaag gactcctgac 29340

atgaccgaaa ccacacctag caccgacatt gaaaccaccg cccccacccc gtcggggtcg 29400

atcgcggcgg tcggctccga gacggcaggc ctgacgcttc agcagaagct cgactatgcc 29460

tctgccctgg ccgactccga gctcctgccc gccgcctaca agggcaagcc cgcgaatgtg 29520

ttggtggcga tggagtacgg cggcgagctg ggcatcggca cgctcgtcgc ggtgaaccag 29580

atcacggtga tcaacggcgg cgtctccatg gaggcgaagc tcatgatgac gctcgcccgc 29640

cgagccgggc acatcgtgcg cctgtccggc gacgacaagc aggccacctg catcatcatc 29700

cgcgccgacg atcccgggca cgaatcggtc gtcacttggg acgaggccaa ggcgaagacc 29760

gccggactgt ggggcaaggg ccactggcag aagaacccgg gcttgatgtt gaagtaccgg 29820

gcggcctcgg agaacatccg gctcacctgc cctgaggtgc tggcggggat tgtctacaca 29880

cccgaagagc tcgatgagcg caccgagcgt gcaggccggt ccacgatgcg tgtccatcag 29940

gtcgtggccg agccggagaa gaccgctgcc tacttcatga aggccctcca cctgaacggc 30000

ggccagttca aggagtttgc ccagcgcgtg ctgggacatc cgttgaagag ctgggaatcg 30060

ctggccaagg cagacaagca gcgtgtcctg ggcgctctcg ccagctggga gaacagcggg 30120

gccgatccca ccactggcga ggtcctcgac gccgagccgg tcgagggcgg tgcggcatga 30180

gcaccttgcc tgcggatgct gccgagaggt ggcagcagtg ggatggcctg gcccgcacga 30240

tcctcgccct tcatctcggc ctgactgatc ttgagatggt cgagctggtg ggcgggctca 30300

tcggtgccgg ctggcatcag gatgggccgg tggagtcatg agctggcccg aggagcacca 30360

cgacgtgtgg gcgggtgtcg aggacgccat ccctgagtgg gtgagcgaca aggtggcctg 30420

ctcggtgcgg tcggatgccg attggaatgc cgacgaggac agccgcaagg ccgtggcggc 30480

ggtgaggatc tgcgagcggt gcgccttaac cgagcagtgc ctggattggg cgctggccca 30540

ccacgaggcc ggcatctggg gtgggctcac cgcctccgac cgcgagcgca tcgagcgtgg 30600

cgcgccggtg cggcgggtcc gcgagattcg tcggcgtcgc acggcggtta ggcaggtgca 30660

ggagtcatga gcgcaccact gaccaaggcc cagaaggtcg cggcggtcgt cgagcagctg 30720

ttgcgtggcg gcgccgacac cagcacgctc ctggaggcga cgggggccga ccggcccgga 30780

cgattgcggg acacccttcg ccgcgctggc cgtgacgacc tcgccgcccg gatcatcacc 30840

accgaccggg cagcccagcg cagacgggaa gtcatcgagg cggtcgagaa gctggtctgg 30900

gtggacaggg ccgacgagat cgccgccgaa ctcggctaca gctcgcgcta cggcctgcaa 30960

cagtccttgc gcggctgggg gcgtcgggac cttgccgatc agatcgtgct gacccgcgag 31020

acgcaccgcg acagggtcat cgctgacgtg gaatggatcg ccggtacacg gggccccgag 31080

gatgtcgccc gggcgaccgg ataccgcaac gcggcggcgc tgcaggccgc cctgaccggg 31140

tggggccgca aggacctcgc cgaccggatc gtcggagcat cacgcaacga cacgggccgc 31200

ttccgcttca catggagggc cgcatgagcg ccaaccgctc ccgccgcgcc acgtacaacc 31260

acacggggat cttcgtccat ctgcgcgaag ccgccgagcc gtccgaacag ccaccctccg 31320

accagacatg cccagccctg catgtcatcg ccggactgac accctgggcc gaccaccagc 31380

cccgccacgc cctcggcgtc gacgggcgat gccggcactg ccacaccacc atcaaaggaa 31440

acccatgatc ttcaaagaca ccacgatcgg gccgctcgaa acacggttca cctggtcgat 31500

gaggtgcgac cgctgcggga cgccgctcga ctggctcgtc gccgcttcgt gcaagaccga 31560

gcgtagtgag gtaatcgccg tcaagttcct gagggagcgt gcccgcgatg gcgggggcct 31620

cagagagtgg ggggagctgg acctttgccc ttcatgcttc tcggtgatgg acgcatgatt 31680

accaccacac aactcggaga agcagaccgg tggggccgtg gcctccaagt ccgctcgatc 31740

ctgtgcaacg gctgcggcat agctctggcg accgacatcg gccttcgtgg agacgccacc 31800

gccctccaag tgcaatccga cctgcacgcc cgagcacgca ccgccggctg gacacacccc 31860

gcctggcgcg tcgacctctg cccgcaatgc accaccacaa ccaaaggagc atgaccatga 31920

aggccaccca gtacgccaaa tcgaccgacc ctgaagtcat cgccaccatc gaagagaacg 31980

agctgtcacg acgggcatgg atcgacgaca ccaaggcgtg gttcggcaag acgatccgga 32040

caggaatccc gggcgccaaa ttgttcctct tttccacccg gaccgctatc aggctgttgg 32100

ggatcgtgac gtcggacgag aagaagcctg ccgggtggaa gttctgctgg cgttcacgct 32160

ctcggttcga gccacgaaag aacaatccct tgcgcgccac atgggacgca cgccggtggc 32220

aagcagcgtc gatcccaggt ctgcccgtgg ttctcacgtc ctccgtgtcg ggagagttac 32280

agagctggtt gaggatgtat ccctgcccct tcatctctag tggtgccgca tggctggacc 32340

tggagcacat gcctgaccct gacagtccgc acttcggacc gcagtggact gaagtccgtg 32400

catcgcaggc aatggcagcc aaggaagcat tgaaggacgc gtcatgagca ctccgggatc 32460

actgcgcgcc gcgctcgacc agctggacga gatcggcatc gccgaccatg tgcagtcctt 32520

ggaatgggat cgggccggcg cccgcaccac agcctggctc gagacctgcg gcgacttcgc 32580

tgcggcctgc cagtggggcg atgccgcggg cgaatgggtc acgtgggaca tcaccgacgt 32640

ggccgaggcg gacgtcagcc cccggctgcg cgtcaagcac atgcacctgc gagccaggcc 32700

ctgtgctgat gcgcccgcga aggcggtggc ggcatgagca aggcccttga cccactggat 32760

caccttc 32767

<210> 23

<211> 29768

<212> DNA

<213> PAC7 phage

<400> 23

tcgtacggct tagtgaaata cctccctttt gttgttttat cgttttgtcg actttttgtt 60

tggtggtgtg tgtggtgcag cctgagcttc ctgatagtcg tgattggtgt ggggagacgc 120

gtcggtggtg gtgtgtgtgg ggcgaggatc cgcgtgccgg gtttgtgtct gatgaggagt 180

ggttgtttct catggatgct gcggtgattc atgatgtggt gtggcgtgag ggtcgcgcgg 240

atttggtggc ttcgttgcgt gctcatgtga aggcttttat gggtatgttg gataggtatt 300

cggttgatgt ggcgtctggt ggccgtggtg ggggttctgc ggtagcgatg attgaccggt 360

ataggaagcg taggggggct tgagtaggtg tctggtgttg ttgggtctca ggttcctcgt 420

caccgggtgg ctgtggcgta ttcggtgtct gctggcgggg atgctgggga gcttggtagg 480

gcttatgggt tgacgcctga tccgtggcag cagcaggtgt tggatgattg gcttgctgtg 540

ggtggtaatg gcaggcttgc ttcgggtgtg tgtggggtgt ttgttccgcg gcagaatggc 600

aagaatgcta ttttggagat tgtggagttg tttaaggcga ctattcaggg tcgccgtatt 660

ttgcatacgg ctcacgagtt gaagtcggct cgtaaggcgt ttatgcggtt gcggtcgttt 720

tttgagaatg agcggcagtt tcctgacttg tatcgtatgg tgaagtcgat tcgtgcgacg 780

aatggccagg aggctattgt gttgcatcat ccggattgtg ccacgtttga gaagaagtgt 840

ggttgtccgg gttggggttc ggttgagttt gtggctcgta gccggggttc tgctcgcggg 900

tttacggttg atgatttggt gtgtgatgag gctcaggagt tgtcggatga gcagttggag 960

gctttgcttc ctaccgtgag cgctgccccg tctggtgatc ctcagcagat ttttttgggt 1020

acgccgccgg ggccgttggc tgacgggtct gtggtgttgc gtcttcgcgg gcaggctttg 1080

tcgggtggta aacggtttgc gtggacggag ttttcgattc ctgacgagtc tgatccggat 1140

gatgtgtcgc ggcagtggcg gaagttggcg ggtgacacta atccggcgtt ggggcgccgc 1200

ctgaatttcg ggacagtctc ggatgagcat gagtcgatgt ctgctgccgg gtttgctcgg 1260

gagcggcttg gctggtggga tcgtggccag tctgcttcgt ctgtgattcc ggcggataag 1320

tgggttcagt cggctgtggt tgaggcggct ctggttggcg ggaaggtttt tggtgtctcg 1380

ttttctcgct cgggggatcg tgtcgcgttg gctggtgctg gtaaaacgga ttctggtgtg 1440

catgttgagg ttattgatgg cctgtctggg acgattgttg atggtgtggg ccagctggct 1500

gattggttgg cgttgcgttg gggtgacact gaaaaggtta tggttgcagg gtctggtgcg 1560

gtgttgttgc agaaggcttt gacggatcgt ggtgttccgg gtcgtggcgt gattgtggct 1620

gatactgggg tgtatgtgga ggcgtgtcaa gccttcctgg agggtgtcag gtctgggagc 1680

gtgtctcatc ctcgtgccga ttcgaggcgt gacatgttgg atattgctgt gaggtcggct 1740

gtgcagaaga agaagggttc tgcgtggggt tggggttcct cgtttaagga tggttctgag 1800

gttcctttgg aggctgtgtc tttggcgtat cttggtgcga agatggcgaa agcgaagcgg 1860

cgtgaacggt ctggtaggaa gcgggtgtct gtggtatgaa ctcggatgag ttggctctga 1920

ttgagggcat gtacgatcgt attcaagggt tgtcttcgtg gcattgccgt attgagggct 1980

actatgaggg ctctaatcgg gtgcgtgatt tgggggttgc tattccttcg gagttgcagc 2040

gggtgcagac ggtggtgtca tggcctggga ttgcggtgga tgctttggag gagcgtctgg 2100

attggcttgg ctggactaat ggtgacggct acggtttgga tggtgtgtat gctgcgaatc 2160

ggcttgctac ggcgtcgtgt gatgttcacc ttgatgcact gatttttggg ttgtcgtttg 2220

tggcgatcat tccccaagag gatgggtcgg tgttggttcg tcctcagtcg ccgaagaatt 2280

gtactggccg gttttctgcc gatgggtctt gtttggatgc tggccttgtg gtgcagcaga 2340

cgtgtgatcc tgaggttgtt gaggcggagt tgttgcttcc tgatgtgatt gttcaggtgg 2400

agcggcgggg ttcgcgtgag tgggttgaga cgggccgtat cgagaatgtg ttgggtgcgg 2460

ttccgttggt gcctgttgtg aatcgtcgcc gtacttctag gattgatggc cgttcggaga 2520

ttacgaggtc tattagggct tacacggatg aggctgttcg cacactgttg gggcagtctg 2580

tgaatcgtga tttttatgcg tatcctcagc gttgggtgac tggcgtgagc gcggatgagt 2640

tttcgcagcc gggttgggtt ctgtcgatgg cttctgtgtg ggctgtggat aaggatgatg 2700

atggtgacac tccgaatgtg gggtcgtttc ctgtgaattc tcctacaccg tattctgatc 2760

agatgcgttt gttggcgcag ttgactgcgg gtgaggcggc tgttccggaa cgctatttcg 2820

ggtttatcac ttctaacccg ccttctgggg aggctttggc tgcggaggag tctcggcttg 2880

tgaagcgtgc tgaacgcagg cagacgtcgt ttggtcaggg ctggctgtcg gttggtttcc 2940

tggctgcccg ggcgttggat tcgagtgttg atgaggccgc gttttttggt gatgttggtt 3000

tgcgttggcg tgatgcgtcg acgccgactc gggcggctac ggctgatgct gtgacgaagc 3060

ttgtgggtgc tggtattttg cctgctgatt ctcggacggt gttggagatg ttgggtttgg 3120

atgatgtgca ggttgaggct gtgatgcgtc atcgtgccga gtcttcggat ccgttggcgg 3180

cactggctgg ggctatttcc cgtcaaacta acgaggtttg ataggcgatg gcttcgggtg 3240

ctgtgtcgag gcttgctgcg actgagtatc agcgtgaggc tgtcaggttt gctgggaagt 3300

atgcgggcta ttatgccgag ttgggtcgtt tgtggcgtgc cggcaggatg agtgacacgc 3360

agtatgtgcg tttgtgtgtg gagttggagc gtgccggcca tgacggttca gcagctatgg 3420

cgggcaaatt cgtttcagat tttcgccggt tgaatggtgt cgatcctggt ttgatcgtgt 3480

atgacgagtt tgatgctgcg gcggctttgg ctaggtcgtt ttcgactatg aagattatga 3540

atagtgaccc ggatagggcg aatgatacga ttgatgcgat ggctgcgggt gttaatcggg 3600

ctgttatgaa tgctggtcgt gacacggttg agtggtcggc gggtgcgcag ggtaggtcgt 3660

ggcgtcgggt gactgatggt gatccgtgtg ctttttgtgc catgttggct acgaggtcgg 3720

