KR102314100B1 - Novel Canine Adenovirus Type 2 Antigen and Uses Thereof - Google Patents

Abstract

The present invention relates to a novel canine adenovirus type 2 antigen and uses thereof, and more specifically, the present invention relates to a novel canine adenovirus type 2 strain deposited with accession number KCTC13956BP, An antigen for domestic isolated canine adenovirus type 2 enzymatic immunization technique and a method for detecting an antibody against canine adenovirus type 2 using the same are provided. According to the present invention, since the antibody titer to canine adenovirus type 2 can be quickly confirmed without a pretreatment process for a large amount of dog serum, the risk of canine adenovirus type 2 infection in dogs can be greatly reduced.

Description

Novel Canine Adenovirus Type 2 Antigen and Uses Thereof

The present invention relates to novel canine adenovirus type 2 antigens and uses thereof.

Mammalian-derived adenoviruses cause diseases of the respiratory, gastrointestinal and eye mucosa in many species of mammals, including humans. It is known that there are more than 50 adenoviruses in humans, 27 in monkeys, 2 in dogs and cattle, 4 in pigs, and 10 serotypes in cattle. Canine adenovirus exists in two serotypes, and belongs to the genus Mastadenovirus of the adenovirus family (Adenoviridae). Canine adenovirus type 1 causes infectious hepatitis in dogs, and adenovirus type 2 causes infectious laryngotracheitis. Canine adenovirus is easily transmitted from dogs not vaccinated with canine adenovirus vaccine, and because it is a virus that causes oral-fecal circulation, it is transmitted through direct contact with rice bowls, shelters, respiratory rhinorrhea, feces, and urine.

Canine adenovirus type 1 infection was first reported in wild otters and foxes who showed long-term anorexia and weight loss in Korea, and canine adenovirus type 2 infection was reported not only in domestic dogs but also in other countries. Recently, canine adenovirus type 2 infection, which is a form of respiratory disease, is prevalent both at home and abroad.

In addition, canine adenovirus type 2 does not have an envelope, and the size is 70-90 nm. According to whole genome analysis, canine adenovirus type 2 has a DNA genome of about 31 kb that encodes an initial protein, a structural protein, and a terminal protein, and has about 30 open reading frames (ORFs). Canine adenovirus type 2 particles are icosahedral, and are formed of a capsid composed of hexon, penton, and fiber. There are 252 capsids, and each capsid consists of 240 hexons and 12 pentons.

At the ends of the genome, there are repeats known as inverted terminal repeats (ITRs), which represent unique characteristics of viruses. Canine adenovirus type 2 binds to host cell receptors through viral fibers and enters the cell through endocytosis. The envelope is removed and the nucleic acid migrates into the nucleus where proliferation occurs. Transcription of viral DNA by cell-derived polymerase sequentially forms early and late mRNAs and encodes early and late proteins, respectively. About 12 nonstructural proteins are formed before the gene genome is amplified. Virus structural proteins are synthesized late, and DNA replication proceeds. Virus replication starts at both ends of the DNA. As the viral particles assemble in the nucleus, the cell's synthetic machinery automatically stops. Cellular DNA synthesis is stopped, cellular RNA and protein synthesis is stopped, and the cell dies. Because the polymerase of the host cell is required for DNA replication, the virus tends to proliferate well when the cell divides. The initially expressed proteins are E1, E2, E3, and E4, and the protein of E3 is known to be associated with pathogenicity. Fiber protein is composed of the domains of tail, shaft, and knob, and serves as a link between the receptor of the host cell and the adenovirus capsid.

Methods for testing canine adenovirus antibodies known so far include a virus neutralization test (VN), a hemagglutination inhibition test (HI), and an enzyme linked immunosorbent assay: ELISA. The virus neutralization test is accurate, but it is a method used in the laboratory and requires equipment and skilled personnel related to cell culture. It is cumbersome because it requires 25% kaolin and goose blood cell treatment to remove it. In addition, canine adenovirus type 1 has the property of aggregating blood cells of geese, chickens, and guinea pigs, so it can be used to perform a hemagglutination inhibition test. is difficult to manufacture. Currently, ELISA kits are not produced or licensed in Korea. Therefore, there is a problem in that it must be imported and used when performing serological investigation of canine adenovirus type 2 infection with an ELISA kit.

For the detection of antibodies to canine adenovirus type 2, it is ideal to detect antibodies using a neutralization test method, but this neutralization test method can be performed in a laboratory using cells and having a cell culture facility.

Therefore, in order to detect canine adenovirus type 2 antibody in a laboratory that does not have a cell culture facility, a rapid, convenient, and highly specific and sensitive antibody detection technique regardless of location and equipment is required.

Under these circumstances, the present inventors made intensive research efforts to develop a method for detecting blood antibodies to canine adenovirus type 2 quickly and conveniently regardless of location and equipment while having excellent specificity and sensitivity. As a result, a novel canine adenovirus type 2 was established by identifying canine adenovirus type 2 isolated in Korea and culturing it for 40 generations, and it was propagated and purified and used as an antigen against canine adenovirus type 2 by enzymatic immunization technique. The present invention was completed by identifying that blood antibodies to canine adenovirus type 2 can be rapidly and accurately detected in dogs when applied to

Accordingly, an object of the present invention is to provide a novel canine adenovirus type 2 strain deposited with accession number KCTC13956BP.

Another object of the present invention is to provide a method for preparing an antigen for enzymatic immunization.

In addition, another object of the present invention is to provide an antigen for the enzyme immunization technique prepared by the above method.

Another object of the present invention is to provide an enzyme immunization technique kit for detecting an antibody against canine adenovirus type 2 containing an antigen.

Another object of the present invention is to provide a method for detecting an antibody against canine adenovirus type 2 .

Other objects and advantages of the present invention will become more apparent from the following detailed description and claims.

The terms used herein are used for the purpose of description only, and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly dictates otherwise. In the present specification, terms such as “comprise” or “have” are intended to designate that a feature, number, step, operation, component, part, or combination thereof described in the specification exists, but one or more other features It should be understood that this does not preclude the existence or addition of numbers, steps, operations, components, parts, or combinations thereof.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiment belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present application. does not

Hereinafter, the present invention will be described in detail.

According to one aspect of the present invention, the present invention provides a novel canine adenovirus type 2 (Canine Adenovirus Type 2) strain deposited with accession number KCTC13956BP.

The Canine Adenovirus Type 2 strain entrusted with the accession number KCTC13956BP is a strain consisting of the nucleotide sequence of SEQ ID NO: 2; E1b 19K protein consists of the amino acid sequence of SEQ ID NO: 5; E3 12.5K protein is characterized in that it consists of the amino acid sequence of SEQ ID NO: 7.

In one embodiment of the present invention, the strain is a novel canine adenovirus type 2 strain isolated from the domestic canine adenovirus type 2 parent strain and subcultured for 40 generations, named APQA1701-40P, and Biotechnology Research Institute Biological Resource Center ( KCTC) on September 18, 2019, and was given an accession number KCTC13956BP.

In the present specification, the novel strain of the present invention, "Canine Adenovirus Type 2 strain deposited with accession number KCTC13956BP" is used interchangeably with APQA1701-40P.

In the present invention, "canine adenovirus type 2 (canine adenovirus type 2)" belongs to the adenoviridae and includes a virus that can infect canines and cause hepatitis or pneumonia, a respiratory disease, but preferably the APQA1701-40P strain. include

The APQA1701-40P is a virus strain obtained by isolating the virus from the canine adenovirus infection diagnosis sample provided by the Agriculture, Forestry and Livestock Quarantine Headquarters, culturing the virus under various conditions, and passing and propagating in the 40s. In comparison, there were changes in the nucleic acid sequence and the amino acid sequence, which resulted in adaptation to the environment and mycological properties with increased proliferation rate.

Further, according to another aspect of the present invention, the present invention provides a method for preparing an antigen for enzymatic immunization, comprising the steps of:

(a) inoculating the cultured cells with the accession number KCTC13956BP and canine adenovirus type 2 (APQA1701-40P) and culturing; and

(b) purifying the cultured canine adenovirus type 2 (APQA1701-40P).

1 shows the cytopathic effect of isolates of canine adenovirus type 2, and FIG. 2 is a diagram illustrating the fixation of the cells of FIG. 1 and confirming them with a fluorescent conjugate by reacting with a specific antibody of canine adenovirus type 2, FIG. 3 is a diagram comparing a part of the amino acid sequence of canine adenovirus type 2 with a virus that was passaged in their 40s and a virus before passage, and FIG. Each step of the method of the present invention will be described in detail.

Step (a) of the present invention is a step of inoculating and proliferating canine adenovirus type 2 in cultured cells, which are animal cells, and then culturing.

In the present invention, the cultured cells may include any cells known in the art as long as they can be used for infection or propagation of canine adenovirus type 2 .

Preferably, MDCK (Madin-Darby canine kidney) cells, primary dog kidney cells, or animal cells that are a combination thereof, but is not limited thereto.

In one embodiment of the present invention, the cultured cells are MDCK cells, and the MDCK cells have high proliferation efficiency of canine adenovirus type 2 and are easy to handle.

In one embodiment of the present invention, the MDCK cells proliferated 95% in the incubator are inoculated with APQA1701-40P strain, a culture medium that does not contain fetal bovine serum is added, and after 3-4 days of culturing, the cytopathic effect , and repeating the freeze-thaw process three times to release the intracellular virus, centrifuged at 3,000 rpm for 15 minutes, and harvested, followed by a purification step.