attatacgac taaagagcgg gcgcttacta ctggtcatac gcggcgtcat aagcgtgccg 3780

gtaggcgtcc gtttggttcg aagtatcatg atcattgtgg ttgtacggtg gttgaggttg 3840

ttggtccttg ggaaccgaat agggctgatg ccgagtatca gaggacgtat gagaaggctc 3900

gtgagtgggt tgatgatcat gggttgcagc agtcgtctgg caatattttg aaggctatgc 3960

gtactgttgg tggcatgaga taatttgatg tggtttccgg ttgtgtgccg ccggttatcg 4020

gtgcacaggg ttgtctcccg cacgggggtc aacaatgttg tgttgttttc cgcaaggagt 4080

gtagggttag gctatggccg atcagagtat tgaggaacag aatgttgaca atgatgttgt 4140

ggagtccgga aaggataacg gcattgttga tacagtaaaa gacgatggcg ggcaggaggt 4200

agccgacaat cagttgaaga atgaaggcga gggtaaatcg ccggggactg attggaaggc 4260

ggaggcccgt aagtgggagt ctcgtgctaa aagtaatttc gccgagttgg agaagcttcg 4320

tacatcgagt gacgattctg gatctactat tgatgagctt cgccgcaaga atgaggaact 4380

cgaagaccgg attaacgggt ttgttcttga gggtgtgaag cgcgaggtgg ctgccgagtg 4440

tggcctgtcg ggtgatgcga tcgcttttct tcacggtagc gataaggagt cgcttgccga 4500

gtctgctaag gctttgaagg gtttgatcga ccatagtagt ggtggtggcg cgggtgtgcg 4560

ccgtcttgcg gggagtgccc ccgttgatga tgttaaacga cgtgagggtg tcgcgtttgt 4620

ggatgctctt gtcaataatt ctaggagatg atttatcatg gctgacgatt ttctttctgc 4680

agggaagctt gagcttcctg gttctatgat tggtgcggtt cgtgaccgtg ctatcgattc 4740

tggtgttctt gctaaactgt caccggagca gccgactatt ttcgggcctg ttaagggcgc 4800

cgtttttagt ggtgttccgc gcgctaagat tgttggcgag ggcgatgtta agccttccgc 4860

tagcgttgat gtttctgcgt ttactgcgca gcctatcaag gttgtgactc agcagcgtgt 4920

ctcggacgag tttatgtggg ctgacgccga ttaccgtctg ggtgtgcttc aggatctgat 4980

ttccccggcc ctgggtgctt ctattggtcg cgccgttgat cttattgctt tccatggtat 5040

tgatcctgct acgggtaagc ctgctgcggc tgtcaaggtg tcgctggata agacgaataa 5100

gacggttgat gccaccgatt ccgctacggc tgatcttgtt aaggctgttg gtctgattgc 5160

tggtgctggt ttgcaggttc ctaacggtgt tgctttggat ccggcgttct cgtttgctct 5220

gtcaactgag gtgtatccga agggttcgcc gcttgccggt cagccaatgt atcctgccgc 5280

cgggttcgcc ggcctggata attggcgcgg cctaaatgtt ggttcttctt cgactgtttc 5340

tggtgccccg gagatgtcgc ctgcttctgg tgttaaggct attgttggtg atttctctcg 5400

tgtccattgg gggttccagc gtaacttccc gattgagctg atcgagtatg gtgacccgga 5460

tcagacgggg cgtgacttga agggccataa tgaggttatg gttcgtgccg aggctgtgct 5520

gtatgttgcg attgagtcgc ttgattcgtt tgctgtcgtg aaggagaagg ctgccccgaa 5580

gcctaatccg ccggccggta actgattcat ttgttgcgat aatgtttatg ctgtgtgcag 5640

ggggtggtgt tgatgggtat cattttgaag cctgaggata ttgagccttt cgccgatatt 5700

cctagagaga agcttgaggc gatgattgcc gatgtggagg ctgtggctgt cagtgtcgcc 5760

ccctgtatcg ctaaaccgga tttcaaatat agggatgccg ctaaggctat tctgcgtagg 5820

gctttgttgc gctggaatga tactggcgtg tcgggtcagg tgcagtatga gtctgcgggc 5880

ccgtttgctc agactacacg gtcgaatact cctacgaatt tgttgtggcc ttctgagatt 5940

gccgcgttga agaagttgtg tgagggtgat agtggggctg gtaaggcgtt cactattaca 6000

ccgaccatga ggagtagtgt gaatcattct gaggtgtgtt ccacggtgtg gggtgagggt 6060

tgctcgtgcg ggtcgaatat taacggctat gctggcccgt tgtgggagat atgatatgac 6120

cggttttcct tacggtgaaa cggttgtgat gcttcagccg actgttcgtg tcgatgatct 6180

tggtgacaag gtggaggatt ggtctaagcc tgtcgagact gtgtaccata acgtggccat 6240

ctatgcttcc gtttcgcagg aggatgaggc cgcggggcgt gactcggatt atgagcattg 6300

gacactgctg ttcaagcagc ctgtcaaggc tgctggttat cggtgtcgtt ggcgtattcg 6360

gggtgttgtg tgggaggctg acgggtctcc tatggtgtgg catcatccga tgtctggctg 6420

ggatgctggt acgcaggtta atgtgaagcg taagaagggc tgatgggttg tggcacgtga 6480

tgttgatgtg aagctgaact tgccgggtat tcgtgaggtg ttgaagtctt ctggggtgca 6540

gggcatgttg gctgagcgtg gtgagcgtgt caagcgtgcg gcctcggcga atgtgggcgg 6600

taacgcttac gatagggccc agtatcgtgc cgggttgtcg tctgaggtgc aggttcaccg 6660

tgttgaggct gtggcgcgta ttggcaccac ctataagggt ggtaaaagga ttgaggctaa 6720

gcatggcacg ttggcgaggt cgattggggc tgcgtcgtga tcgtttacgg tgatcctcga 6780

atatgggcta aacgtgtgtt ggcggatgat ggttggctgt ctgatgtacc gtgcacgggt 6840

actgtgccgg atacatttga gggtgatctg atttggttgg cgttggatgg tggcccggag 6900

ttgcatgttc gtgagcgtgt ttttttgcgt gtgaatgtgt tttcggatac gccggatcgt 6960

gctatgtctt tggctcgccg ggttgaggct gtgctggctg atggtgtgga tggtgatccg 7020

gtggtgtttt gcaggcgttc gactgggcct gatttgctgg tggatggtgc acgttttgat 7080

gtgtattcgc tttttgagct gatatgtagg cctgcggagt ctgaataagc ttattgtttt 7140

tgttttaatg taattgtttg atatttaatg ggggttgtga tggctgctac acgtaaagcg 7200

tctaatgttc gttcagcggt tactggcgac gtttatattg gtgacgcgca cgcgggtgat 7260

tctattaagg gtgtggaggc ggttccttcc gggcttacag ctttggggta tctgtctgat 7320

gacgggttta agattaagcc tgagcgtaaa acggatgatt tgaaggcttg gcagaatgcg 7380

gatgttgttc gcactgtggc tacggagtcg tctatcgaga tttctttcca gctgattgag 7440

tcgaagaagg aggttatcga actgttttgg cagtcgaagg ttactgccgg atctgattcg 7500

ggttcgttcg atatttctcc tggtgccaca acaggtgttc acgccctgtt gatggatatt 7560

gttgatggcg atcaggttat tcgctactat ttccctgagg ttgagctcat tgatcgtgac 7620

gagattaagg gcaagaatgg cgaagtgtac gggtatggtg tgacgttgaa ggcgtatcct 7680

gcccagatta ataagactgg taatgcggtg tcgggtcggg ggtggatgac ggctttaaaa 7740

gctgatactc ctccgactcc tccgccggcc ccggttcctc cgaagcctca gccggatccg 7800

aatccgccgt ccggtaactg atacacgatt ttaggggatt gttaatagat gagtgacact 7860

ggtttcacgt tgaagattgg tgatcgtagc tgggtgttgg cggatgcgga ggagacggct 7920

caggctgttc ctgcccgcgt tttccgtcgt gccgccagga ttgcccagtc gggggagtct 7980

gcggatttcg cccaggttga ggtgatgttt tctatgttgg aggctgccgc cccagctgac 8040

gcggtggagg ccctggaggg gcttcctatg gttcgtgtgg cggaggtttt ccgtgagtgg 8100

atggaataca agcctgacgg taagggtgcc tcgctggggg aatagtttgg ctccacggcc 8160

tgattgatga ttatcgtggg gccatcgaat acgatttccg caccaagttt ggtgtttctg 8220

tttatagtgt tggtggcccg cagatgtgtt ggggtgaggc tgtccggctg gctggcgtgt 8280

tgtgtaccga tacgtctagc cagttggcgg cccaccttaa tggttggcag cgcccgtttg 8340

agtggtgcga gtgggctgtg ttggacatgt tggatcatta caggtctgct aatagtgagg 8400

ggcagccgga gcctgtggcg aggccgactg atgagcgtcg ggcaaggttt acgtctgggc 8460

aggtggacga tattttggcg cgtgttcgtg ccggtggcgg ggtgtctcgc gagattgata 8520

ttatggggtg aatagtgtat gtctggtgag attgcttccg catatgtgtc gttgtatacg 8580

aagatgcctg gccttaaaag tgatgttggt aaacagttgt cgggtgttat gcctgctgag 8640

gggcagcgtt cgggtagcct gtttgctaaa ggcatgaagt tggcgcttgg tggtgcggcg 8700

atgatgggtg ccatcaatgt tgctaagaag ggcctcaagt ctatctatga tgtgactatt 8760

ggtggcggta ttgctcgcgc tatggctatt gatgaggctc aggctaaact gactggtttg 8820

ggtcacacgt cttctgatac gtcttcgatt atgaattcgg ctattgaggc tgtgactggt 8880

acgtcgtatg cgttggggga tgcggcgtct acggcggcgg cgttgtctgc ttcgggtgtg 8940

aagtctggcg gtcagatgac ggatgtgttg aagactgtcg cggatgtgtc ttatatttcg 9000

ggtaagtcgt ttcaggatac gggcgctatt tttacgtctg tgatggctcg cggtaagttg 9060

cagggcgatg acatgttgca gcttacgatg gctggtgttc ctgtgctgtc tttgcttgcc 9120

aggcagacgg gtaaaacctc ggctgaggtt tcgcagatgg tgtcgaaggg gcagattgat 9180

tttgccacgt ttgcggctgc gatgaagctt ggcatgggtg gtgctgcgca ggcgtctggt 9240

aagacgtttg agggcgctat gaagaatgtt aagggcgctt tgggctattt gggtgctacg 9300

gctatggcgc cgtttcttaa cggcctgcgg cagatttttg ttgcgttgaa tccggttatt 9360

aagtctatca cggattctgt gaagccgatg tttgctgccg tcgatgctgg tatccagcgg 9420

atgatgccgt ctattttggc gtggattaac cgtatgccgg ctatgatcac gagaatgaat 9480

gcacagatgc gcgccaaggt ggagcagttg aagggcattt ttgcgagaat gcatttgcct 9540

gttcctaaag tgaatttggg tgccatgttt gctggcggca ccgcagtgtt tggtattgtt 9600

gctgcgggtg tggggaagct tgttgcaggg tttgctccgt tggcggttgc gttgaagaat 9660

ctgttgccgt cgtttggtgc tttgaggggt gccgccgggg ggcttggtgg cgtgtttcgc 9720

gccctgggtg gccctgtcgg gattgtgatc ggcttgtttg cggcaatgtt tgccacgaac 9780

gcccagttcc gtgccgctgt tatgcagctg gtggctgtgg ttggtcaggc gttgggccag 9840

attatggcag ctgtgcagcc gctgtttggt ttggttgctg gcgtggttgc caggttggcg 9900

ccggtgttcg gccagattat cggtatggtt gctggtttgg ctgcccggct ggtgcctgtt 9960

attggtatgc ttattgcccg gctggttcct gttatcaccc agattattgg tatggtaacc 10020

caggttgctg ccatgttgtt gcctatgctg atgccggtta ttcaggctgt tgttgctgtg 10080

atacggcagg ttattggtgt cattatgcag ttgatacctg ttttgatgcc ggttgtgcag 10140

cagattttgg gtgctgtcat gtctgttttg ccgccgattg ttggtttgat acggtcgctg 10200

ataccggtga tcatgtcgat tatgcgtgtg gtggtgcagg ttgttggtgc tgtgctacag 10260

gtggtggccc gtattattcc ggttgttatg ccgatttatg tttcggtgat tggattcatt 10320

gccaagattt atgctgcggt tatcgttttt gaggctaagg ttattggcgc tattcttcgt 10380

actattacgt ggattgtgaa tcattcagtg tctggcgtga ggtctatggg cacggccatc 10440

cagaatggct ggaatcatat taaatcgttt acgtctgcgt ttattaacgg ttttaagtcg 10500

atcatttctg gcggcgtgaa cgcggttgtg gggtttttta cgcggcttgg tttgtcggtt 10560

gcttcccatg tgaggtccgg ttttaacgct gcgaggggtg ctgtttcttc cgccatgaat 10620

gctattcgga gtgttgtgtc ttcggtggcg tctgctgttg gcgggttttt cagttcgatg 10680

gcgtctcgtg ttcggaatgg tgctgtgcgc gggtttaatg gtgcccggag tgcggcttct 10740

tctgctatgc atgctatggg gtccgctgtg tctagtggtg tgcatggtgt gctgggtttt 10800

ttccggaatt tgcctgacaa tattcggcgt gcgcttggta atatggggtc cctgttggtg 10860

tcggctggcc gtgatgtggt gtccggttta ggtaatggta tcaagaatgc tttgagtggc 10920

ctgttggata cggtgcgtaa tatgggttct caggttgcta atgcggcgaa gtcggtgttg 10980

ggtattcatt ccccgtctcg ggtgtttcgt gacgaggttg gccggcaggt tgttgccggt 11040

ttggctgagg gtattactgg taatgctggt ttggcgttgg atgcgatgtc gggtgtggct 11100

gggaggctgc ctgatgcggt tgatgcccgg tttggtgtgc gatcgtctgt gggttcgttt 11160

accccgtatg gcaggtatca gcgcatgaat gataagagtg ttgtggtgaa tgtgaatggg 11220

cctacttatg gggatcctgc cgagtttgcg aagcggattg agcggcagca gcgtgacgct 11280

ttgaacgcgt tggcttacgt gtgattttgg gggtgtggtg catgtttatt cctgacccgt 11340

ctgatcgttc tggtttgact gtgacttggt ctatgttgcc gttgattggt aatgatccgg 11400

agcgtgtgct tcatttgacg gattatacgg ggtcgtctcc gataatgttg ttgaatgatt 11460

cgttgcgcgg tttgggtgtt cctgaggtgg agcatttttc tcaaactcat gttggggtgc 11520

atggctcgga gtggcgcggg tttaatgtga agcctcgcga ggtgacgcta ccggtgttgg 11580

tgtcgggtgt tggcccggat ccggtgggcg gttttcgtga cggttttttg aaggcgtatg 11640

acgagttgtg gtctgctttt cctcctggcg aggtggggga gttgtctgtg aagactcctg 11700

ccggtcgtga gcgtgtgttg aagtgccggt ttgattcggt ggatgacacg tttacggtgg 11760

atccggtgaa caggggttat gcgcgttatc tgttgcattt gacggcttat gacccgtttt 11820

ggtatgggga tgagcagaag tttcgtttca gtaacgctaa gttgcaggat tggttgggtg 11880

gcggccctgt cgacggtaag ggtaccgcgt ttccggtggt gttgacgcct ggtgttggtt 11940

cgggttggga taatctgtct aataagggtg atgtgcctgc gtggcctgtg attcgtgttg 12000

aggggccgtt gtcgtcgtgg tctgtgcaga ttgatggttt gcgtgtgtcc tcggattggc 12060

cggtggagga gtatgattgg atcactattg atacggatcc tcgtaagcag tctgcgttgt 12120

tggacgggtt tgaggatgtg atggatcgtt tgaaggagtg ggagtttgcg cctatcccgc 12180

ctggcggttc tcggagtgtg aatattgaga tggttggttt gggtgccatt gttgtgtcgg 12240

tgcagtacag gtttttgagg gcttggtgaa tagttgatgg ctggttttgt tccgcatgta 12300

acattgttta caccggatta tcgccgtgtg gcgcctatca atttttttga gtcgttgaag 12360

ttgtcgttga agtggaatgg tttgtccact ttggagttgg tggtgtctgg tgatcattct 12420

aggcttgacg ggttgactag gccgggtgcg cggcttgtgg ttgattatgg tggtggccag 12480

attttttctg ggcctgtgcg tcgggtgcat ggtgtgggtc cgtggcgttc ttcgcgtgtg 12540

actatcacgt gtgaggatga tattcgtctg ttgtggcgta tgttgatgtg gcctgtgaat 12600

tatcgtcctg gtatggttgg tatggagtgg cgtgcggatc gggattatgc ccattattcg 12660

ggtgcggcgg agtcggtggc taagcgggtg ttgggggata atgcttggcg ttttccgtct 12720

ggtttgttta tgaacgatga tgagagtcgt ggccgctata ttaaggattt tcaggtgcgg 12780

tttcacgtgt ttgccgataa gttgttgccg gtgttgtcgt gggctcggat gactgtcacg 12840

gtgaaccagt ttgagaatgc gaagtttgat cagcgtggtt tggtgtttga ttgtgtgcct 12900

gctgtgaccc ggaaacatgt gttgactgcc gagtcgggtt cgattgtgtc gtgggagtat 12960

gtgcgtgacg ccccgaaggc gacatctgtg gtggttggtg gccgtggcga gggtaaggat 13020

cggctgtttt gtgaggatgt tgattcggcg gccgaggatg attggtttga tcgtgtcgag 13080

gtgtttaagg atgcccgtaa cacggattcc gagaaggtgt ctctcttcga tgaggctgag 13140

cgggtgttgt ccgagtcggg ggctacgtcg gggtttaaga ttgagttggc tgagtcggat 13200

gtgttgcggt ttggtcccgg caatctgatg cctggggatt tgatctatgt ggatgtgggt 13260

tctgggccta ttgcggagat tgtgcggcag attgatgtgg agtgtgtatc gcctggtgat 13320

ggttggacga aggtgactcc ggttgcgggg gattatgagg ataatccgtc ggccctgttg 13380

gctcgccgtg tggctggttt ggctgcgggt gtgcgggatt tgcaaaagtt ttagtaagtg 13440

attggggttt gttgtgggta ttgtgtgtaa agggtttgat ggtgtgttga ccgagtatga 13500

ttgggctcaa atgtctggtc tgatgggtaa tatgccgtct gtgaaggggc ctgacgattt 13560

tcgtgtcggc acgacgattc agggttctac ggtgttgtgt gagatcctgc cggggcaggc 13620

ttgggctcac ggggtgatgt gcacgtcgaa tagtgttgag acggtgacgg gtcagcttcc 13680

gggcccgggt gagactcgat acgactatgt ggtgttgtct cgggattggc aggagaatac 13740

ggccaagttg gagattgttc ccggtgggcg tgcggagcgt gccagggatg tgttgagggc 13800

tgagcctggc gtgtttcatc agcagctact ggcgactttg gtgttgtcgt ctaacgggtt 13860

gcagcagcag ttggataggc gtgctgtggc ggctagggtt gcgtttgggg agtctgctgc 13920

gtgtgatcct acccctgtgg agggtgaccg tgtgatggtt ccttcggggg ctgtgtgggc 13980

taaccatgcc ggcgagtgga tgttgttgtc tcccaggatt gagacgggtt cgaagtcgat 14040

catgtttggt ggttctgctg tgtatgctta cacgatcccg tttgagcgcc agttcagtag 14100

tccgcctgtt gtggtggcgt ctatggctac ggcggctggg ggcacggcac agattgatgt 14160

gaaagcctac aatgtgactg cccaaaattt tagtttggcg tttattacga atgatggttc 14220

gaagccgaat ggtgtgcctg cggtggcgaa ttggattgct gtcggcgtgt gactgcacgg 14280

gtgttgtggc ggatggtgtg atgttggggg gctgtggtgt cgtggtttac tcctgcactg 14340

gtggcctcta tttgtaccgc gttggccacg gttttgggtt ctgttcaggc tgtcacatcc 14400

cggtctagga agcgtttacg caggctgtcg gctcaggtgg atgcgatgga agagtatacg 14460

tggggtgtgc ggcgcgaggt gcgaaggttt aacgccgggc ttcctgatga tgtggagccg 14520

atgcatcttc ctgatttgcc cgagtttttg aaagatactg ttgatggtgg aggtgagtag 14580

ggttgaggga gttggaggag gagaagcggc agcgccgcaa ttttgagaag gcttcactgg 14640

tgttgttgtt tttgtcgctt gtgttgttgg cggtggttgc tgcgggtgct ttgcgtttcg 14700

gggctgtatc ctctgagcgg gattcggagc aggcgagggc ccagtcgaat ggtacggctg 14760

ccaggggttt ggctgcccgt gtgaagcagg cgtgtgcttc gggtggggtg gagtctgtgc 14820

gtcttcaccg ttctggtttg tgtgtggatg ctgtgcgtgt tgagcagcgt gttcagggtg 14880

tgccgggtcc tgccggtgag cgcggcccgc aaggcccttc aggtcctgcc ggccgggatg 14940

gtgttaatgg ttcggctggg ctggttggcc ctgttggtcc gcaaggttct ccgggtttga 15000

atggtgtgaa aggtcctgac ggcttgcctg gcgctaacgg ttcggatggc cgtgatggtg 15060

ttccaggtcg tgcaggtgct gacggtgtga acggcgttga cggcgctgat ggtcgggatg 15120

gttctgccgg tgagcgcggc ccgcaaggcc cttcaggtcc tgccggcccg caaggtgcac 15180

agggtgaacg gggtgagcgt ggtcccgccg gtgcgaatgg atcggatggc catgatggta 15240

aggatgggcg ctcggtggtg tctgtgtact gttccggggg ccgcctggtt gtgaaatata 15300

gtgacggtgt ggcttccacg atatcgggtt cggcggcctg ccagggtgtg aaaccgtcgc 15360

ctctagtgac tatatcatcc cacaaataga aaggagtggc tgtgatggtg gtgtttggtg 15420

gtggtgtgtt gtgagatata ttcctgcggc gcatcattct gccggctcga atagtccggt 15480

gaatagggtt gtgattcatg cgacgtgccc ggatgtgggg tttccgtccg cctcgcgtaa 15540

aggacgggct gtgtccacgg caaactattt cgcttcccca tcgtctggtg gttcggcgca 15600

ttatgtgtgt gatattgggg agacggtgca atgcttgtcg gagtctacga ttgggtggca 15660

tgccccgccg aatccgcata gtttgggtat agagatttgc gcggatgggg gttcgcacgc 15720

ctcgttccgg gtgccggggc atgcttacac tcgtgagcag tggctggatc ctcgcgtgtg 15780

gcctgcggtt gagcgtgccg ccatcctgtg tagacgtttg tgtgacaagc atggtgttcc 15840

gaaaaggaaa ctgtctgtgg ccgatttgaa ggccggtaaa cggggtgttt gcgggcatgt 15900

ggatgttacg gatgcgtggc atcagtcgga tcatgacgat ccggggccgt ggtttccgtg 15960

ggacaaattt atggctgtgg ttaatggcca cggcggcggt tcaagtagtg aggagttgag 16020

tatggctgat gtacaagcgt tacataatca gattaaacag ttgtcggcac aggtggccca 16080

gtcggtgaat aagctgcatc acgatgttgg tgtggttcag gttcagaatg gtgatttggg 16140

taaacgtgtt gatgccttgt cgtgggtgaa gaatcctgtg acggggaagc tgtggcgcac 16200

taaggatgct ttgtggagtg tctggtatta cgtgttggag tgtcgtagcc gtcttgacag 16260

gctcgagtct gctgtcaacg atttgaaaaa gtgatggtgg tttgttgtgg gtaaacagtt 16320

ttggttaggt ttgctggagc gtgccctgaa aacttttgtt caaacgtttg ttgccgtgtt 16380

gggggttact gcgggtgtca cctatactgc ggagtcgttt cgtggtttgc cgtgggaatc 16440

cgcgctgatc acggcaacgg ttgctgctgt cctgtcggtt gctacctcgt ttggtagccc 16500

gtcgtttgtg gccggcaagc ccggcaagca gccccaggtg gatgcgggtt tggttccacc 16560

ggatgatggg ggcttggttg agccgcatat ggtggatgtg tcggatcctg gcatgatcga 16620

gccgacggat gatgcggatc ttgccggcta tgagcctcgg cgtgcagccg agtcggaggt 16680

tggcacggta gagtctactg ttgcataatt gaatatagat gtgtgcccca gcggtgctgc 16740

cacgattgtg tggtggcggc tgctggggca ctatttttgt atatgcggtg tggctatgat 16800

tcgttgctgt cgatggtgtc ttcgagcatc tgatacaggt ggaggcaggt agagatagtt 16860

tcgctggcct gatcgagaac gttccggccg ataacgtttt tgtggttgtc gcggtggcgg 16920

atgatagccc acatgatctc gtcggctgcc gcctgtaata gtttggcctg gtatgcgatt 16980

ccggcgagcc agtctagtgc ttcctggctt gtataggggc tctggtcctc gctgttgccg 17040

cgggtgttgc tgttgtttgt ggggtgtcct gcactgtcgc atagccacag gatttcgctg 17100

cactcgtcta gcgtgtcttg gtcgatagcg agatcgtcga ggctgacatt gttgacggta 17160

aggttcacgt tgtcgaggga gatgggtaca ccgtactggt tttcgacact gtcaacaatg 17220

ttttccagct gttgcatgtt ggtgggctgt tgttggacga tacggtgtat cgctgtgttg 17280

agggtggtgt aggtgatgtt gtgtgtgttg tccatggttt ttatgccatt ccttcgttat 17340

cgtctggcat gtagtatgtg ctgtttgcgt actcggttaa cgtcatcagt gtttggtctg 17400

cccactgttt cacggtttgc cgggtgactc cgagtcgttg ggcggctgtg gcgtaggttt 17460

gatcataccc gtatacttcc cggaatgctg ccaacctagc taggtgtttc ctctgtttgg 17520

atggttcaca ggtgagggtg tagtcgtcga tggctagctg tagatcgatc atggagacga 17580

tgttgttgcc gtggtgttgt ggcgcggttg gtgggggtgg cattcctggc tccacggagg 17640

gtttccaggg gccgccgttc cagatccatt gggcagcttg gatgatgtcg gcggtggtgt 17700

aggttcggtt cactggtcac cccctgaaca ggtcgttggt gttgttggtg tcgaatcgtc 17760

cgacgcagtg gcagtagtcg tacatgagtt taataatgtg ttggtggtct cccaaatagg 17820

tgtttccgct gatgctgtat gtggctgtgc cgtctttcgc gatggtgtat ttggcggtga 17880

tggtttcggg gttttcggtg tcggtgatga ttgctgtggt ggtggcgcct actgtttgga 17940

gtatggtggt ttgggttccg tcgtcgatgg tggttttaac catggtgtgt gttttccctt 18000

ttgttagttg cttgtttggt tgtcggctag atgaataata tcgggtaaag gtttcggctg 18060

gtctaggtgt tgtatggttt tgttggctag ccgtttggct accctgtaac acattttggt 18120