After incubation, before performing the purification step, centrifugation may be performed to remove dead or dying cells residues.

Step (b) of the present invention is a step of filtering the cultured canine adenovirus type 2 using a filter and then purifying using a column.

The filter can filter a size of 0.45 or 0.2 μm. In an embodiment of the present invention, canine adenovirus type 2 is filtered using a pump equipped with a 0.2 μm filtration filter.

For the purification, the filtered canine adenovirus type 2 is mixed with a binding solution (25 mM histidine, pH 6.0) at 1:4 and applied to the column. In the embodiment of the present invention, the ratio of the binding solution is 1:4, but it can be used in a ratio of 1:1 to 1:5.

After applying all of the binding solution to the column, 10 ml of the binding solution is applied to the column in order to wash the material not attached to the column. To recover canine adenovirus type 2 attached to the column, an elution solution (75 mM tris Cl, 525 mM Nacl, pH 8.5) is applied to recover the purified antigen by fraction. As shown in FIG. 5 , the antigen with the highest concentration was selected by measuring the protein concentration at 280 nm. The purified antigen thus recovered can be used as an antigen for enzymatic immunization. In order to confirm whether the purified antigen is canine adenovirus type 2, it was observed with an electron microscope. As shown in FIG. 6 , virus particles of canine adenovirus type 2 can be clearly seen. In order to obtain a more purified antigen, the above filtration purification step may be repeated.

In addition, according to another aspect of the present invention, the present invention provides an antigen for the enzymatic immunization technique prepared by the method for producing the antigen for the enzymatic immunization technique.

The antigen of the present invention is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 2; It has the amino acid sequence of SEQ ID NO: 3.

The antigen of the present invention may include not only the sequence of SEQ ID NO: 3 described herein, but also biological equivalents thereof. For example, additional changes may be made to the amino acid sequence of an antigen to further improve its biological properties. Such modifications include, for example, deletions, insertions and/or substitutions of amino acid sequence residues of the antigen. Such amino acid variations are made based on the relative similarity of amino acid side chain substituents, such as hydrophobicity, hydrophilicity, charge, size, and the like. By analysis of the size, shape and type of amino acid side chain substituents, arginine, lysine and histidine are all positively charged residues; alanine, glycine and serine have similar sizes; It can be seen that phenylalanine, tryptophan and tyrosine have similar shapes.

Therefore, based on these considerations, arginine, lysine and histidine; alanine, glycine and serine; And phenylalanine, tryptophan and tyrosine can be said to be biologically functional equivalents.

In introducing the mutation, the hydropathic index of the amino acid may be considered. Each amino acid is assigned a hydrophobicity index according to its hydrophobicity and charge: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cysteine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).

The hydrophobic amino acid index is very important in conferring an interactive biological function of a protein. It is a known fact that amino acids having similar hydrophobicity indexes must be substituted to retain similar biological activities. When introducing a mutation with reference to the hydrophobicity index, the substitution is made between amino acids exhibiting a difference in the hydrophobicity index, preferably within ±2, more preferably within ±1, and even more preferably within ±0.5.

On the other hand, it is also well known that substitution between amino acids having similar hydrophilicity values results in proteins having equivalent biological activity. As disclosed in US Pat. No. 4,554,101, the following hydrophilicity values are assigned to each amino acid residue: arginine (+3.0); lysine (+3.0); asphaltate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5±1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); Tryptophan (-3.4).

Amino acid exchanges in proteins that do not entirely alter the activity of the molecule are known in the art (H. Neurath, R.L. Hill, The Proteins, Academic Press, New York, 1979). The most commonly occurring exchanges are amino acid residues Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Thr/Phe, Ala/ exchange between Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu and Asp/Gly.

In consideration of the above-described variation having biological equivalent activity, it is construed that the nucleic acid molecule encoding the antigen of the present invention includes a sequence showing substantial identity to the sequence listed in the sequence listing. The substantial identity is at least 61% when the above-described sequence of the present invention and any other sequences are aligned as much as possible, and the aligned sequence is analyzed using an algorithm commonly used in the art. means a sequence that exhibits homology, more preferably 70% homology, even more preferably 80% homology, and most preferably 90% homology. Alignment methods for sequence comparison are known in the art. Various methods and algorithms for alignment are described in Smith and Waterman, Adv. Appl. Math. (1981) 2:482; Needleman and Wunsch, J. Mol. Bio. (1970) 48:443; Pearson and Lipman, Methods in Mol. Biol. (1988) 24: 307-31; Higgins and Sharp, Gene (1988) 73:237-44; Higgins and Sharp, CABIOS (1989) 5:151-3; Corpet et al., Nuc. Acids Res. (1988) 16:10881-90; Huang et al., Comp. Appl. BioSci. (1992) 8:155-65 and Pearson et al., Meth. Mol. Biol. (1994) 24:307-31. The NCBI Basic Local Alignment Search Tool (BLAST) (Altschul et al., J. Mol. Biol. (1990) 215:403-10) is accessible from NBCI et al. It can be used in conjunction with a sequence analysis program. The BLSAT can be accessed at www.ncbi.nlm.nih.gov/BLAST/. A method for comparing sequence homology using this program can be found at www.ncbi.nlm.nih.gov/BLAST/blast_help.html.

The present invention provides a nucleic acid molecule encoding the antigen, and includes the nucleotide sequence of SEQ ID NO:2.

As used herein, the term "nucleic acid molecule" has a meaning comprehensively including DNA (gDNA and cDNA) and RNA molecules. also includes modified analogs (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews, (1990) 90:543-584). The sequence of the nucleic acid molecule of the present invention may be modified. Such modifications include additions, deletions, or non-conservative or conservative substitutions of nucleotides.

The nucleic acid molecule of the present invention is construed to include a nucleotide sequence exhibiting substantial identity to the above-described nucleotide sequence. The substantial identity is at least 80% when the nucleotide sequence of the present invention and any other sequences are aligned to correspond to the maximum and the aligned sequence is analyzed using an algorithm commonly used in the art. means a nucleotide sequence exhibiting homology, more preferably at least 90% homology, and most preferably at least 95% homology.

Since the antigen for enzymatic immunization according to the present invention is highly purified through a purification step, a high correlation (r) value can be obtained when the enzymatic immunization technique is performed.

Since the antigen of the present invention is prepared by the above-described method, redundant descriptions are omitted to avoid excessive complexity of the present specification.

In addition, according to another aspect of the present invention, the present invention provides an enzyme immunization technique kit for detecting an antibody against canine adenovirus type 2 comprising the antigen.

By detecting the presence or absence of an antibody against canine adenovirus type 2 present in a sample using the kit, it is possible to detect whether or not a canine adenovirus type 2 infection and a specific antibody for canine adenovirus type 2 are present, and furthermore, it is easy to detect It can discriminate domestic strains quickly, simply, and with high specificity and sensitivity, confirming whether or not canine adenovirus type 2 infection and antibody production are present, and thus can be efficiently used for early diagnosis of canine adenovirus type 2 .

Further, according to another aspect of the present invention, the present invention provides a method comprising the steps of: (a) contacting a biological sample with a canine adenovirus type 2 antigen of the present invention, which is an antigen for enzymatic immunization; and (b) detecting the presence of the antibody bound to the canine adenovirus type 2 antigen in the sample by an enzyme immunization technique.

In the present invention, the term enzyme-linked immunosorvent assay (ELISA) includes those based on colorimetric, chemiluminescent and fluorescence analysis. Enzyme-linked immunosorvent assay (ELISA) includes small amounts of drugs and other antigenic components in samples including plasma and urine. It has been successfully applied during measurement and has the advantage of being simple to perform.

The "biological sample" is defined as any animal classified as a mammal, including any animal, such as humans, livestock and farm animals, and zoological, sporting or pet animals such as dogs, horses, cats, sheep, pigs, cattle, and the like. refers to a body sample isolated from an animal, preferably a dog, of In an embodiment, the biological sample is a sample from an animal infected with canine adenovirus type 2 or an animal vaccinated.

The sample may be a biological fluid such as serum, plasma, vitreous fluid, lymph fluid, synovial fluid, vesicle fluid, semen, amniotic fluid, milk, whole blood, urine, brain-spinal fluid, saliva, sputum, tears, sweat, mucus, tumor lysate , and tissue culture media, as well as tissue extracts such as homogenized tissue, tumor tissue, and cell extracts. Preferably, the sample is serum isolated from an animal. In the present invention, the serum is preferably a supernatant obtained by removing clots from blood obtained from animals, but excluding hemolyzed serum.

For detection of antibodies to canine adenovirus type 2, a biological sample is contacted with an antigen for the enzymatic immunization technique of the present invention. The antigen may be in a form coated (immobilized) on a support.

The antigen is preferably coated on the support at 10 μg/mL, but is not limited thereto. Coating (immobilization) is typically accomplished by pre-activation of the support with nitric acid and reducing agents known in the art as described in non-covalent or covalent coupling or by adsorption to a water-insoluble matrix or surface prior to the analytical procedure or This is accomplished either by insolubilizing the capture reagents, without activation, using glutaraldehyde or carbodiimide crosslinking, or by insolubilizing the capture reagents after the assay procedure, eg, by immunoprecipitation.

The solid phase used for immobilization may be any inert support or carrier that is essentially water-insoluble and useful in immunoassay assays, including, for example, supports in the form of surfaces, particles, porous matrices, and the like. Examples of commonly used supports include assay plates or test tubes made from polyethylene, polypropylene, polystyrene, etc., including small sheets, Sephadex, polyvinyl chloride, plastic beads, and 96-well plates, as well as, for example, filter paper , agarose, cross-linked dextran and other particulate materials such as polysaccharides.

In an example of the present invention, the 96-well plate was used as a solid phase and the antigen was coated at 4° C. for 12 hours.

In addition, for example, in order to coat an antigen on the plate of the enzyme immunization technique and stop the rest, 0.05% PBST (Na₂HPO₄ (1.15 g), NaCl (8 g), KCl (0.2 g), KH₂PO₄ (0.2 g), pH 7.2) ), and 0.05% Tween 20 and 5% skim milk in PBS can be prepared and used as a stop reagent (stop buffer), or 5% bovine serum albumin can be used.

To wash the plate of the enzyme immunization technique, preferably, add Na₂HPO₄ (1.15 g), NaCl (8 g), KCl (0.2 g), KH₂PO₄ (0.2 g), Tween 20 (0.5 mL) to make 1,000 mL and use the washing solution. It can be used, but is not limited thereto.

Dog serum can be used as the primary antibody in the enzyme immunization technique of the present invention, and in a specific embodiment of the present invention, it is confirmed that the optimal dilution factor of the serum used in the enzyme immunization technique is 20 to 200 times, preferably 100 times. did.

In the enzyme immunization technique of the present invention, peroxidase-conjugated anti-dog IgG can be used as a secondary antibody (conjugate), and when using it, the optimal amount used is 10 ng to 30 ng, preferably 20 ng. It was confirmed by example. Thereafter, when the OD (Optical Density) value is measured and analyzed according to the enzymatic immunization technique known in the art, the presence of an antibody bound to the canine adenovirus type 2 antigen is detected in a sample isolated from an animal simply and quickly. There is an effect that can be done.

Since the method of the present invention uses the antigens described above, redundant descriptions are omitted to avoid excessive complexity of the present specification.

The present invention relates to an antigen for enzymatic immunization technique of domestic isolated canine adenovirus type 2 obtained by purifying a novel canine adenovirus type 2 strain entrusted with accession number KCTC13956BP and canine adenovirus type 2 using the same. A method for detecting an antibody is provided. According to the present invention, since the antibody titer to canine adenovirus type 2 can be quickly confirmed without a pretreatment process for a large amount of dog serum, the risk of canine adenovirus type 2 infection in dogs can be greatly reduced.

1 is a diagram showing the cytopathic effect shown when the canine adenovirus type 2 used in the present invention is infected with cells.
2 is a diagram showing canine adenovirus type 2 used in the present invention infecting cells, reacting with a specific antibody of canine adenovirus type 2, and then staining with a fluorescent conjugate.
Figure 3a is a diagram showing the deletion of 6 amino acids in the E1b 19K protein of the APQA1701-40P strain by comparing the E1b 19K amino acid sequence of the canine adenovirus type 2 APQA1701 strain and the APQA1701-40P strain.
Figure 3b is a diagram showing a comparison of the E3 12.5K amino acid sequence of the canine adenovirus type 2 APQA1701 strain and the APQA1701-40P strain, wherein the 21st amino acid of the E3 12.5K protein of the APQA1701 strain is substituted from alanine to glycine.
4 is a schematic diagram illustrating a process of purifying canine adenovirus type 2 using a column.
5 is a diagram showing the protein concentration of a solution obtained by purifying canine adenovirus type 2 using a column.
6 shows canine adenovirus type 2 particles confirmed by electron microscopy in a solution obtained by purifying canine adenovirus type 2 using a column.
7 is a view showing the results of confirming the protein of a solution obtained by purifying canine adenovirus type 2 using a column by SDS-PAGE and confirming the protein by western blotting using a canine adenovirus type 2 specific antibody.
8 is a diagram showing the results of an experiment for determining the optimal concentration of canine adenovirus type 2 antigen in the enzyme immunization technique.
9 is a diagram showing the results of an experiment for determining the optimal dilution factor of the serum sample in the enzyme immunization technique.
10 is a diagram showing the correlation between the results of the virus neutralization test and the results of the enzyme immunization technique according to the present invention.
11 is a diagram showing an example of an enzyme immunization technique performed to detect canine adenovirus type 2 antibody.

Hereinafter, the examples are only for explaining the present invention in more detail, and those of ordinary skill in the art to which the present invention pertains that the scope of the present invention is not limited by these examples according to the gist of the present invention It will be self-evident for

Example 1. Obtaining a novel canine adenovirus type 2 strain

1-1. Canine adenovirus type 2 passage and propagation

The present inventors performed the following experiments to obtain a novel canine adenovirus type 2 strain.

First, the APQA1701 strain was isolated from dog tissues using MDCK cells. The selected APQA1701 strain was passaged by adding urea or DMSO to the growth medium as shown in Table 1 below. Urea was added to contain 5 mM in the D-MEM medium for proliferation from the 6th to the 20s, and proliferation and culture were performed.

Urea is a strong surfactant and has the property of breaking non-covalent bonds of proteins, which can increase permeability when viruses bind to receptors. However, as shown in Table 2 below, urea of 7.8 mM or more was toxic to MDCK cells, and the canine adenovirus type 2 was passaged from 6 to 40 years by adding it to the medium at a concentration of 5 mM.

DMSO (dimethyl sulfoxide) has the property of increasing cell permeability, so when added to the medium, DMSO was added to the D-MEM medium at 0.1% so that the virus could easily enter the MDCK cells. As shown in Table 1 below, DMSO was toxic to cells at 1% or more. However, it did not show cytotoxicity at concentrations below 0.5%. Therefore, the passage of canine adenovirus type 2 from 21s to 40s was performed by adding DMSO to 0.1%. As a result of confirming the virus content in the 40s passage ^{in the cells, 10 7.0} _{TCID 50} /mL was shown, which was used in the following examples.

number of passages passage cell Processing details note 1-5 MDCK cells - 10 ^4.5 TCID ₅₀ /ml 6-20 MDCK cells 5 mM urea 21-40 MDCK cells 5 mM urea, 0.1% DMSO

matter cytotoxicity Concentration (mM) 1000 500 250 125 62.5 31.2 15.6 7.8 3.9 Urea + + + + + + + + - Concentration (%) 10 5 2 One 0.5 0.2 0.1 0.05 0.01 DMSO + + + + - - - - -

1-2. Sequencing of canine adenovirus type 2, APQA1701-40P

The present inventors named the canine adenovirus type 2 passaged 40 times from the MDCK cells isolated in Example 1-1 as APQA1701-40P and deposited at the Institute of Biotechnology and Biotechnology Biological Resources Center (KCTC) on September 18, 2019. , was given accession number KCTC13956BP.

As a result of analyzing the nucleotide sequence of the entire gene of APQA1701-40P by NGS (Next generation sequencing) technique (Macrogen Korea), the number of open reading frames (ORF) of the gene sequence of APQA1701-40P was about 30, and a total of 31,053 It was confirmed that it consists of nucleotides, and the entire gene base sequence of the APQA1701-40P is shown in SEQ ID NO: 2, and its amino acid sequence is shown in SEQ ID NO: 3. In addition, the entire gene base sequence of the APQA1701 is shown in SEQ ID NO: 1.

As a result of confirming the homology of the base sequence and amino acids with the APQA1701 strain, it showed more than 99% homology, but 18 nucleotides were deleted in the sequence of the APQA1701-40P strain, and also deleted when compared with the deduced amino acid sequence It was confirmed that 6 amino acids of the modified site were deleted.

Deletion of these genes shortens the coding for the protein and can affect the growth and structure of the virus.

Specifically, it can be confirmed that the canine adenovirus type 2 APQA1701-40P strain passed in 40s has a deletion of 18 nucleotides encoding 6 proteins in the E1b 19K gene. The E1b 19K amino acid sequence of the APQA1701 strain encoded by the E1b 19K gene is shown in SEQ ID NO: 4, and the E1b 19K amino acid sequence of the APQA1701-40P strain is shown in SEQ ID NO: 5.

As a result of aligning the sequences, as shown in FIG. 3a, tryptophan, which is the 122nd amino acid, serine, the 123rd amino acid, arginine, the 124th amino acid, methionine, the 126th amino acid, of the E1b 19K amino acid sequence of the APQA1701 strain. It was confirmed that glutamine and amino acid 127, isoleucine, were deleted.

That is, the novel APQA1701-40P strain of the present invention is characterized in that amino acids 124 to 129 of the E1b 19K amino acid sequence represented by SEQ ID NO: 4 of the APQA1701 strain are deleted.

In addition, as shown in Figure 3b, in the novel APQA1701-40P strain of the present invention, in addition to the deletion site, the 21st amino acid of the E3 12.5K gene of the APQA1701 strain represented by SEQ ID NO: 6 is converted from alanine to glycine It was confirmed that the substituted variant (variant). The E3 12.5K amino acid sequence of the APQA1701 strain encoded by the E3 12.5K gene is shown in SEQ ID NO: 6, and the E3 12.5K amino acid sequence of the APQA1701-40P strain is shown in SEQ ID NO: 7.