gtagtgtttg ttgtctaggt tgtggtattg ttcccgcacc gcaatatata gcagggagtc 18180

ttggtacagg tcgtctgcac tgattgcggg gtagtgtgcg gctgttttgg tgcatgcccg 18240

gttgagtgtg cgaagatgat ggtctgtggc ccacacccac gatgcggtgg tggccaggtc 18300

ggcttttgtt ggtcgtctgc tcatggcact atttcatctc gctatctgat agttgtttgg 18360

tgttttgttg tggatagtgt agcacactag tcctgggtgg ccggtggtgc ctgtgcggtg 18420

acggaaccat gtggattcgc cttccatgga tgggcattgg atgaaggtgc gttgtccttg 18480

ctcggagatt tctaggtggt gccggtgccc ggccatgaga atattagata cggtgccgtt 18540

gtggaattct tggccgcgcc accaatcata gtgtttaccg gtgcgccatt ggtgcccgtg 18600

ggcgtgcagt atccgtgtgc ctgccacatc aacggtggtg gtcatttcgt ctcggctggg 18660

gaagtggaag tgtaggttgg ggtattggtt attgagctgg taggcttctg cgatggcccg 18720

gcagcagtcc acgtcgaatg agtcatcgta ggtggtgact cctttaccga agcgcacggc 18780

ttcaccatgg ttgccgggga tggatgtgat ggtcacattt ttgcagtggt cgaattggtg 18840

gatgagttgc atcatggcca tgcgggtgag cctgatttgt tcggtgaggg gtgtttgtgt 18900

tcgccaggcg ttgttgcctc cttgtgacac gtatccttcg atcatgtcgc cgaggaaggc 18960

gatgtggact cgttcgggtt tgcctgcttg ttgccagcag tgttttgcga ctatgaggga 19020

gtgtaggtag ttgtcggcga agtgtgctgt ttctccgccg gggatgcctt tgccgatttg 19080

gaagtctcct gccccgatga cgaaggctgc ggtgctgtag tcggtgtggg tgtcttgttc 19140

gggttttggg ggtgtccatt cggctagttt atcgacgagt tcgtctaccg ggtaggggtt 19200

tgttgcgggt tggtggtcga tgattttttg tatggatcgg cctgtttctc cgttggggag 19260

tgtccattcg gagatgcgtg tgcggcgcac ggtgccgttg gctagattgt cgtcgatggt 19320

gtcgatggcg ttgtcgtggt tggctagctg tgtgagtagc cggtcaatat tgtctatcac 19380

tgggtatcct cctcttgcgg ggtggtgctg gcttgtttgc ggcgatagtc tttaataacg 19440

gtggcggaga tggggtatcc tgcctgggtg agctgttttg ctagccatga ggcggggata 19500

gacctgtcgg cgagcacgtc ggcggctttg ttgccgtagc gttgaataag ggtttcagtt 19560

ttggttgcca tgatgtccta tcggttgtgt ggtgggctgc catcctgtgc ggcagtcgcc 19620

gtcgtgtcct ggtttgcgtg tgcaccacga tacggttccg tctgtgtggt tgagtgtttt 19680

accgcacatg acgtttcgga gatgctccgg cagctggtca tcctggttgc tggtttgtgt 19740

gtcgaagagt gttttctggt tggtgaaatg ttctgacacg gtgccgttat gcacgggtag 19800

tatccatgtt ttccattgtt gttgtagcct ggtgttccag tggaattgtt tggcggcgtt 19860

ttcggcctgt tttaaggttt tgtggtagcc gactagtatg cgttgatgct gctggtctgg 19920

agggtttggg cctcgccagt attgtgccgc cacggcgtag cggttgctgt ctgtgaaggc 19980

gtcccagcag tattcgataa tgtgttgcaa catactgtct ggcaggctgt cagggttgat 20040

gttgatgttt tgggtgataa tgtcacggat ggcttgccgg tttttggtgg tgggtttgaa 20100

cgagatgctc acgatagtac cggctggtcg tcttgcatga actggttgaa ggtgttgttc 20160

ccggcgtgtt gggcttgtgt tatttgttgg tcggtccagt ctgggtgttg ctgtttcaga 20220

tagtgccagt ggcacgcatt gtaggtttcg tcttgtagcc gtgtgagatg gttttcggtg 20280

atgatttgtt tccacatggc ccatgacacg tcgagccggt cgaggatttc gagggctggg 20340

atgttgaatt ggttcaggaa gaggatttcg tgggtgtagt agtttttctc gtaggcgtcc 20400

catccgcttc ggtgcctgtt gggctggttt ttggggtagg cttcccggca tactttgtgt 20460

aaacgcttgg ccatgtcgtc gggtagttta atgtcggggt tggcgcggat catggatcgc 20520

atcccatcat aggtggtgcc ccaggtgtgc atgatgtagg tggggtcttc tccgtcggcc 20580

catttttctg cacagatggc gaggcggata cgcctcctgg cagcttggct ggtgttgcgc 20640

cggttgggga ttgggcacgt gtcgagggga tccatgatgt tttagtgtac ctttctggtt 20700

tcgtgttgtt gacaggtttt actgtagcac agtgtctagt gcgtgtgtca accctgtttt 20760

tccggcttga aggtaggtgt ctgtgacatc ccctagggtg aggggcacgt gcacagcttg 20820

ggggagtgcc gcctggaggg tttgggccat ctggtcgcct gcggggtctg ggtctgacca 20880

gatgtagatg tggtcgtagc cttcaaaaaa tttggtccaa aaaatttgcc acgaggttgc 20940

gccgggtagg gcgacggccg accatccgca ttgttcgagg atcatggagt cgaattcgcc 21000

ttcgcaaatg tgcatttcgg ctgccgggtt ggccatggcg gccatgttgt agatggagcc 21060

tgtgtctcct gccggggtta ggtatttggg gtggttgtgg gttttgcagt cgtgcgggag 21120

tgagcagcgg aaacgcattt ttcttatttc ggctgggccg ccccaaacgg ggtacatgta 21180

tgggatggtg atgcactggt tgtagttttc gtggcctggg atggggtcat tgtcgatgta 21240

tccaaggtgg tggtagcggg ctgtttcttc gctgatgcct cttgctgaga gcaggtcgag 21300

tatgttttcg aggtgggttt cgtagcgggc tgaggctttc tggattcggc ggcgttccgc 21360

aatgttgtat gggcgtatgc tgtcgtacat ttgggttttc ttcttctaat cgttgttgta 21420

gcttggcgag tccgcctccg acaccgcatg tgtggcagta ccagacgccc ttgtcgaggt 21480

tgatgctcat ggagggctgg tggtcgtcgt ggaacgggca gagtatgtgt tgctcgttcc 21540

tggacggatt gtaccgtatc tgataatggt cgaggaggcg gcaggtgtca gaggtgtggg 21600

aggagctcgt tgagggttga taccacatag gcttcactcc atggcttgtt gcgctgtttc 21660

atcactacga gtccgatggt ggaattgttt tgtttgtttc ggtgtgtttc gtagttgcgt 21720

gcctcccggc tggcttgttt cacgaattgg gctaggtgtg gttgcccggc tttcgcctcg 21780

ataatgtagg ttttatggcc ggttgtgagg atgaggtcgc cttcgtcttc gcggccgttg 21840

aggtggaggc gttcgatatt gtgtccggtg tcgcgtagct ggtggaggag tcttgtttcc 21900

cattcggctc cggcccgccg gttgcgtgcc tgctgtgtgg ccatagtttt ttagagtcct 21960

ttgtgtgttg tggtcatgtt ccagggctgt ttttcggcga gtggcccgaa gaatgtgtat 22020

tcggggtatg ctctgagtcg ttcgtatcgg gtgccgtcgg ggctggattt gcctgtgcgc 22080

tgtttgagta cggcgatgcg tgcctctgcc ggtatcgata gcccgttgcc gttatcctcg 22140

ccaccataca atgagactcc gaggatgagt tgtggttttt cggagaggcc gtttttgatt 22200

tctcgccgtg ctggcgggtg ttcgatgtcg gttccggttt tgtcggttgc gtggtgtgtg 22260

acaataatgg tggagccagt atccctgccc aatgctgtga tccattgcat ggcttcttgc 22320

tgtgcctggt agtcggattc gcagtcttga atgtccatca ggttgtcgat aacaatgagt 22380

ggtgggaagg tgttccacat ttccatgtag gcttgtaact ccatggtgat gtctgtccat 22440

gtgatgggtg actggaatga gaatgtgatg tgttggccgt ggtggatgct gtctcgatag 22500

tattctggcc cgtagtcgtc gatgttttgt tgtatttgtt gggtggtgtg ttgtgtgttg 22560

agggagatga ttcgtgtgga ggcctcccag ggtgtcatgt cccctgatat gtagagggcg 22620

ggctggttga gcatcgctgt gatgaacatg gctagccctg atttttggct gccggaccgc 22680

cccgcgatca tcaccaagtc gcccttatgg atgtgcaaat cttggttatc atatagtggt 22740

gcgagttgtg gtatgcgggg tagttcggct gcggtttggg aggctctctc gaaggatcgt 22800

tgtagagaga gcatcgggac cttaatctat ctgtctgttg gttgtgtggc tggtcagatg 22860

gagtcgatat cgatatcagc atcagcagag gctgaagtgt catctagctg accattatcg 22920

cgcttgtcta cgtattcggc aaccttatcg tagatggcgt cgtccaatgt tttgagcacg 22980

accgcgttga aaccgttttt ggtgcgcacg gtggctagtt tgaaggcctg ctcctcgcca 23040

aggtatgcct ctagttcgcg gatcatggag tgtgggcggt cgttattgcc gcgggctttc 23100

tcaataatag cgttggggat ggtttctggg gtgccgttgt tgagatcgtc tagggtgtgg 23160

aagatggtga catcagcgta gatgcggtct gcgacctgtc caccgtagcc ttcagtgttg 23220

tgctggacgt cgtgcacttt gaaggcgatg gccgtggcgt cctggtttcg ggaggggttg 23280

aagaaggtgc tgttgctgtt gttgcggtag tttgcgagtc ccataactat tgtttccttt 23340

tactgttgtg tctgtttttg ttggcttata ttggtttatc gggtgaggct gtttcgctta 23400

gtgcggaaag cgtcggaaac atcactgtta ctggtgatga tcttcttgta ctgttttaga 23460

aggtctgcta gctgtgcctt gcttgttgca ttgttgattt tgttgatgac gatggtgttt 23520

tctttggatg cgattttgtt gacgtagtct ttggctgcct ggttgtatcg gtcttggagg 23580

atgattgatg cgctcgctac gagtgttgct agatcccagt ctttggacac gtcatcgttt 23640

ttgagtccgc ctagcaggtc gatgatggcc tgttttgtct gctctgctgt gtctcctcgg 23700

atgaccgccc atggtgcagc atagtctcca ccatatttga gtgtgatcgt gagtcgatca 23760

ttgtcgatct tgtctttatc tgtcatttgg tgtccttttc tttattgtct gtttctggtg 23820

gctgtacggt ggattctacc gggtatctgt acgagttttt gccgttgacg gcccagcagg 23880

cgtctcgtac ggggcatcct ttacagagtg ttgtgacgtg ggggacgaag atgccttcgc 23940

tgattccttt cattgcttga ctgtacatgg atgatacatg ccggtaggtg ttgttgtcaa 24000

ggtcgtagag ttcggtggat gtgccttgtg tcggggactt gtcgtcgttg cggctggtgg 24060

ctggcgtcca aaacatgcct ttcgtgacat ggatgtcgtg ttggttgagc atgtaccggt 24120

atgtgtgcag ctgcatactg tcggcgggta ggcgtccggt tttgaggtcg aggatgaagg 24180

tttcgccggt gtcggtgtcg gtgaaaacac ggtcgatgta gccgactatt tttgtgtcat 24240

cgtcgaggat ggtttctacc gggtattcga tgcctggttt accgtccagg attgcggtga 24300

tgtattctgg gtggttgcgc ctccatgttt tccagcggtc cacaaaggtg gggccgtaaa 24360

ccatccacca gtcgtagtct ttcttgtgtg gtccgcctga ctcgcacatg tttttgcata 24420

ttctgccgga gggtttgatt tctgtgcctt cggattcggc gagggctacc tgggtgtcga 24480

aaatgttttt gaaggatgag agtttgtctg gcagtgcagg gtattcggcg ggattgtaca 24540

ggtgtaggtc gtattgttcg gtgatgtggt gtatggcgct tccggcgatg gtggcgtacc 24600

aggtgtggtg ttgggcgtga tagccgtggg ataggcgcca tttttctccg cattcggccc 24660

actgggtgag tgaactgtag gagatgtgtc ctgggtggct gatggttttc gggtattgtg 24720

ctagaggcat tacttgtcgc ttgtgttcca tgtgttgcgg gtgtcttggc cggcgtggtg 24780

ttgctggtag gcgaggagtg cgaggcagtg ccaggctgcg tgtgctagat ggggtagccc 24840

ggattcgtgg tcgaggttgt tgccttgctg ccatgatagt agatgcctgt agagggcgtc 24900

gacactgtgg ctccacgggt atcctccggt ccagttgttg tcgccatatt tggtggcacc 24960

gtatccggct acttcgccta gggcgtgaag ggatgctggg tcgatgaggg agagcctgca 25020

gagtttcaat tcttttcggg caccgctgtt ggggtcggtg tacatgcggg tgggctcatc 25080

catggggtgt gtgctcctta agggtgggtt actggttgtt gttgtgggct agggcggcgg 25140

cgagaataat gatggcgagg gtttcggcta tcagtatggg tgttgtgatc atttggtgtc 25200

tcggggattg ttggtgagtg ttgaggcacc caggagggtg gcgagggcgc atgcggcaat 25260