Therefore, the strain APQA1701-40P, the strain of the present invention, which is canine adenovirus type 2 passaged in the 40s, is a novel strain with different changes in base sequence and amino acid compared to the original isolate, whereby the growth rate is fast and the growth environment It exhibits mycological characteristics that can be adapted to

Example 2. Preparation of antigen for using canine adenovirus type 2 for antibody test in enzyme immunization technique

2-1. Proliferation of canine adenovirus type 2

A high content of canine adenovirus type 2 is not only required to obtain the antigen of the enzymatic immunization technique, but the serum essential for the proliferation process interferes with the purification of the virus. Therefore, the MDCK cells are cultured in a rotary incubator until the cells proliferate 90%, the canine adenovirus type 2 (APQA1701-40P) is inoculated into the cultured MDCK cells, and 100 ml of serum-free medium is added to the roller bottle. After 4-5 days, the virus was centrifuged by repeating the freezing and thawing process 3 times. As a result of confirming the virus content of the canine adenovirus type 2 harvested in this way, 10 ^7.0 TCID ₅₀ /mL was shown. 100 mL of canine adenovirus type 2 prepared in this way was obtained.

2-2. Purification of canine adenovirus type 2

The use of purified antigens in enzymatic immunization techniques is key. Accordingly, the present inventors performed as shown in FIG. 4 to purify canine adenovirus type 2 for use as an antigen.

Specifically, the column used for purification is Nuvia cPrime (Bio-Rad). The attachment solution and washing solution used for this column were 25 mM histidine (pH 6.0), and the elution solution was 75 mM tris-Hcl, 52 5 mM NaCl (pH 8.5). Before applying the sample to the column, the proliferated antigen was filtered with a 0.2 μm or 0.45 μm filter to remove foreign substances. In a 250 mL sterilized plastic container, 40 mL of the propagated canine adenovirus type 2 was prepared by mixing 160 mL of the attachment solution, and 20 mL of the washing solution was flowed through the column. The above canine adenovirus type 2 and adhesion solution mixture was applied to the column. After flowing the antigen, the column was washed with 20 mL of the washing solution, the elution solution was applied, and the eluted samples were collected by 1 mL. As shown in FIG. 2 , the collected sample was measured for absorbance of the protein at 280 nm, and the protein was confirmed by SDS-PAGE. As a result of measuring the protein concentration of the elution solution, it was confirmed that it was 1.5 mg/mL in the fifth elution solution, and virus particles were confirmed with an electron microscope as shown in FIG. 6 . This was used as an antigen for indirect enzyme immunization.

2-3. Determination of the optimal concentration for use of canine adenovirus type 2 antigen

In order to evaluate the antigens identified in Example 2-2, the present inventors prepared serum from 165 dogs raised in Korea. In order to use the serum as a standard serum in the enzyme immunization technique, a serum neutralization test was performed. Through the result of the serum neutralization test, more than two-fold was confirmed as positive.

As described above, 8 of the sera confirmed in the serum neutralization test were selected and the optimal concentration of canine adenovirus type 2 antigen to be used in the enzyme immunization technique was set using the sera. Antigens were diluted to 50, 25, 12.5, 6.25, 3.12, 1.56, 0.78, 0.39, 0.19, 0.09, 0.04 μg/mL, dispensed in a 96-well plate at 100 μl, and coated at 4° C. for 12 hours. Prepare a coated 96-well plate with 200 µl of PBST [Na ₂ HPO₄ (1.15 g), NaCl (8 g), KCl (0.2 g), KH₂PO₄ (0.2 g), Tween 20 (0.5 mL) to make 1,000 mL) ], and using a blocking buffer containing 5% skim milk [prepared by adding 0.05% Tween 20 and 5% skim milk to 0.05% PBST solution], the antigen-uncoated area was cleaned It was stopped and incubated at room temperature for 2 hours. With dog serum, 100 μl of 5 positive serum (serum neutralizing antibody, 8, 32, 128, 256, 512) and 3 negative serum (2 fold or less serum neutralizing antibody) were diluted 100 times and applied. After 1 hour, washing was performed twice with PBST, and 100 μl of peroxidase-conjugated goat anti-dog IgG, a secondary antibody (conjugate), was diluted to 20 ng and applied. After reacting at 37°C for 1 hour, 100 μl of a coloring agent (2'2 zaino di 3-ethylbenzthiazoline: ABTS) was added, and after 10 minutes, color development was stopped with 1% sodium dodecyl sulfate solution. 8 shows.

As shown in FIG. 8 , the sera negative for canine adenovirus type 2 exhibited an absorbance value of less than 0.5. Through this, the optimal concentration for use in the enzymatic immunization technique against canine adenovirus type 2 in dog serum was set at 10 μg/mL.

2-4. Determination of the optimal dilution concentration of serum for canine adenovirus type 2 antibody test

In order to determine the serum dilution factor for canine adenovirus type 2 antibody test, the present inventors coated a 96-well plate with canine adenovirus type 2 antigen at a concentration of 10 μg/mL based on the results obtained in 2-3 above. and 5 positive and 3 negative serums with serum neutralizing antibody titers of 8, 32, 128, 256, 512 times to canine adenovirus type 2 (no more than 2 times the serum neutralization test antibody) were diluted from 20 times to 40,960 times. to 100 μl each, and reacted at 37° C. for 1 hour. After 1 hour of reaction, the mixture was washed twice with PBST, and peroxidase-conjugated goat anti-dog IgG, a color developer, and a color stop solution were performed in the same manner as in the experimental method of 2-3, and the results are shown in FIG. 9 .

As shown in FIG. 9 , it was confirmed that the canine adenovirus type 2 negative serum exhibited an absorbance value of less than 0.4, whereas the canine adenovirus type 2 positive serum exhibited an absorbance of 1.0 or more at a serological dilution factor of 8 or more.

Therefore, the optimal serum dilution concentration for testing canine adenovirus type 2 antibody in dog serum was set to 100-fold.

Example 3. Confirmation of Sensitivity, Specificity and Accuracy of Enzyme Immunization Technique for Canine Adenovirus Type 2 Serology Test

The present inventors confirmed whether an antibody against canine adenovirus type 2 could be detected in dog serum by the enzyme immunization technique performed under the optimal conditions obtained in Example 2 above.

The comparative test method for the enzyme immunization technique for serology of canine adenovirus type 2 is the serum neutralization test. For this serum neutralization test, a serum neutralization test was performed using the canine adenovirus type 2 propagated in Example 2-1.

For the serum neutralization test, 50 μl of serum was added to the first well of a 96-well plate, followed by binary dilution with 50 μl of medium, and 50 μl of 200 TCID ₅₀ /0.1 mL of virus was added to neutralize at 37°C for 1 hour. After that, 0.1 ml of 40,000 MDCK cells per 1 ml were added to each well, and serum neutralization was determined after 3-4 days. Specifically, 10 μg/mL of the canine adenovirus type 2 antigen purified in Example 2 was dispensed in a 96-well plate by 100 μl, coated at 4° C. for 12 hours, and washed with 165 dog serum 100 times. After dilution, 100 μl was applied. After that, it was carried out in the same manner as above.

The results obtained using the enzyme immunization technique and the results obtained in the virus neutralization test are shown in Table 3 below.

Based on the results in Table 3 above, the sensitivity, specificity, and accuracy were calculated as follows:

Specificity = 141/141*100 = 98.6%

Sensitivity = 19/19+3*100 = 86.4%

Accuracy = 141+19/165*100=97.0%.

As in the above calculation, the specificity, sensitivity and accuracy of the virus neutralization test were 98.6%, 86.4%, and 97.0%, respectively.

In addition, the results of confirming the correlation based on the results obtained in the virus serum neutralization test method and the absorbance results of the enzyme immunization technique are shown in FIG. 10 .

As shown in FIG. 10 , the R2 value was 0.8253 in the virus neutralization test method, and when it was converted back to the correlation (r) value, it was confirmed to be 0.91, confirming a high correlation.

As shown in FIG. 11 , when a serological audit was performed for canine adenovirus type 2 based on the above information, it was confirmed that an antibody positive reaction against adenovirus type 2 was actually performed.

Therefore, when the enzymatic immunization technique is performed using the canine adenovirus type 2 purified according to the present invention as an antigen, the antibody titer to the canine adenovirus type 2 can be checked simply by diluting the serum from a large amount of dog serum. .

Overall, the strain of the present invention was confirmed to be a novel canine adenovirus type 2 strain in which a deletion of a specific amino acid exists due to a sequence difference from the parent strain by continuously culturing the isolate isolated from the known strain for 40 passages. , In addition, when purified and used as an antigen, since the canine adenovirus type 2 antibody is accurately and easily detected, it is useful for greatly reducing the risk of canine adenovirus type 2 infection.

In addition, the sucrose method and the cesium chloride method, which are virus purification methods known in the art, chemically affect the virus by using a chemical reagent, and space and equipment that cannot be purified without an ultracentrifuge Although there are certain limitations, the present invention provides a method for rapid purification in a general laboratory that can use a small pump and column without these limitations.

In addition, the present invention relates to the welfare of companion animals and enables serological investigation of canine adenovirus type 2 among respiratory diseases in dogs. It is expected that a large amount of samples can be tested in a general laboratory in time, so that it has high industrial applicability.

Although the present invention has been described as the preferred embodiment mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also intended that the appended claims cover such modifications and variations as fall within the scope of the present invention.

Biotechnology Research Institute Biological Resource Center KCTC13956BP 20190918

<110> REPUBLIC OF KOREA (Animal and Plant Quarantine Agency) <120> Novel Canine Adenovirus Type 2 Antigen and Uses Thereof <130> QIA1-100P <160> 7 <170> KoPatentIn 3.0 <210> 1 <211> 31071 <212> DNA <213> Artificial Sequence <220> <223> APQA1701 nucleotide seq. <400> 1 catcatcaat aatatacagg acaaagaggt gtggcttaaa tttgggcgtt gcaaggggcg 60 gggtcatggg acggtcaggt tcaggtcacg ccctggtcag ggtgtttcca cgggaatgtc 120 cagtgacgtc aaaggcgtgg ttttacgaca gggcgagttc cgcggacttt tggctggcgc 180 cccgggtttc tgggcgttta ttgattttgc ggtttaccgg gtggtgcttt taccactgtt 240 tgcggaagat ttagttgttt atggagctgg tttcggtgcc agtttcccca cagctaatgt 300 gaaagtttat gtcaatataa cagaaacact ctgttctcag tttacagcac cccacccggt 360 ggttttttgc cacgcctttg ggttaatttt atttccctat acgcagcctt aaattctcag 420 tgcagacgaa agaggactac tcttgagtgc gcagcgagaa gagttttctc ttcactctgt 480 ctgatatatt ttctgaaaaa tgaaatatac tattgtgccg gcgccgcgca acctccatga 540 ttatgtttta gagctactgg aagagtggca gccggactgc cttgactgtg agtatcctca 600 tggcggcccc tcgccgccta ctctgcacga tctttttgat gttgagctgg agacttctca 660 cagccctctt gtgggcctgt gtgattcttg tgcggaggct gacactgatt cgagtgcgag 720 cactgaggct gattctggct ttagtccttt atccactccg ccggtttcac ctcttccacc 780 gcatcccacc tctcctgcta gcatttctga cgacatgttg ctgtgcttag aggaaatgcc 840 cacctttgat gacgaggacg aggttcgaag tgcggcgacc acctttgagc ggtgggaaaa 900 cacttttgac ccccatgtgg gtcctatttt tggctgtttg cgctgtgctt tttatcaaga 960 gcaggatgag aatgcacttt gtgggctttg ctatctaaag gcccttgctg aaggtaagtt 1020 ttaagttaat taaatgtttg ggcaggttaa atgttttagg tgtgtattga ttttaaattt 1080 tgctttttag tgccttttgc tttgcctgta cgttcagagc ccgcttcggc tggagctgag 1140 gaggaagatg atgaagttat ttttgtgtct gccaaacctg ggggcagaaa gaggtcagcg 1200 gctactccct gtgagccaga tggggtcagc aaacgccctt gcgtgccaga gcctgagcaa 1260 acagaacctt tggatttgtc tttgaagcca cgcccgcact aatctgtttg agtacaaagc 1320 aataaagtaa tcttgtttaa caagtttgcc tacatttgtg gttttacggg gcggggcgag 1380 tagtatataa tgccaaaagc cagtgcctgc ttcattaagc ttctagactg agctaagagc 1440 aggtagtatg gaccctctca agatttgtga aaactacctt acttttagag ctataattag 1500 gggaagtact ttgtcgcctg gattttttag gcggtggtgt tttcctgcct tggctgatgt 1560 ggtgggcaat atagtggaac aggaggaagg caggttttgg caaattttac ctgaaaacca 1620 cactttttgg ggtcttttgc gcaggggctt tactgttgct tcttttactg aaattattac 1680 agcagctcag ttggaaaata gaggtagaca gttggccttt ttagctttta tatcattttt 1740 gctacgcaac tggccttctg actctgtagt gcctgaagct gacagacttg acctggtctg 1800 tgcgccggca tggagcagaa tgcagatatg gagccagacc gccaggttaa tcaacgacct 1860 ccaagattcc gtgctcgagg agcaggggtc cgcggaagag gaagagtgcg aagaagcgct 1920 tttagcaggg gacagcgacg acccattatt cgggtagatg acttgcagct gcccgacccc 1980 ctgtatgtta tgcaagcttt gcaacgggac cacactttag aaatgcccag agggcaggta 2040 gattttagct ggattgaggc tgaagagagg cgggtaggtc ctacagacga gtggtacttt 2100 gaggctgtga agacttacaa agctaagccg ggagatgact tgcaaactat aatcaaaaac 2160 tatgccaaga tttccttaga atgtggggcc gtgtatgaaa ttaattctaa gattagggtt 2220 acgggggctt gctacattat tggtaattgt gctgtgctta ggcctaacct gcctgctgga 2280 gaagcaatgt ttgaggtttt gaatgttgat tttattccct ctattggttt tatggaaagg 2340 atagtgtttt ccaatgttat ttttgattgc aggaccaccg cgactgtagt gtgttgcatt 2400 agtgaaagaa acaccttgtt tcacaattgt gttttttctg gccctcacat gttatgtttg 2460 gaccttaggg cgggggcgga ggtgaggggc tgtcactttg tgggggcggt gtgtgcgttg 2520 cgtagcaagg ggctgtacag tattcgagtc aaaaatagca tttttgaaaa gtgtgctttt 2580 ggggtggtga ccgggtcaaa ggcttttatt agccattgca tgtttaagga ttgtacctgc 2640 tctattatgc tggggggtca gggcactatt gcccatagtc agtttattgt aactacttct 2700 gctgaggccc ccatgaacct gcaactgtgc acttgcgagg gtaatggaag tcatgtagtt 2760 ccattgggga atattcactt tgcttctcac cgggaagctt cgtggcctac gttttatgca 2820 aacaccttgg ttcgggtgcg cttgtatatg ggccggcgcc ggggagtttt tcaccccaag 2880 cagtctactt tgtcaatgtg tgtaattgca gcccctcggg gggttgtgca gagaatttat 2940 ttgtttggtg tgtatgatgc tacttgtgcc attatgcaac tgggcgaggc aggcaatgct 3000 gctagtgaaa gactgtgtac ttgcgggttc agacacagca ccccttccct gcgggccacc 3060 tatgtaactg acaccaggat tgaccgggag ctgaactctc aagacacggc tgagttcttt 3120 agcagtgatg aagataattt ttaggtgagt agatgggcgt ggtttggggg agtataaaag 3180 gggcgcggta cgtggctgtg tatttacagc catggaccct caacagaagg ggcttgtgaa 3240 cacgtgtttt gtgactacgc gtattccgtc ttgggcagga gcaagacaga atgtcaccgg 3300 gtcagattta gaaggaaagc ccgtgccctc agatgtgctg gaaagtggac gcccgcttgc 3360 agccccgcgc atcagaactt tgtatgagga gcagcagctg aacatgcttg cagtgaatgt 3420 tcttttggat gagctgaaga tccaggtggc tgccatgcaa aactctgtga ctgctattca 3480 gcgagaagta aatgatctaa agcaacgaat cgcccgagat taatgtaaaa ataaaattta 3540 tttctttttt gaatgataat accgtgtcca gcgttgtctg tctgtaatag ttctatgaat 3600 tctttccatg gcttggtaca ctttagcttg aatgttaagg tacatgggga tgagtccctc 3660 gtcggggctc aagtacagcc agtgcataca attgtcaatg ggctcagtat tgtatacaat 3720 ccaatcaaag ttagtattga ggcggtggta gttaaaaata tccttaagca ggagggaaat 3780 agcagtgggt tgtcccttag tgtagatgtt aataaaccta ttaagctggg aaggatgcac 3840 cttaggactg atgatatgca tctttgcttg gatttttaag ttggcaatgt ttcctccctg 3900 gtctcgtctg ggattcatgt tgtgcaagac cactataaca gagtagcctg tgcacttggg 3960 aaacctgtct agcagtttag agggaaaggc gtgaaagaac ttggctatgc ccctattgcc 4020 ccctaagtct tccatgcact catccattac aatagctatt ggccctttag aagcggccct 4080 cgcaaaaaca ttggtagggt tggtaacatc atagttggcc tcattggtga gttctgcata 4140 agacatttgt acaaacttgg gcatgatgct gccactctga ggaaccaccg tgttttcggg 4200 gcctgcaaga aagtttccct cacatatttg ggtttcccat gctgcaattt cctggggagg 4260 aatcatgtcc acctgggggg tgataaaaaa gactgtttct ggggcgggaa ctataagctg 4320 ggcagacatg aggttcctaa gaagctggga cttgccacac cctgtgggcc catacactgc 4380 agcaataaag ggttgcttgt agtagttaag ggatgcacag cttccatctt tattaaggta 4440 cttattgacc ttcatgcaac actgggttat gctatcttga atatctagta ggtcatttaa 4500 cagactgtct ccccccatag aaagcagttc ctcaaaagag gcaaagtttt tgaggggctt 4560 tagtccctct gcatatggca tgtttgttag gctttgctgg agagtttgca acctgtccca 4620 gaggcttttt aagtcatcca ctgcagctcc atccaggtag tttctgtgtt tcttgggttt 4680 ggatgcttgc ggctgtatgg aatgagacga tgcatgtcta gagcatgcag ggttttgtcc 4740 ttccatggcc tcagcttcct tgttagggtg gtttctgtca ctgtgaaagg ttgcacgtgg 4800 gcctgcacgc tggtgagggt cctgcgcagg ctcatcctgc tggttttgaa