aatggcgagg gctgccttgt gtggggtgcc ggttgcgtac atccatgtga tgatggcacc 25320

ttggatccag gctaggctgg tgaagaaggt ttcgtagctg tgcagctcaa tgttgttgtt 25380

gggtgtgttc atgcttgctc ctgaagaatg gtgttgatgg ttttataaat gttgtacagg 25440

tcggtttcga tagataacag ttggttgatt tggtggtcga gatcaatgtc tgggttgagt 25500

gtgttgatgc gggaggcaat atcggtggct gtgcgtagtg tgccgccggt gtggtgaata 25560

atgtgtgccg tgtcggcgag tccggtggtg acggcgtagt gggataggag aggcatagcg 25620

gggatgctcc ttggcgggtt actgttgcgg gttgatgttg aggtcggtga cgtgcggtga 25680

gttttctgtt ccggtgacga ggcagtggac ggtgacgggt agtttggatg ctcccggctg 25740

gcggacggtg gcgccgtaga cgatgctgaa tgtgtcttta ccgatggttt tgtggagttg 25800

gaggtcgatg tcggggttgc cgttccagtt gacaccttgc gctgcggcct gttgttcggc 25860

tttgtggttg caggtgtgtg ctgccgtgat catggtgagt ccggtggcgg tttcttcacc 25920

ccttgcttgg gcttgcttgt gggctttggc ctgctcggct tgtagggatc gggtggcggc 25980

tgcctgccgt gccgctttct cggctttgcg ctgttgggta gtcttggggg tccatgtggt 26040

gttggctgtg gttgcctgtg gggctggctg tgaggtgagt ggcgggttgt cgtctggtgc 26100

tggcatgaat gaggcggcgg caatgatggc ggctgtgatg cctgcgatgg tgtagccgtt 26160

tttcttgttc atgttttgtg tcccctttcc ggggtgttgt tcgttgctga catggttaat 26220

actttcagcg gctgggccca ctgtcaaggc tgcgctcagt ttgtgtgagc gtttggtgtg 26280

tggctagggg ttttgtcatg taagcgtgac atgtcactac cttgcgtcca gtatccatgg 26340

cggttgcgag ccatcccttt ggcgagcatc tcgtccacag tgaggcacct gcggcgattg 26400

gggccttcct tgaccccgtg atcgcctatg cggtgcatgt ccccggcata agtgccatta 26460

aatgtttcgt ggcagactgt gcagtgttct ggtcggtatc cgatgattgt gctatcgcac 26520

ttgtggcatg tccattgcat gattggtcct tctttcgtgt tttaagcttg tgctctgagg 26580

attagagcga ctttcagccc ttgggggtag gattatatag gtcaggtatt tctaggcgat 26640

tctaggctca ttgtgtgtgg ttggggtttt atcgggcgca tagggttagc aggtggccca 26700

cattggtgcg gctcacattc cagtagagtt gcgtggcttc cttactggtg agcggcttcc 26760

actcgtcatg gctgaacacg gtgccatcgg atgcgatgaa cgtgttgggg cgtagcttgt 26820

gaagctcggc ttccacatgc tgccggtagg cttcggcgag gctctcaaaa tccatgtggt 26880

cgcaggagag gttttcgagg cgtgtcaggt cgaaaggctc cgggcagtcg tagctggctg 26940

gagtgtagag ctgggtgaag tggtcggcga tcttctgcat ggcgggttcc tttctggtgt 27000

gtggatggtt tttatcgtgt ggatgcgaca aggatggcgt ctacgtcgat catgtcgatc 27060

atgtcgttga gttcctcggc ctcattctcg gagaggtggc gccagtcggg tggcccgtat 27120

acggcgccgt cgagggtgac agtccacagg ggccggatga gtcgtatggc ttcttcgact 27180

ttggcgtggt acatgcggcg caccatatcc agatcgatgt cgtctgaatg gtttccggtg 27240

aggctgtgga ggctgagcgg gtcgatttct gtctgcctgt agaggctggt gaatgatggt 27300

gtgatgagtg tgccatccat gagtgtgctc ctttctaggg gttgttgtgg tttctagagt 27360

gtgtgggctg tgaccccaca gtcaaggcta cgctcatttg gattgagcgt ttcatatggg 27420

tgtggcatgg aatctacacc ctcatactgt gtgagatgta tcacatcccc ctggcttggt 27480

gtgcacccct caagactact ctgccgacct ggcgtggagg gtgtagccca gaaatgccgt 27540

ttaaagcttc aggggtacgc ctaggagcgc cttacagggt gggggctagg tatttatacc 27600

cccagcatat tctgatcgat tctagacgac tcccagagcc cgatacacga tcaaccatct 27660

cgacatagac catcagcccc tatcctggtt agctaagcct caactatgtg gacagtgtgg 27720

gacactgtgg gggaagaagg acacggtaca agaaagaggg gggagcatca gccttaaagc 27780

cttaagatct tagcgcttag caccgatggt cttagcagtt agcaccgagc ccttgagggg 27840

gctcggcatc agcctcatcg ggctcagctc atcaggcaca gccctgaaaa gggtacacgc 27900

catcagggaa ggcttgagag tacgaggagc cctagcgacg agtactcgaa agcctgaggg 27960

aacaccctca gtactgatga gcctagcgta ttcggaaagg acgcaagagt aaagtgtgac 28020

agctatccgg gagtgaaacc cgttccgact aggggtttca gccttaacca ccctcaaagg 28080

ttacaagact ctaagaaaat ttaagaaact tcttaggaag aaagttgtgt tcatatcccc 28140

ctaaaaacac ccaaaatagt cctcaaaccc gcctatagag ccaaacagtc aagtttgact 28200

cgtctagacg gcgtatgata ggctggacag gtagccagct ggacgcaagg ccagaaagtg 28260

ctgacgcact tcccgacctc gcttaccatc agtctaccaa acactttaaa gcttcaaggc 28320

ttagcgctaa gcccttaaga tcttaacgct tagcaccgag cccccctcaa gggctcgaca 28380

tcagtcttaa agtcttaaac actttaagta actttaaagc ttcaaggctt agcccttaag 28440

gatctaagtt actataaaag ctttaaacac ttaaagtaac tataaagctt taagagctta 28500

acatttaagg atataaataa acattaaagc tttaaagtct taaagtaaat atataacctt 28560

aacacttaag ttaagtataa aaccttaaag gcttagcact taaggatata aacttaacat 28620

cagtgtttaa gacttaaaga gttaaagtaa ctattaagac ttaaaggctt ataagcttta 28680

atactttaag tagctataag actttaaaaa cctgaagtac ttaaagttaa ccatcagtct 28740

taaactttaa tattataagt attaaagctt ataagttata aaagttttta gaagagttaa 28800

agggttaact tctttacttc tcttctctct ttggttcttt ctctcttctc ttcttttctt 28860

catcagggga gaagaggaac ctttaaccgt caacgctgat ggacttttca ccgtgtgact 28920

cgtgtgcttc tggtcgcaag ctcccatcgc acactcccca cactctttca cccgtgcccc 28980

tttacggctt agcgtgttcg tcggaaggcg tacggcgtgt cacgcttaaa cccttaacac 29040

caggtaagac ttaaagtgca tattataagt agaagacttt aaaacctata aggtgttccc 29100

gcttagcccg tgttccttta acgctaggcg ctcagcgcta agatgtgaaa cgtgaacacc 29160

catccacccc catttttctt ccgtgtcctt ctccttttga caccgctggg gggcgatgtg 29220

atatttctca catgccaggg ggtagtggag aaaacaacca ccccggaacg tttaagacac 29280

cccctcaaac gaacaaaaca gggcctagaa tcgatcagca gggcaccggt agggtattcc 29340

tacccccaga cgattcaagg ccattacagg agcaatgaga ggctcacagg ggccatggga 29400

gattgggggg cgtgatggca cacaccaacc gcacagccag ccaagcccac cggcgctggc 29460

gggcaaggct catcacccaa gcccgacaac aaggccaaac cgaatgccca ctctgcggag 29520

tcaccatcac ctggaacacc cacgacctgc caaccagccc cgaagccgac cacatcacac 29580

ccgtcagccg gggaggactc aacaccctcg acaacgggca aatcatctgc agaacatgca 29640

acagaagcaa aggcaacaga acacaaccaa acatcaaatt ccaacaacaa accacaaaaa 29700

cattgattcc atggtgaaaa acccgccaac ccccaccggg cacaccccct gcacacccgt 29760

gcaagacc 29768

<210> 24

<211> 11979

<212> DNA

<213> artificial sequence

<220>

<223> pANS514 plasmid

<400> 24

catggttgcg ccatgcagca caggccaagc gtgaggccca agcacgagga ctcgcccgct 60

gcccactgtg cggcgtctgg atggactacg aggtcggcaa gcgacccaac tcggccgaag 120

cagaccacat cagaccgcat tcgcttggtg gttcagacga catcgacaac attcgcgtca 180

tttgtcgtcg ttgcaatcaa tcgcgcggaa acggcctgaa gcgaccaggg cgccaacgtc 240

agcgtccaat caagcgcatc gagctggccc aaccggcccg cagtggggca tttcctgccc 300

cgccggcatg aatggaaggg cagtgcggat ggtgcggtcg ggcattcgat cgtgcccgga 360

cgggtcgccc gcgacgcttc tgctcggccc gctgtcgggt cgccgcgtcc cggtgtgcga 420

tcccgctggc catgaggtcc cgcactgcgt gggtccgctg cgacggcaag cgccccatca 480

ccctggctgg cgctccggcc tcatccacgg acccgggcac atggtctggc tggtcgcagg 540

tgcgacgcgc cacggccggc gatggcttcg ggaccatgct cggtgacggg ctggggtgct 600

gggatctcga ccacttcgac gatcagggcg cccgggcctt catcgaccgg atcgataagc 660

cgatcatctt cgccgagcgg tcggtgtcgg ggcatggctt ccacatcttc gtccggactg 720

acgaggcccc cggacgccgc accggaaaca tcgagttcta ctcacgccat cggttcatca 780

gggtcacagg agaccagttc gtctgaaggt cgtgccgggt ttgtgtctga tgaggagtgg 840

ttgtttctca tggatgctgc ggtgattcat gatgtggtgt ggcgtgaggg tcgcgcggat 900

ttggtggctt cgttgcgtgc tcatgtgaag gcttttatgg gtatgttgga taggtattcg 960

gttgatgtgg cgtctggtgg ccgtggtggg ggttctgcgg tagcgatgat tgaccggtat 1020

aggaagcgta ggggggcttg agtaggtgtc tggtgttgtt gggtctcagg ttcctcgtca 1080

ccgggtggct gtggcgtatt cggtgtctgc tggcggggat gctggggagc ttggtagggc 1140

ttatgggttg acgcctgatc cgtggcagca gcaggtgttg gatgattggc ttgctgtggg 1200

tggtaatggc aggcttgctt cgggtgtgtg tggggtgttt gttccgcggc agaatggcaa 1260

gaatgctatt ttggagattg tggagttgtt taaggcgact attcagggtc gccgtatttt 1320

gcatacggct cacgagttga agtcggctcg taaggcgttt atgcggttgc ggtcgttttt 1380

tgagaatgag cggcagtttc ctgacttgta tcgtatggtg aagtcgattc gtgcgacgaa 1440

tggccaggag gctattgtgt tgcatcatcc ggattgtgcc acgtttgaga agaagtgtgg 1500

ttgtccgggt tggggttcgg ttgagtttgt ggctcgtagc cggggttctg ctcgcgggtt 1560

tacggttgat gatttggtgt gtgatgaggc tcaggagttg tcggatgagc agttggaggc 1620

tttgcttcct accgtgagcg ctgccccgtc tggtgatcct cagcagattt ttttgggtac 1680

gccgccgggg ccgttggctg acgggtctgt ggtgttgcgt cttcgcgggc aggctttgtc 1740

gggtggtaaa cggtttgcgt ggacggagtt ttcgattcct gacgagtctg atccggatga 1800

tgtgtcgcgg cagtggcgga agttggcggg tgacactaat ccggcgttgg ggcgccgcct 1860

gaatttcggg acagtctcgg atgagcatga gtcgatgtct gctgccgggt ttgctcggga 1920

gcggcttggc tggtgggatc gtggccagtc tgcttcgtct gtgattccgg cggataagtg 1980

ggttcagtcg gctgtggttg aggcggctct ggttggcggg aaggtttttg gtgtctcgtt 2040

ttctcgctcg ggggatcgtg tcgcgttggc tggtgctggt aaaacggatt ctggtgtgca 2100

tgttgaggtt attgatggcc tgtctgggac gattgttgat ggtgtgggcc agctggctga 2160

ttggttggcg ttgcgttggg gtgacactga aaaggttatg gttgcagggt ctggtgcggt 2220

gttgttgcag aaggctttga cggatcgtgg tgttccgggt cgtggcgtga ttgtggctga 2280

tactggggtg tatgtggagg cgtgtcaagc cttcctggag ggtgtcaggt ctgggagcgt 2340

gtctcatcct cgtgccgatt cgaggcgtga catgttggat attgctgtga ggtcggctgt 2400

gcagaagaag aagggttctg cgtggggttg gggttcctcg tttaaggatg gttctgaggt 2460

tcctttggag gctgtgtctt tggcgtatct tggtgcgaag atggcgaaag cgaagcggcg 2520

tgaacggtct ggtaggaagc gggtgtctgt ggtatgaact cggatgagtt ggctctgatt 2580

gagggcatgt acgatcgtat tcaagggttg tcttcgtggc attgccgtat tgagggctac 2640