gacgtccgag 4860 ccctgctgca tgtcggcata gtagcaagct tttagcacat cataagagag ggtggaagtg 4920 gcatgacctt tggccctgag cttgcccttg cccacgtgat ggcacttagg gcaggtggtg 4980 tccttgagag catacaactt gggtgccagg tacactgatt ctgagctgta agcatccgat 5040 ccacacttct cacactgtgt ttcacactct actagccaag tgagctctgg gttgctggga 5100 tcaaagacta gcttgcctcc attttttttg agtcgtttct tacctttttc ttccatcaat 5160 ttgtaacctt ccatggtggt gaagaggctg tcagtgtctc cgtagacaga tttcaggacc 5220 ctgttttcaa gtggggttcc cgcgtcgttt tcgtagagaa actgagacca ctctgagaca 5280 aatgcccggg tccatgctag gacgaaggaa gcaatttgtg agggatatct gttattggct 5340 attaggggg tggacttttc taaagtatgt aaacaaaagt catcatcctc agcatcaata 5400 aacatgattg gcttataatg gtatgacgtc atgctaaaat tatgcataaa aggcgcttcg 5460 gcgagggggt ctttagttgg atcttcttcg ctgcaagaag gcggtgccaa ctggtgaacg 5520 tcagaattga caggtgggta cgctacgaca aattctggca tgatttctgc acacaaattg 5580 tctgtttcta tgtatgagga tgatttgact acataggcac cagaagcaat ttctttagcg 5640 atgtttgact ctatttgatc tgaaaaaact gtttttttgt tgtctagctt ggtagcaaaa 5700 gacccataga gggcatttga cagcagctta gcaatgctgc gcatggtttg atttttgctt 5760 ttgtctgctt cctctttggc actaatgttg agctgcacat actctctggc aagacatttc 5820 cactctggaa acacggtggt tctggtatca gggatgaggg taaccttcca cccgcggttg 5880 tgcagggtaa taacatctat agaggtcgca atctccccgc gcaggggctc gttggtccag 5940 caaagcctgc ctcccttgcg tgaacaaaat gggggcagca cgtccagaaa ctcttctgcc 6000 ggtgggtcag catcaattgt gaagatgccc ggaagaaggt ccttatcaaa gtagctaata 6060 gttttgtgat ttagcatttt ttgctcataa gcctttatgg ccagcgcccg ctcatagggg 6120 tttagcggtg aaccggcggg aaaagggtgt gtaagggcac ttgcatacat gccacatatg 6180 tcatagacaa acactggctc ttgcaacact ccaatgtaag tggggtagca cctgcctcct 6240 ctgatgctaa gccttacata ctcatacatt tcattggagg gtgccatgag gatggtactt 6300 aggtggtgtg tgttaggctt ttctgcctta tacaagattt gtttaaaaat ggcatgggag 6360 ttggagctga tggttggacg ctggaagacg ttaaaggagg tttcaggtaa gtttacagag 6420 tttttaataa agatttggta ggattctatg agtttgttta ccaactgggc tgttacttga 6480 acgtcgagtg cgcagtaatc aagggtttgc tgaattatgt cgtaagcgcc ctttttttct 6540 tttatccaaa ggtctttgtt aaaggagtac tcctcctgat ccttccagta ctttaagtcc 6600 ggaaagccat cggcgtcttg ttggtaagag ccaagcatgt aaaactcatt cacagcctgg 6660 tagggacagc agcctttgga aaccgttagg ctgtatgctt tggcagcatt tttgaggctt 6720 gtgtgtgtta aagagaaagt gtcacgcacc ataaatttta tgtactggct tttcaagtca 6780 gacgccagta cttggccctg ctcccagtgc ttgtaactct tggcaggcac gtagctgggg 6840 ttaggcagag agtaagtaat atcattgaac agcaaccttc cagctcgcgg catgaagttg 6900 cggcaaattt caaacacagg tggtatttct gcccgcgtgc taaccacctg agctgctagc 6960 aatatttcat caaaccctgt gatgttgtgt cccactgcat acacctcaat aaatcttggg 7020 tttcctttga gcttaaatgt ttttaactga tctggagtaa gctcttcagg gctttctagt 7080 cccaaagtag cggcttcttc ttgaagctgg ggattttggc aaatcacgtg ggaccagagg 7140 ctggcggcaa tatgctgctg caaggtgtcc ctgaactttt tgaatttaac tcctatggct 7200 cttttttctg gagtgatgca atagtacagg gtgtcctgtt ctttaccatt ccaaatttcc 7260 cagttttgtt ctgttgcaag atttttggca acattgacca ggtggttgtc ccccagcagt 7320 tggaaaacta gcataaatgg cacaagttgc tttccaaact tgccgtgcca cgtgtacgtt 7380 tccacatcat atacaataaa cagcctctct gtgttgggcg gggccccaat gggggtgaaa 7440 gatatttttt cccaccagtc tgcgctgtgt gcggccacgt gatggaagta aaagtcccgt 7500 cttctgaggg cgcaggtgtg agtaacttta aaaaaagagc cgcagaactc acacttttgc 7560 atttgagaga tgtcttttat taggaagact ttaccttgct taactaaaaa cttgagtgga 7620 aagctgagca acggaacctt cgcttgaaac tccaccctcc tcactgttcc tcgttggtag 7680 tagcaaatta gactgtaggg tgccgcggcc tcctctatga gggcgggcag gttgtgggca 7740 gtgatgggcc tgtttaagca tgcaggtagg tcatagaggt tagtgttgca aaggttagta 7800 agcgcgctgg ccaagttgga gtgatactta atttccacac actgtccatt atccaacacg 7860 gcggaaatgc ttagtgtggc gcgctggacc accacggcgc ccttgtactt gcgcttgggt 7920 gggattacta tggtggaagt tgcacctgct cgtggtgagc tggcaaaggc ctgtgctgca 7980 tcctgagctg ggaggctagg gccactactc tcctgttgat tgattggatt tggcggttct 8040 gtgaaaagac cactggtccc gtgaccttga acctaaaaga gagttccata gagtcaacct 8100 ctgtatcatt aatggctact tgtctgagaa tttcgctcac gtccccgctg ttttcatgat 8160 aagcaatatc tgccataaac tgctccattt cctcttcctc tagctcccct agcccagccc 8220 tttccacggt ggcagccagg tctcccgaca cgcgggccat aacctggcta aaggcatttt 8280 ctcccatttc attccacacc cgactgtaga ccacctgccc ctcgtggtcc ctagcccgca 8340 tgaccacttg agctagctct aggtcaaccc agcgggagca gggtgggtac agcctaaggt 8400 tgtggtgcag gtagttgagg gtggtggcta cgtgctcagc tacaaaaaaa tacatgaccc 8460 aacggcggat ggtgagctca ttaatgttgc ccagcatttc caggcgctgg atgacctcat 8520 aaaagtcaac tgcaaagtta aaaaactgtt cattgcgagc agacactgtg agttcctctt 8580 gcaggaggcg gatagcttct actatagtgt ctcggacttc ttcttcaaag gaacggcgag 8640 gaggggaggc aggaggagga gaggcctctg cttcgggctc agacagctcc tcagacaccg 8700 gaggcggcgg gggagccctg cgtctgcgcc tgcgcactgg aagcctgtct ataaaccttt 8760 ctatcacctc tccccgcctg cgcctcattt cctctgtaac tgcacgtcca ttttccctgg 8820 gcctaagctc gaaggctccc ccagagagcg gggtgcaggg cgcgggtctg ggcaggctca 8880 gggcgctaat tatgcttttt gttacagatt ggctgggtaa agactgaatt gtttgaaact 8940 gtgacagctc cggatcagtg aatttttcca caaaggcatc aagccacaaa caatcacaag 9000 gtaagcttag catggtgtgc gggttgcggg ggctaactaa aaagttgaaa aaagctgtct 9060 taagcttacg gatggtggtt agaataacta ggtctttgtt gcccgcctgc tggatgcgca 9120 tgcggtctgc cagtccccat gcttcgtttt gacactggga caggtttttg taatggtctt 9180 gcaagagccc ctctacgggc acctgcctgt cgcctgccat gttggtggtg ccatagcccc 9240 taagtggcct aataagggcc aaatcggcca ctacgcgctc tgccaggacc gcttgctgaa 9300 cctgcgccag ggtctctgca aagttttcaa agtcaatgaa cctgtggtaa gctccagtgt 9360 taatggtgta tgagcagttg gccatcaccg tccagtttag tttttgctgc aagggcaggt 9420 gtatctctgt gtactttagc ctgctgtagg acctggtgtc aaagacgtag tcattgcacg 9480 ttctcactag gtactggtag cccaccaaaa ggtggggcgg ggggtaccaa aacagtggcc 9540 agccctcggt agcgggctct cgagggctga ggttcataag catgaggcgg tggtattggt 9600 atatgtactt ggacatccat gccagcccta cagaggtggt ggaagctctg gcgtactctc 9660 gaacccggtt aaaaatgttt ctgagaggcc tgaacacctc aacggtgtat ggggtttgtc 9720 cggttaggcg ggcgcagtcc agcgcgttct gcaaaacaaa caaggagtcg atggcacttt 9780 cattcgctcc attgcagatg catccggtac tgcgacagat gaagcccctg cccgcctctg 9840 ccagcaagct agaagcagag agcgagggtt tggcccgctt acaagggggt gctgctccag 9900 agttgcaccc ccgggtgcag atgaagcaag aggcggcgga ggcttacatc cccgctagca 9960 atgtgtttag ggataatgaa ggagaagaag cagagggctt gcggcactta aagtatgagt 10020 cggggaggct gttgcgtaat gaccacactt ccaagcgggt gctgggcgag cgagactttg 10080 aaaaggaccc ccagaatggc attagcgcgg ccgaggcaca cctaaagtct gcagacttag 10140 tcacggccta tgagcatact gtgaaggcag aggttaattt tcagaccacc tttaacaaca 10200 atgtgcgtac cctcatcgcc agggaagagg tggtcattgg cctgatgcac ttgtgggact 10260 ttgtggaggc ttttctagaa aacccagtga gcaaggccct cactgcccag ctctttctta 10320 tagtgcaaca ctgcagggac gagggggtgc tgcgggaaag cttgctaaac attgccgagc 10380 ctgagagccg gtggctggtg gacctgttaa acctgttaca gactattgtg gtgcaagagc 10440 gaggcttggc agtcggtgaa aaggtagcgg ctataaacta ctctgtgatt actctaagta 10500 aacactatgc aagaaagata ttcaactctg tgtttgtgcc catcgacaag gaagccaaga 10560 ttaacacatt ttacatgcgc actgttgtta agctgctggt gctgagtgat gaccttggca 10620 tgtaccgcaa tgagcggatt gagagggcag tgtctggtgc caggcagcgg gaattaaatg 10680 acagggaact tatgcaccga ctgcggcaag ccttggcttc taacggactg ccggagctag 10740 agggcgaaga cagtgctaat attagatcca aagaggagtg gggggtcgga gcgggcggag 10800 gcgttgcgag cgcgcgctac ccccacctgc tagattatga ggaagaggag aaccctgatg 10860 gctccgtctc atttcaacag catgagcgcg gcgctcaacc ccatgaaaat ggcggccatg 10920 cagagtcagc ctacagccga cgacagttgg gccgcttcta ttagtcgcat tatgtcccta 10980 acggcggggg atcggcacaa gttttcatcc cagccttttg ctaacaggct ggatgctata 11040 ttagaggccg tggtgccttc ccgcaaagac cctacccatg aaaaagtgct aactatagta 11100 aatgccctaa tagaaaatgg tgccattagg cgggacgagg gggcgggggt gtatgacgcc 11160 ttgttgcacc gcgtgtccaa gtacaacagc ttgaatgccc aaagcaacct agagaggttg 11220 gctggagacg tgagggaggc ggtagcgcag caagtgcgca tcgccacggg caacctgggg 11280 tccctaaccg cacttaatgg ctttttggca cgcctgcccg ccaatgtgga gcggggccag 11340 gagaactaca ctggctttgt gtcagccctt aagctgctgg taagcgaggt gcccagtaca 11400 gaggtgtacc agtctgggcc tcactacttt ttgcaaagca gtcgcaacgg cacccagaca 11460 gttaacctta ccaatgcctt tgagaaccta aagccccttt ggggagttaa ggccccccact 11520 atggaaaggc tcagcatctc agccctgctc acccccaaca ctagactgct gcttttgcta 11580 gtgtctccct ttacagacag tgttagcatt agcagggact cctacttggg ttacctgctt 11640 accctgtaca gggaagcctt gggccgcaat catttggatg agcgtactct tgaagaggtc 11700 acagaggtca gcagagccat gggcagtgag aacataaaca acttgcaagc cacccttaac 11760 tttttgctaa ccaatagaca aaaaagaatt cccaaagact actccctaac cccagaggag 11820 gagaggatag tgaggtttgt acagcaggcc gtgagcttgc gcatgatgca ggaaaacttg 11880 agccccactg aagctttgga cgtaaccgcg gccaacatgg agcccagctt ctatgccagc 11940 aacagggatt ttattaataa gcttatggac tactttcacc gcgctgcggc tatggctcca 12000 gactactttt tgggggcagt gatgaatccc cgctggctcc cccctgaggg cttctttact 12060 ggggtttttg actttcctga aagggacagc tacacctggg atggactgga tagctctttg 12120 gagctgacgc gccaggatgc catgcggttt ctggaagaca agtttatgga cgatgaccag 12180 aggacagagt ctcgcagcct cagcagagtt accaccccag ctagctctag gcggtcctcg 12240 gtgaccatgc cacctggggg ctttatgggc atgataaata acaacaaaaa cgataacagc 12300 ctcagggaga tagatgggtt agcagacaag ctggctcggt ggaaaacata caagcgtgag 12360 tcagaggagg ctcgtgtgtc cctgcctgcg gtagtaaggc ctaagcagta caggccccgc 12420 tcccctatct ccagcgatga cagtgatgat gggatgagca ggccagaccc gtttctcaag 12480 tttgagggga gtggaaaccc ctttgcccac ctacgcccca agctggggcg tggcctgtaa 12540 agtgaataaa atactcacca gagacatgac gctagcgttt ttttatagca atggagtttt 12600 cgtcgtctcc tccccccgtct tatgaaacag tgatggcaca agtgccttcg atcctggcgc 12660 cgctggtacc cccgcggtac aaaggggcta cagaaggaag aaacagtatc cgttattccc 12720 agctgccgcc tctgtttgac actacaaagc tgtacctcat tgacaacaag tcttcagaca 12780 ttcaggccct caattaccaa aatgaccaca gcaacttttt aaccactgtg gtgcaaaatg 12840 ctaattacac gcccatggag gctagcaccc agtccatcca gctggacgag cggtcgcgct 12900 ggggtgggga atttaagtcc attttacaca tgaacatgcc taatgtgaca gagtacatgt 12960 ttagcaatag ctttaaggca taccttcccg ccacagcaga caactttggc aaggtgctca 13020 cctatgagtg gtacacccta accattcccg agggtaacta ttctgaggtt atgcttttgg 13080 acctgctgaa caacgcggtg gtggagaact atctggccca cgggcgccag cataatgtta 13140 gggaggagga catgggcctc aagtttgaca ccagaaactt tcacctgggc tttgatcccg 13200 agaccaagct ggtcatgccg ggcttctaca ctaacgaggc cttccaccct gacatagtgc 13260 tgagtccagg ctgtgcggtg gactttaccc acagcaggct caacaacttt ttaggcatta 13320 gaaaaaggct accataccag gagggatttg tgattacctg ggaggacctg cagggtggca 13380 acatccagc cctgttagac ctggaaaatt acaacccaga cattcccggc gccgatatca 13440 caccactgat gtatgactcc aagggaaggc cttaccatgt tggtgaggat cccagcgcgg 13500 gcagtacctt cacctggtac cgcagctggt ttgtggccta taactacggt cctgccgatg 13560 gcatcaagag caaaactgta ctggtggccc ctgacattac ttgcggagtg gagcaaattt 13620 actggagcct gccagatatg gcggtggacc cagtaacttt tacctctagc cacaatccca 13680 gcagctaccc agtggtgggc acggagctgc taccattact gccaagaagc ttttacaatg 13740 gatcttcagt gtacagccag ctgttgcaag aaagcaccag tcaaactcaa gtgtttaacc 13800 gctttccgga gaatgcaatt cttaaaaggc cccctgctcc caccattatc agcatcagcg 13860 agaacgtgcc cgccctcagt aaccatggca ccctgcccct aaaaaacaac attcccgggg 13920 tgcagcgggt gaccattacg gatgcaagaa gaagagtgtg tccctacgtg tacaaaagct 13980 tgggggtcgt tgtgcctcgc gtgctgtcca gtaaaacctt ctagtgacaa actttttctt 14040 ttcattagca aagccatggc catcttaata tcaccgacta acaacaccgg ctggggccta 14100 ggcactcaca agctctttgg aggagccaaa caaaaatcag atcagcatcc cgtttatgtc 14160 caagctcact accgggcctc gtggggcagc aaagggcgcc gccgccgtca aggccgagcc 14220 cgcggggccc ctcttgaccc taaaactgaa gcagaaatgg ttgccaccat cgacgaggtg 14280 gcgcgcaacg gtccccccggc ggcgcgcttg gtgctggagg ctgcccggag agtgggggct 14340 tacaacctcc gtcgagcgcg caagctaacc cctgcggggc gtgccatgat ggccatgcgc 14400 gcgcgtcaga tggtaaagca ggctaagaaa aggaagcgca gggttcgctt tagacaataa 14460 agtctgtcct cacatacaca catacccgag tgatggccta tttttttact gaagcctcat 14520 cgccatggca gcaatcagtc gcgccatcaa gcaggaactg ctggaagacc tcaaacctga 14580 aatctatctg ccacccaagt ccaccaaacg gagaactaaa gttaaaacag aggaaaaagt 14640 tgatgtgaaa accctagtca aggccaagag cagaaagcgc agggctgcca aggatgagct 14700 ggaagaggat gtagagtttg ttaggcgctt tgctccccga cgtccctacc agtggcgggg 14760 ccggcgcgtg cgcgctctgc ctaggcccgg ggtgcccgtg gtgtttactc ccggccagag 14820 gtcgggcacc gcctcaaaaa ggtcttatga cgaagtgtac gcagatgagg atgtactgga 14880 ccaggcggga aacatgatta atgagtttgc ttatggcaag cgagtcaggg tgcttaccca 14940 caaaaacccc actccctcac aagtgcccat caccccccag gagcccgtgg cgcgtcctgg 15000 cgaggcacac ctgctaccca cagtgcaagt gctggctccg