tatgagggct ctaatcgggt gcgtgatttg ggggttgcta ttccttcgga gttgcagcgg 2700

gtgcagacgg tggtgtcatg gcctgggatt gcggtggatg ctttggagga gcgtctggat 2760

tggcttggct ggactaatgg tgacggctac ggtttggatg gtgtgtatgc tgcgaatcgg 2820

cttgctacgg cgtcgtgtga tgttcacctt gatgcactga tttttgggtt gtcgtttgtg 2880

gcgatcattc cccaagagga tgggtcggtg ttggttcgtc ctcagtcgcc gaagaattgt 2940

actggccggt tttctgccga tgggtcttgt ttggatgctg gccttgtggt gcagcagacg 3000

tgtgatcctg aggttgttga ggcggagttg ttgcttcctg atgtgattgt tcaggtggag 3060

cggcggggtt cgcgtgagtg ggttgagacg ggccgtatcg agaatgtgtt gggtgcggtt 3120

ccgttggtgc ctgttgtgaa tcgtcgccgt acttctagga ttgatggccg ttcggagatt 3180

acgaggtcta ttagggctta cacggatgag gctgttcgca cactgttggg gcagtctgtg 3240

aatcgtgatt tttatgcgta tcctcagcgt tgggtgactg gcgtgagcgc ggatgagttt 3300

tcgcagccgg gttgggttct gtcgatggct tctgtgtggg ctgtggataa ggatgatgat 3360

ggtgacactc cgaatgtggg gtcgtttcct gtgaattctc ctacaccgta ttctgatcag 3420

atgcgtttgt tggcgcagtt gactgcgggt gaggcggctg ttccggaacg ctatttcggg 3480

tttatcactt ctaacccgcc ttctggggag gctttggctg cggaggagtc tcggcttgtg 3540

aagcgtgctg aacgcaggca gacgtcgttt ggtcagggct ggctgtcggt tggtttcctg 3600

gctgcccggg cgttggattc gagtgttgat gaggccgcgt tttttggtga tgttggtttg 3660

cgttggcgtg atgcgtcgac gccgactcgg gcggctacgg ctgatgctgt gacgaagctt 3720

gtgggtgctg gtattttgcc tgctgattct cggacggtgt tggagatgtt gggtttggat 3780

gatgtgcagg ttgaggctgt gatgcgtcat cgtgccgagt cttcggatcc gttggcggca 3840

ctggctgggg ctatttcccg tcaaactaac gaggtttgat aggcgatggc ttcgggtgct 3900

gtgtcgaggc ttgctgcgac tgagtatcag cgtgaggctg tcaggtttgc tgggaagtat 3960

gcgggctatt atgccgagtt gggtcgtttg tggcgtgccg gcaggatgag tgacacgcag 4020

tatgtgcgtt tgtgtgtgga gttggagcgt gccggccatg acggttcagc agctatggcg 4080

ggcaaattcg tttcagattt tcgccggttg aatggtgtcg atcctggttt gatcgtgtat 4140

gacgagtttg atgctgcggc ggctttggct aggtcgtttt cgactatgaa gattatgaat 4200

agtgacccgg atagggcgaa tgatacgatt gatgcgatgg ctgcgggtgt taatcgggct 4260

gttatgaatg ctggtcgtga cacggttgag tggtcggcgg gtgcgcaggg taggtcgtgg 4320

cgtcgggtga ctgatggtga tccgtgtgct ttttgtgcca tgttggctac gaggtcggat 4380

tatacgacta aagagcgggc gcttactact ggtcatacgc ggcgtcataa gcgtgccggt 4440

aggcgtccgt ttggttcgaa gtatcatgat cattgtggtt gtacggtggt tgaggttgtt 4500

ggtccttggg aaccgaatag ggctgatgcc gagtatcaga ggacgtatga gaaggctcgt 4560

gagtgggttg atgatcatgg gttgcagcag tcgtctggca atattttgaa ggctatgcgt 4620

actgttggtg gcatgagata atttgatgtg gtttccggtt gtgtgccgcc ggttatcggt 4680

gcacagggtt gtctcccgca cgggggtcaa caatgttgtg ttgttttccg caaggagtgt 4740

agggttaggc tatggccgat cagagtattg aggaacagaa tgttgacaat gatgttgtgg 4800

agtccggaaa ggataacggc attgttgata cagtaaaaga cgatggcggg caggaggtag 4860

ccgacaatca gttgaagaat gaaggcgagg gtaaatcgcc ggggactgat tggaaggcgg 4920

aggcccgtaa gtgggagtct cgtgctaaaa gtaatttcgc cgagttggag aagcttcgta 4980

catcgagtga cgattctgga tctactattg atgagcttcg ccgcaagaat gaggaactcg 5040

aagaccggat taacgggttt gttcttgagg gtgtgaagcg cgaggtggct gccgagtgtg 5100

gcctgtcggg tgatgcgatc gcttttcttc acggtagcga taaggagtcg cttgccgagt 5160

ctgctaaggc tttgaagggt ttgatcgacc atagtagtgg tggtggcgcg ggtgtgcgcc 5220

gtcttgcggg gagtgccccc gttgatgatg ttaaacgacg tgagggtgtc gcgtttgtgg 5280

atgctcttgt caataattct aggagatgat ttatcatggc tgacgatttt ctttctgcag 5340

ggaagcttga gcttcctggt tctatgattg gtgcggttcg tgaccgtgct atcgattctg 5400

gtgttcttgc taaactgtca ccggagcagc cgactatttt cgggcctgtt aagggcgccg 5460

tttttagtgg tgttccgcgc gctaagattg ttggcgaggg cgatgttaag ccttccgcta 5520

gcgttgatgt ttctgcgttt actgcgcagc ctatcaaggt tgtgactcag cagcgtgtct 5580

cggacgagtt tatgtgggct gacgccgatt accgtctggg tgtgcttcag gatctgattt 5640

ccccggccct gggtgcttct attggtcgcg ccgttgatct tattgctttc catggtattg 5700

atcctgctac gggtaagcct gctgcggctg tcaaggtgtc gctggataag acgaataaga 5760

cggttgatgc caccgattcc gctacggctg atcttgttaa ggctgttggt ctgattgctg 5820

gtgctggttt gcaggttcct aacggtgttg ctttggatcc ggcgttctcg tttgctctgt 5880

caactgaggt gtatccgaag ggttcgccgc ttgccggtca gccaatgtat cctgccgccg 5940

ggttcgccgg cctggataat tggcgcggcc taaatgttgg ttcttcttcg actgtttctg 6000

gtgccccgga gatgtcgcct gcttctggtg ttaaggctat tgttggtgat ttctctcgtg 6060

tccattgggg gttccagcgt aacttcccga ttgagctgat cgagtatggt gacccggatc 6120

agacggggcg tgacttgaag ggccataatg aggttatggt tcgtgccgag gctgtgctgt 6180

atgttgcgat tgagtcgctt gattcgtttg ctgtcgtgaa ggagaaggct gccccgaagc 6240

ctaatccgcc ggccggtaac tgattcattt gttgcgataa tgtttatgct gtgtgcaggg 6300

ggtggtgttg atgggtatca ttttgaagcc tgaggatatt gagcctttcg ccgatattcc 6360

tagagagaag cttgaggcga tgattgccga tgtggaggct gtggctgtca gtgtcgcccc 6420

ctgtatcgct aaaccggatt tcaaatatag ggatgccgct aaggctattc tgcgtagggc 6480

tttgttgcgc tggaatgata ctggcgtgtc gggtcaggtg cagtatgagt ctgcgggccc 6540

gtttgctcag actacacggt cgaatactcc tacgaatttg ttgtggcctt ctgagattgc 6600

cgcgttgaag aagttgtgtg agggtgatag tggggctggt aaggcgttca ctattacacc 6660

gaccatgagg agtagtgtga atcattctga ggtgtgttcc acggtgtggg gtgagggttg 6720

ctcgtgcggg tcgaatatta acggctatgc tggcccgttg tgggagatat gatatgaccg 6780

gttttcctta cggtgaaacg gttgtgatgc ttcagccgac tgttcgtgtc gatgatcttg 6840

gtgacaaggt ggaggattgg tctaagcctg tcgagactgt gtaccataac gtggccatct 6900

atgcttccgt ttcgcaggag gatgaggccg cggggcgtga ctcggattat gagcattgga 6960

cactgctgtt caagcagcct gtcaaggctg ctggttatcg gtgtcgttgg cgtattcggg 7020

gtgttgtgtg ggaggctgac gggtctccta tggtgtggca tcatccgatg tctggctggg 7080

atgctggtac gcaggttaat gtgaagcgta agaagggctg atgggttgtg gcacgtgatg 7140

ttgatgtgaa gctgaacttg ccgggtattc gtgaggtgtt gaagtcttct ggggtgcagg 7200

gcatgttggc tgagcgtggt gagcgtgtca agcgtgcggc ctcggcgaat gtgggcggta 7260

acgcttacga tagggcccag tatcgtgccg ggttgtcgtc tgaggtgcag gttcaccgtg 7320

ttgaggctgt ggcgcgtatt ggcaccacct ataagggtgg taaaaggatt gaggctaagc 7380

atggcacgtt ggcgaggtcg attggggctg cgtcgtgatc gtttacggtg atcctcgaat 7440

atgggctaaa cgtgtgttgg cggatgatgg ttggctgtct gatgtaccgt gcacgggtac 7500

tgtgccggat acatttgagg gtgatctgat ttggttggcg ttggatggtg gcccggagtt 7560

gcatgttcgt gagcgtgttt ttttgcgtgt gaatgtgttt tcggatacgc cggatcgtgc 7620

tatgtctttg gctcgccggg ttgaggctgt gctggctgat ggtgtggatg gtgatccggt 7680

ggtgttttgc aggcgttcga ctgggcctga tttgctggtg gatggtgcac gttttgatgt 7740

gtattcgctt tttgagctga tatgtaggcc tgcggagtct gaataagctt attgtttttg 7800

ttttaatgta attgtttgat atttaatggg ggttgtgatg gctgctacac gtaaagcgtc 7860

taatgttcgt tcagcggtta ctggcgacgt ttatattggt gacgcgcacg cgggtgattc 7920

tattaagggt gtggaggcgg ttccttccgg gcttacagct ttggggtatc tgtctgatga 7980

cgggtttaag attaagcctg agcgtaaaac ggatgatttg aaggcttggc agaatgcgga 8040

tgttgttcgc actgtggcta cggagtcgtc tatcgagatt tctttccagc tgattgagtc 8100

gaagaaggag gttatcgaac tgttttggca gtcgaaggtt actgccggat ctgattcggg 8160

ttcgttcgat atttctcctg gtgccacaac aggtgttcac gccctgttga tggatattgt 8220

tgatggcgat caggttattc gctactattt ccctgaggtt gagctcattg atcgtgacga 8280

gattaagggc aagaatggcg aagtgtacgg gtatggtgtg acgttgaagg cgtatcctgc 8340

ccagattaat aagactggta atgcggtgtc gggtcggggg tggatgacgg ctttaaaagc 8400

tgatactcct ccgactcctc cgccggcccc ggttcctccg aagcctcagc cggatccgaa 8460

tccgccgtcc ggtaactgat acacgatttt aggggattgt taatagatga gtgacactgg 8520

tttcacgttg aagattggtg atcgtagctg ggtgttggcg gatgcggagg agacggctca 8580

ggctgttcct gcccgcgttt tccgtcgtgc cgccaggatt gcccagtcgg gggagtctgc 8640

ggatttcgcc caggttgagg tgatgttttc tatgttggag gctgccgccc cagctgacgc 8700

ggtggaggcc ctggaggggc ttcctatggt tcgtgtggcg gaggttttcc gtgagtggat 8760

ggaatacaag cctgacggta agggtgcctc gctgggggaa tagtttggct ccacggcctg 8820

attgatgatt atcgtggggc catcgaatac gatttccgca ccaagtttgg tgtttctgtt 8880

tatagtgttg gtggcccgca gatgtgttgg ggtgaggctg tccggctggc tggcgtgttg 8940

tgtaccgata cgtctagcca gttggcggcc caccttaatg gttggcagcg cccgtttgag 9000

tggtgcgagt gggctgtgtt ggacatgttg gatcattaca ggtctgctaa tagtgagggg 9060

cagccggagc ctgtggcgag gccgactgat gagcgtcggg caaggtttac gtctgggcag 9120

gtggacgata ttttggcgcg tgttcgtgcc ggtggcgggg tgtctcgcga gattgatatt 9180

atggggtgaa tagtgtatgt ctggtgagat tgcttccgca tatgtgtcgt tgtatacgaa 9240

gatgcctggc cttaaaagtg atgttggtaa acagttgtcg ggtgttatgc ctgctgaggg 9300

gcagcgttcg ggtagcctgt ttgctaaagg catgaagttg gcgcttggtg gtgcggcgat 9360

gatgggtgcc atcaatgttg ctaagaaggg cctcaagtct atctatgatg tgactattgg 9420

tggcggtatt gctcgcgcta tggctattga tgaggctcag gctaaactga ctggtttggg 9480

tcacacgtct tctgatacgt cttcgattat gaattcggct attgaggctg tgactggtac 9540

gtcgtatgcg ttgggggatg cggcgtctac ggcggcggcg ttgtctgctt cgggtgtgaa 9600

gtctggcggt cagatgacgg atgtgttgaa gactgtcgcg gatgtgtctt atatttcggg 9660

taagtcgttt caggatacgg gcgctatttt tacgtctgtg atggctcgcg gtaagttgca 9720

gggcgatgac atgttgcagc ttacgatggc tggtgttcct gtgctgtctt tgcttgccag 9780

gcagacgggt aaaacctcgg ctgaggtttc gcagatggtg tcgaaggggc agattgattt 9840

tgccacgttt gcggctgcga tgaagcttgg catgggtggt gctgcgcagg cgtctggtaa 9900

gacgtttgag ggcgctatga agaatgttaa gggcgctttg ggctatttgg gtgctacggc 9960

tatggcgccg tttcttaacg gcctgcggca gatttttgtt gcgttgaatc cggttattaa 10020

gtctatcacg gattctgtga agccgatgtt tgctgccgtc gatgctggta tccagcggat 10080

gatgccgtct attttggcgt ggattaaccg tatgccggct atgatcacga gaatgaatgc 10140

acagatgcgc gccaaggtgg agcagttgaa gggcattttt gcgagaatgc atttgcctgt 10200

tcctaaagtg aatttgggtg ccatgtttgc tggcggcacc gcagtgtttg gtattgttgc 10260

tgcgggtgtg gggaagcttg ttgcagggtt tgctccgttg gcggttgcgt tgaagaatct 10320

gttgccgtcg tttggtgctt tgaggggtgc cgccgggggg cttggtggcg tgtttcgcgc 10380

cctgggtggc cctgtcggga ttgtgatcgg cttgtttgcg gcaatgtttg ccacgaacgc 10440

ccagttccgt gccgctgtta tgcagctggt ggctgtggtt ggtcaggcgt tgggccagat 10500

tatggcagct gtgcagccgc tgtttggttt ggttgctggc gtggttgcca ggttggcgcc 10560

ggtgttcggc cagattatcg gtatggttgc tggtttggct gcccggctgg tgcctgttat 10620

tggtatgctt attgcccggc tggttcctgt tatcacccag attattggta tggtaaccca 10680

ggttgctgcc atgttgttgc ctatgctgat gccggttatt caggctgttg ttgctgtgat 10740

acggcaggtt attggtgtca ttatgcagtt gatacctgtt ttgatgccgg ttgtgcagca 10800

gattttgggt gctgtcatgt ctgttttgcc gccgattgtt ggtttgatac ggtcgctgat 10860

accggtgatc atgtcgatta tgcgtgtggt ggtgcaggtt gttggtgctg tgctacaggt 10920

ggtggcccgt attattccgg ttgttatgcc gatttatgtt tcggtgattg gattcattgc 10980

caagatttat gctgcggtta tcgtttttga ggctaaggtt attggcgcta ttcttcgtac 11040

tattacgtgg attgtgaatc attcagtgtc tggcgtgagg tctatgggca cggccatcca 11100

gaatggctgg aatcatatta aatcgtttac gtctgcgttt attaacggtt ttaagtcgat 11160

catttctggc ggcgtgaacg cggttgtggg gttttttacg cggcttggtt tgtcggttgc 11220

ttcccatgtg aggtccggtt ttaacgctgc gaggggtgct gtttcttccg ccatgaatgc 11280

tattcggagt gttgtgtctt cggtggcgtc tgctgttggc gggtttttca gttcgatggc 11340

gtctcgtgtt cggaatggtg ctgtgcgcgg gtttaatggt gcccggagtg cggcttcttc 11400

tgctatgcat gctatggggt ccgctgtgtc tagtggtgtg catggtgtgc tgggtttttt 11460

ccggaatttg cctgacaata ttcggcgtgc gcttggtaat atggggtccc tgttggtgtc 11520

ggctggccgt gatgtggtgt ccggtttagg taatggtatc aagaatgctt tgagtggcct 11580

gttggatacg gtgcgtaata tgggttctca ggttgctaat gcggcgaagt cggtgttggg 11640

tattcattcc ccgtctcggg tgtttcgtga cgaggttggc cggcaggttg ttgccggttt 11700

ggctgagggt attactggta atgctggttt ggcgttggat gcgatgtcgg gtgtggctgg 11760

gaggctgcct gatgcggttg atgcccggtt tggtgtgcga tcgtctgtgg gttcgtttac 11820

cccgtatggc aggtatcagc gcatgaatga taagagtgtt gtggtgaatg tgaatgggcc 11880

tacttatggg gatcctgccg agtttgcgaa gcggattgag cggcagcagc gtgacgcttt 11940

gaacgcgttg gcttacgtgt gattttgggg gtgtggtgc 11979

<210> 25

<211> 83

<212> DNA

<213> artificial sequence

<220>

PAC7 cos for pAN594, < 223-

<400> 25

acaaaaggga ggtatttcac taagccgtac gaggtcttgc acgggtgtgc agggggtgtg 60

cccggtgggg gttggcgggt ttt 83

<210> 26

<211> 4670

<212> DNA

<213> artificial sequence

<220>

<223> operon of gp15-gp19+gp45

<400> 26

cgacgcggcg gtctgccgac ccggcaacga ccaactcccc gacgggcgct gacaccggcc 60

cggcagcgtg catgcgtgca tttccaccct caagaaccat tgactggcga cgcgcaggtg 120

ggagaattga actgaacgct ttgaacgcgt tggcttacgt gtgattttgg gggtgtggtg 180

catgtttatt cctgacccgt ctgatcgttc tggtttgact gtgacttggt ctatgttgcc 240

gttgattggt aatgatccgg agcgtgtgct tcatttgacg gattatacgg ggtcgtctcc 300

gataatgttg ttgaatgatt cgttgcgcgg tttgggtgtt cctgaggtgg agcatttttc 360

tcaaactcat gttggggtgc atggctcgga gtggcgcggg tttaatgtga agcctcgcga 420

ggtgacgcta ccggtgttgg tgtcgggtgt tggcccggat ccggtgggcg gttttcgtga 480

cggttttttg aaggcgtatg acgagttgtg gtctgctttt cctcctggcg aggtggggga 540

gttgtctgtg aagactcctg ccggtcgtga gcgtgtgttg aagtgccggt ttgattcggt 600

ggatgacacg tttacggtgg atccggtgaa caggggttat gcgcgttatc tgttgcattt 660

gacggcttat gacccgtttt ggtatgggga tgagcagaag tttcgtttca gtaacgctaa 720

gttgcaggat tggttgggtg gcggccctgt cgacggtaag ggtaccgcgt ttccggtggt 780

gttgacgcct ggtgttggtt cgggttggga taatctgtct aataagggtg atgtgcctgc 840

gtggcctgtg attcgtgttg aggggccgtt gtcgtcgtgg tctgtgcaga ttgatggttt 900

gcgtgtgtcc tcggattggc cggtggagga gtatgattgg atcactattg atacggatcc 960

tcgtaagcag tctgcgttgt tggacgggtt tgaggatgtg atggatcgtt tgaaggagtg 1020

ggagtttgcg cctatcccgc ctggcggttc tcggagtgtg aatattgaga tggttggttt 1080

gggtgccatt gttgtgtcgg tgcagtacag gtttttgagg gcttggtgaa tagttgatgg 1140

ctggttttgt tccgcatgta acattgttta caccggatta tcgccgtgtg gcgcctatca 1200

atttttttga gtcgttgaag ttgtcgttga agtggaatgg tttgtccact ttggagttgg 1260

tggtgtctgg tgatcattct aggcttgacg ggttgactag gccgggtgcg cggcttgtgg 1320

ttgattatgg tggtggccag attttttctg ggcctgtgcg tcgggtgcat ggtgtgggtc 1380

cgtggcgttc ttcgcgtgtg actatcacgt gtgaggatga tattcgtctg ttgtggcgta 1440

tgttgatgtg gcctgtgaat tatcgtcctg gtatggttgg tatggagtgg cgtgcggatc 1500

gggattatgc ccattattcg ggtgcggcgg agtcggtggc taagcgggtg ttgggggata 1560

atgcttggcg ttttccgtct ggtttgttta tgaacgatga tgagagtcgt ggccgctata 1620

ttaaggattt tcaggtgcgg tttcacgtgt ttgccgataa gttgttgccg gtgttgtcgt 1680

gggctcggat gactgtcacg gtgaaccagt ttgagaatgc gaagtttgat cagcgtggtt 1740

tggtgtttga ttgtgtgcct gctgtgaccc ggaaacatgt gttgactgcc gagtcgggtt 1800

cgattgtgtc gtgggagtat gtgcgtgacg ccccgaaggc gacatctgtg gtggttggtg 1860

gccgtggcga gggtaaggat cggctgtttt gtgaggatgt tgattcggcg gccgaggatg 1920

attggtttga tcgtgtcgag gtgtttaagg atgcccgtaa cacggattcc gagaaggtgt 1980

ctctcttcga tgaggctgag cgggtgttgt ccgagtcggg ggctacgtcg gggtttaaga 2040

ttgagttggc tgagtcggat gtgttgcggt ttggtcccgg caatctgatg cctggggatt 2100

tgatctatgt ggatgtgggt tctgggccta ttgcggagat tgtgcggcag attgatgtgg 2160

agtgtgtatc gcctggtgat ggttggacga aggtgactcc ggttgcgggg gattatgagg 2220

ataatccgtc ggccctgttg gctcgccgtg tggctggttt ggctgcgggt gtgcgggatt 2280

tgcaaaagtt ttagtaagtg attggggttt gttgtgggta ttgtgtgtaa agggtttgat 2340

ggtgtgttga ccgagtatga ttgggctcaa atgtctggtc tgatgggtaa tatgccgtct 2400

gtgaaggggc ctgacgattt tcgtgtcggc acgacgattc agggttctac ggtgttgtgt 2460

gagatcctgc cggggcaggc ttgggctcac ggggtgatgt gcacgtcgaa tagtgttgag 2520

acggtgacgg gtcagcttcc gggcccgggt gagactcgat acgactatgt ggtgttgtct 2580

cgggattggc aggagaatac ggccaagttg gagattgttc ccggtgggcg tgcggagcgt 2640

gccagggatg tgttgagggc tgagcctggc gtgtttcatc agcagctact ggcgactttg 2700

gtgttgtcgt ctaacgggtt gcagcagcag ttggataggc gtgctgtggc ggctagggtt 2760

gcgtttgggg agtctgctgc gtgtgatcct acccctgtgg agggtgaccg tgtgatggtt 2820

ccttcggggg ctgtgtgggc taaccatgcc ggcgagtgga tgttgttgtc tcccaggatt 2880

gagacgggtt cgaagtcgat catgtttggt ggttctgctg tgtatgctta cacgatcccg 2940

tttgagcgcc agttcagtag tccgcctgtt gtggtggcgt ctatggctac ggcggctggg 3000

ggcacggcac agattgatgt gaaagcctac aatgtgactg cccaaaattt tagtttggcg 3060

tttattacga atgatggttc gaagccgaat ggtgtgcctg cggtggcgaa ttggattgct 3120

gtcggcgtgt gactgcacgg gtgttgtggc ggatggtgtg atgttggggg gctgtggtgt 3180

cgtggtttac tcctgcactg gtggcctcta tttgtaccgc gttggccacg gttttgggtt 3240

ctgttcaggc tgtcacatcc cggtctagga agcgtttacg caggctgtcg gctcaggtgg 3300

atgcgatgga agagtatacg tggggtgtgc ggcgcgaggt gcgaaggttt aacgccgggc 3360

ttcctgatga tgtggagccg atgcatcttc ctgatttgcc cgagtttttg aaagatactg 3420

ttgatggtgg aggtgagtag ggttgaggga gttggaggag gagaagcggc agcgccgcaa 3480

ttttgagaag gcttcactgg tgttgttgtt tttgtcgctt gtgttgttgg cggtggttgc 3540

tgcgggtgct ttgcgtttcg gggctgtatc ctctgagcgg gattcggagc aggcgagggc 3600

ccagtcgaat ggtacggctg ccaggggttt ggctgcccgt gtgaagcagg cgtgtgcttc 3660

gggtggggtg gagtctgtgc gtcttcaccg ttctggtttg tgtgtggatg ctgtgcgtgt 3720

tgagcagcgt gttcagggtg tgccgggtcc tgccggtgag cgcggcccgc aaggcccttc 3780

aggtcctgcc ggccgggatg gtgttaatgg ttcggctggg ctggttggcc ctgttggtcc 3840

gcaaggttct ccgggtttga atggtgtgaa aggtcctgac ggcttgcctg gcgctaacgg 3900

ttcggatggc cgtgatggtg ttccaggtcg tgcaggtgct gacggtgtga acggcgttga 3960

cggcgctgat ggtcgggatg gttctgccgg tgagcgcggc ccgcaaggcc cttcaggtcc 4020

tgccggcccg caaggtgcac agggtgaacg gggtgagcgt ggtcccgccg gtgcgaatgg 4080

atcggatggc catgatggta aggatgggcg ctcggtggtg tctgtgtact gttccggggg 4140

ccgcctggtt gtgaaatata gtgacggtgt ggcttccacg atatcgggtt cggcggcctg 4200

ccagggtgtg aaaccgtcgc ctctagtgac tatatcatcc cacaaataga ggctcacagg 4260

ggccatggga gattgggggg cgtgatggca cacaccaacc gcacagccag ccaagcccac 4320

cggcgctggc gggcaaggct catcacccaa gcccgacaac aaggccaaac cgaatgccca 4380

ctctgcggag tcaccatcac ctggaacacc cacgacctgc caaccagccc cgaagccgac 4440

cacatcacac ccgtcagccg gggaggactc aacaccctcg acaacgggca aatcatctgc 4500

agaacatgca acagaagcaa aggcaacaga acacaaccaa acatcaaatt ccaacaacaa 4560

accacaaaaa cattgattcc atggtgagga tatccacgag ctgcgttcgg ctaaacccaa 4620

aagtaaaaac ccgccgaagc gggttttaac gtaaaacagg tgaaactgac 4670

<210> 27

<211> 1910

<212> DNA

<213> artificial sequence

<220>

<223> pAN241 vector

<400> 27

caagtggccc atcgaagagg acggcaccac catctcgccg ggcaagctca aggacgtgtc 60

caggctgacg ctcacggtgc tgctgcaccc ctcgtgcgcc atcatcgtgg atccccaaga 120

ttgtccggac ggcggttgag cgcggcctga taggcgccgc agctcctgct cccgggccgc 180

cccggtcggc ggtttactcc tttcctgccg gccggggcac tcaagacaac cgggggccct 240

cgcgaaattg aggggccccg cctgattgca agggggtgcc catgaagcaa cccgggcccc 300

accaaagaat gcgggctacc ttcaaggccg acaggggctg gcgagtggca tgcccacggt 360

gcgcctggca tgccaccagc acccaccttg catggctcat ggatcaggcc agcacacaca 420

cctgtgcacc cctgctgttg tcgcccacgc cacccgacgt ggagctggca ccggcaggcg 480

acgggctgtc cgtcctgtgg cccgaggtgg acggtgacgt gcagttcacc tgcatccaca 540

ccagcaccgc cacgtgcagg caggacgcac catgagcacc agtcgcaccg gcacggccac 600

atggttgcgc catgcagcac aggccaagcg tgaggcccaa gcacgaggac tcgcccgctg 660

cccactgtgc ggcgtctgga tggactacga ggtcggcaag cgacccaact cggccgaagc 720

agaccacatc agaccgcatt cgcttggtgg ttcagacgac atcgacaaca ttcgcgtcat 780

ttgtcgtcgt tgcaatcaat cgcgcggaaa cggcctgaag cgaccagggc gccaacgtca 840

gcgtccaatc aagcgcatcg agctggccca accggcccgc agtggggcat ttcctgcccc 900

gccggcatga atggaagggc agtgcggatg gtgcggtcgg gcattcgatc gtgcccggac 960

gggtcgcccg cgacgcttct gctcggcccg ctgtcgggtc gccgcgtccc ggtgtgcgat 1020

cccgctggcc atgaggtccc gcactgcgtg ggtccgctgc gacggcaagc gccccatcac 1080

cctggctggc gctccggcct catccacgga cccgggcaca tggtctggct ggtcgcaggt 1140

gcgacgcgcc acggccggcg atggcttcgg gaccatgctc ggtgacgggc tggggtgctg 1200

ggatctcgac cacttcgacg atcagggcgc ccgggccttc atcgaccgga tcgataagcc 1260

gatcatcttc gccgagcggt cggtgtcggg gcatggcttc cacatcttcg tccggactga 1320

cgaggccccc ggacgccgca ccggaaacat cgagttctac tcacgccatc ggttcatcag 1380

ggtcacagga gaccagttcg tctgaagaag ggggtgcgcc atggctgcac aggtcagggc 1440

cgtggacccc gatgagcgcc cacccgcccg caagcgggcc aagaccatca cccaggccgc 1500

gaagtccggc actgaggttg aactgttgga ggcactgcag gctcgcgtgg cccgcgccgt 1560

gcaggaccgt gacactccgc cgcgcgatct ggcagcgctg acgaagcggc tgatggacat 1620

cacccgggag ctcgaggcgg cccgggtcaa ggatcaggag gcgggatctg atggtgccgt 1680

caccgcagac gaaacatggc gaccgcaagc tctctgaggt cgccaagcac ctgatccttc 1740

ctgaagggat cgtctcgacg ggctggccgg ccgtgcgtga ccggtgtggc gagtggggtg 1800

tggtcttcga ccgttggcag gacggcatgg gccgggtgat cctgtcgaag cgcggcagcg 1860

gcctgttcgc cgctggtgtg ggcggggtcg gcatgtcgat cccgcgccag 1910

Claims (28)

1. A producer bacterial cell for producing lytic phage particles or a lytic phage-derived delivery vehicle, said producer bacterial cell stably comprising at least one phage structural gene and at least one phage DNA packaging gene, said phage structural gene and phage DNA packaging gene being derived from a lytic bacteriophage,

wherein the expression of at least one of said phage structural genes and/or at least one of said phage DNA packaging genes in said producer bacterial cell is controlled by an induction mechanism.

2. A producer bacterial cell according to claim 1, wherein the bacterial cell further comprises a payload to be packaged into the phage particle or phage-derived delivery vehicle.

3. A producer bacterial cell according to claim 2, wherein the payload is a nucleic acid payload comprising a packaging site derived from the lytic bacteriophage.

4. A producer bacterial cell according to any one of claims 2 to 3, wherein the payload is to be delivered into the targeted bacterial cell.

5. The producer bacterial cell of claim 4, wherein the payload is stably maintained in the targeted bacterial cell.

6. The producer bacterial cell of claim 4, wherein the payload is not replicated in the targeted bacterial cell.

7. A producer bacterial cell according to any one of claims 4 to 6, wherein the payload comprises a sequence of interest.

8. The producer bacterial cell according to claim 7, wherein said sequence of interest only plays a role in said targeted bacterial cell.

9. The producer bacterial cell according to claim 8, wherein said target bacterial cell is from a species or strain different from said producer bacterial cell.

10. A producer bacterial cell according to any one of claims 1 to 9, wherein the same induction mechanism controls the expression of at least one of said phage structural genes and at least one of said phage DNA packaging genes.

11. A producer bacterial cell according to any one of claims 1 to 9, wherein the expression of at least one of said phage structural genes and the expression of at least one of said phage DNA packaging genes are controlled by different induction mechanisms.

12. A producer bacterial cell according to any one of claims 1 to 11, wherein said at least one induction mechanism controls the expression of all of said phage structural genes.

13. A producer bacterial cell according to any one of claims 1 to 12, wherein said at least one induction mechanism controls the expression of all of said phage DNA packaging genes.

14. A producer bacterial cell according to any of claims 1 to 13, wherein said induction mechanism further controls the copy number of said at least one of said phage structural genes and/or said at least one of said phage DNA packaging genes.

15. The producer bacterial cell of any of claims 2-14, wherein said at least one induction mechanism further controls the copy number of said payload in said producer bacterial cell.

16. The producer bacterial cell of any of claims 2-14, wherein another induction mechanism controls the copy number of said payload in said producer bacterial cell.

17. A producer bacterial cell according to any of claims 1 to 16, wherein said phage structural gene and phage DNA packaging gene derived from said lytic bacterial phage are comprised in at least one plasmid, chromosome and/or helper phage.

18. The producer bacterial cell according to any one of claims 1 to 17, further comprising at least one gene involved in phage regulation derived from said lytic bacteriophage.

19. A producer bacterial cell according to any of claims 1 to 18, wherein said producer bacterial cell further comprises at least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation.

20. A producer bacterial cell according to claim 7, wherein said producer bacterial cell is from the same bacterial species or strain as the bacterial species or strain from which said non-lytic bacteriophage is derived and/or to which said non-lytic bacteriophage is targeted.

21. The producer bacterial cell according to any one of claims 1 to 20, wherein said producer bacterial cell is an e.

22. The producer bacterial cell according to any one of claims 1 to 20, wherein said producer bacterial cell is a propionibacterium freudenreichii bacterial cell.

23. A producer bacterial cell according to claim 22, wherein said phage structural gene and phage DNA packaging gene are derived from a dermatophyte acne bacteriophage.

24. A method for producing lytic phage particles or a lytic phage-derived delivery vehicle, comprising:

(a) Providing a producer bacterial cell according to any one of claims 1 to 23, and

(b) Inducing expression of said at least one of said phage structural genes and said at least one of said phage DNA packaging genes in said producer bacterial cell, and assembly of products expressed by said at least one phage structural gene and said at least one phage DNA packaging gene, thereby producing lytic phage particles or a lytic phage-derived delivery vehicle.

25. A hybrid helper phage system comprising:

(i) At least one phage DNA packaging gene derived from a lytic bacteriophage,

(i') at least one phage structural gene derived from said lytic bacteriophage,

(i ") optionally, at least one phage gene involved in phage regulation derived from said lytic bacteriophage, and

(ii) At least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation,

Wherein the genes (i), (i') and (ii) are contained in separate nucleic acid molecules or separate nucleic acid molecules, and

wherein the hybrid helper phage system does not comprise any expressed phage structural genes derived from the non-lytic bacteriophage.

26. The hybrid helper phage system according to claim 25, wherein the genes (i), (i'), (i ") and (ii) are contained in bacterial chromosomes.

27. The hybrid helper phage system according to claim 25, wherein the genes (i), (i'), (i ") and (ii) are contained in separate plasmids.

28. The hybrid helper phage system according to claim 25, wherein the hybrid helper phage system consists of a hybrid helper phage system comprising:

(i) At least one phage DNA packaging gene derived from a lytic bacteriophage, at least one phage structural gene and optionally at least one phage gene involved in phage regulation, and

(ii) At least one gene derived from a non-lytic bacteriophage involved in phage excision/insertion, phage DNA replication and/or phage regulation,

wherein the hybrid helper phage does not comprise any phage structural genes derived from the non-lytic bacteriophage.