cgaggctcta ggcgggaaac 15060 catgttgccc gtaaccaagt ccgagggggg agacgtgaaa gtggaaaaca agggctttga 15120 gcaaatcacc cccgggctgg gggtgcagac tgtagacatc aaagtgcccg tgaagcgcaa 15180 aggcgatgca gaggatgaaa taataaagag ggttaagatg gagcttgaac catatgagac 15240 cactatgaaa atggagtact ctgaggagcc ccaggttgag gcttttgaca cagggataga 15300 gcccagctct ttttttgagg tgcgctctca agcgcgtccc atcgctgtgg ctaggaagcg 15360 ccgcaccgca gctgccagcg cccccgctgt agaggtgatg gaggttcagc aaagcaaccc 15420 tgccacccca gctaccgctg ccgccagaac cgcgaccgcg ctgggcccgc gtttggctcg 15480 tcggccttca agatgggggc ccgccaatgc catctttcca gattacaaat accaccccag 15540 catcacggct aggaaaataa ggggccccaa gcctacaggc aagatcagtc gctgggggcc 15600 tgccaacagc atcctgcctg aggtgcggct gcaccccagc atggtgtctg cggtgaccag 15660 agctgcccct cgcagagtca ccaaaacccg gagacgccgg cgtgccagaa cccgccaggc 15720 ttttgtgctg cccgcgcgca ccaaaacggg cgcgctgctt tcccagaacg tgcgctacca 15780 cccttccatc tccttgctcc gccgagccta atctgtatgt tttgctttgc gcaaggatgg 15840 ccggcaggaa tgtcaccctt cgtctgagag tcccagtgcg caccaaaatt acaggcgcag 15900 gtcgtcggcg cggacggcgc ccacgcggca tacgctgcgg gcgcatgaga ggaggctttt 15960 tgccagccct catccctttg atagctgccg ccattggggc agtaccagga atagcttcgg 16020 tagcgctcca agctgcccgg cactagcttt ttacagattg actcactccc atttttcttg 16080 tgccagaaga agacaaaatg gatgctgtca atttttccat cttggcacca cgctacgggg 16140 cccaccctat gatgagtggt tggtctggca tcggcaccag cggcatgaac ggcggggcct 16200 ttaactgggg gggcatttgg agcggcatca aaaactttgg cagtaatgta aaaagctggg 16260 gcagcaaggc ttggaacagc caaacgggca agctgctcag gcaaaagctt aatgacacca 16320 aagttagaga aaagctggtg gagggcattt ccaccggggt gcacggggca cttgatatcg 16380 ccaaccagga gttagcaaag caaatagagc ggcgtttgga gcgtcagcag cccctagagc 16440 cggaggtgga ggtagagggag gaagtagtgg acataaaacc cgaagcgcag gccccgctgg 16500 tggtacaaat acccaaaaag cgcccccgcg acgaagactt gctcataact gcggacgagc 16560 ctccctccta tgaagagtcc atcaaaacca tggcacccct gatgcccatg acccgcccgc 16620 acccttccat ggccaagcct gttttggtag accgtcccac caccttggag ctaaagccct 16680 ctgaccagcc gccagcctac tccccccccgg cgcccagcgc cgtcagggtc acagtgccct 16740 ctaacattcc cgtggtcact cctgccagaa gcagaggctg gcaaggaact ctggccaaca 16800 ttgtaggagt gggtctaagt aacgtgaagc gccgtagatg cttctaactg tgattaaata 16860 tactcaagtg acttgaagtt tgccgaccct gtcttttttt gaccgcgcca gctgagaaga 16920 tggcaacccc gtcgatgctg ccacaatggt cttacatgca cattgctggc caggacgccg 16980 ccgaatactt gtctcccgcc ctggttcagt ttgcccaagc aaccagttct tactttaagt 17040 tggacaacaa gttcagaaac cccactgtgg cccccaccca cgatgtgacc actgagaggt 17100 cgcagcgctt gcagctgcgc tttgtgccag tcatgcaaga ggatggccag tacacttaca 17160 aaacccgctt ccagcttgcg gtgggagaca acagggtgct ggacatggcc agtacttact 17220 tcgatatcag gggtacccta gacagaggcc cctccttcaa gccttacagc ggcaccgcct 17280 acaatgccct cgcccccaag gccggggcta acaactgtct ttttaatgga cagggtgcca 17340 atattaacac tttagcccag gtgccctctg caggtgccat aactgtgaat ggccaagctg 17400 ctgtcacaaa caatacctac cagccagagc cccagctggg ccctgaaagc tgggtcgatg 17460 gcagcctagc agagctggga gatgcgtctg gccgtgccct taaggcttca accccgcgca 17520 tgccttgcta tggttcctat gctcccccca ccaacgaaaa tggaggtcaa gcaactggtc 17580 cagtggaatc cagattttat aaggtgacca ccaacaataa caatgaagca gatgccatgc 17640 tatacactga agatgtgaac ctgcaggccc cagacaccca cctggtgcac caagtaccag 17700 agggtcaggt tacaggggta caagggctgg gccagcaggc tgcacccaac aggccaaact 17760 acataggctt cagggacaac ttcataggcc tcatgtacta caatagtaat ggaaacctag 17820 gggtgctggc gggtcagtca tcccagctca atgccgtggt ggacttgcaa gacagaaaca 17880 cagagctctc ttaccagttg ctgctggatg ccctcacaga caggtcccgc tacttttcca 17940 tgtggaacca ggctgtagat agctatgacc aggatgttag gattattgac aaccatggcg 18000 tggaagatga tatgcccaac tattgctacc cactgagcgg catggggccc ctaacaaaca 18060 tgaccaccat gaaggttaac aaccagaact ttcaggcaga aaatgccaat gtgggaccca 18120 ttcaaaaaat tggttttgga aatgttgagg ctatggaaat caacctcaat gccaacctct 18180 ttaaaagctt cctttactcc aatgtggcct tatacctgcc tgatgccttt aaatacacac 18240 ctgaaaacat tgtggcccct gccaatgcga atacctatgc ttacatgaat gttagattac 18300 ccgccgccaa ccttatagat accttcgtaa atattggcgc cagatggtca ccagatgtaa 18360 tggacactgt taatcctttc aaccaccaca gaaatgcagg actccgctac cgttcacaac 18420 tgcttggcaa tggccgctat tgctcgttcc atattcaggt ccctcaaaaa ttttttgcaa 18480 tcaaaaatct cctcctactg cctggaacgt acacgtacga gtggtctttc agaaaggatg 18540 taaacatgat ccttcaaagc agcttgggca atgacctccg agtggatggg gccaccatca 18600 acattcagag catcaaccta tatgcaagct tttttccaat ggcacacaac actgcctcca 18660 ctctggaagc catgctgcgc aacgatgtaa atgaccagtc ctttgcagac tacctgtctt 18720 ctgccaacat gctttatccc atccctgcca acactactaa cctgccaatt tccattcccg 18780 ctagaaactg ggcgggattt agagggtgga gctttaccag aattaagcaa cgagaaactc 18840 ctgccctggg ctcgccttat gatccctact tcacttactc gggcagtatt ccatatctgg 18900 atgcaacttt ttacctcagc cacaccttta gaagagtttc catcatgttt gactcttcag 18960 tgtcttggcc tggcaatgac aggctgctta cccccaatga gtttgagatt aaaaggtatg 19020 tagacggtga aggttacaat gtggcccagt ccaacatgac aaaagactgg tttatggttc 19080 aaatgctagc ccactacaac atcggctacc aaggctacca cctgccagaa agctacaagg 19140 acagaatgta ctccttccta agaaactttg agcccatgtg cagacagttg gtggacgtgg 19200 ccaactatgc tgcgtaccag ccagttaccg tgggccacca gcataacaat tctggttatg 19260 ctagcgccct ttcggccttt aacccacgtg agggacaccc atacccagca aactggcctt 19320 acccactcat tggagccaat gcagtaccca ctgtcaccca gaaaaagttt ctctgcgaca 19380 ggtccctgtg gcgcatccca ttctcctcca actttatgtc tatgggaacc cttactgacc 19440 tgggccaaaa cctgctgtac tctaactccg cccacgccct tgacatgact tttgaggttg 19500 atgccatgaa tgagcccact ctgttgtacg ttttgtttga agtgttcgac gtggcacgcg 19560 tccatcagcc ccaccggggg gtgattgagg tagtgtacct cagaactccc ttctccgccg 1920 gcaacgccac cacctaagct aatatggcgg aaggagggtc ctcagaagaa gagctgcgag 1980 ccatagtaca taacttaggc gtgtctccat ttttcttggg cacctttgac aagaggtttc 197440 cgggtttcat ctcttcacaa agaatggcct gcgccatagt aaacaccgca ggccgagaaa 1800 cggggggcgt gcactggctg gccatggcct ggaacccccg ctcaaaaacg ttttacatgt 19860 ttgacccctt tggattttca gacactaagc tcaagcaagt gtacagcttt gagtatgagg 19920 gcctactcag gcgcagcgcc atagcctcca gcccggatag gtgtgttact ttggccaaaa 19980 gcaatgaaac cattcagggt cccaatagcg ccgcctgtgg actgttttgt tgtatgtttt 20040 tgcacgcctt tgtcaactgg ccggacgacc cctttgacca caatcccacc atgggacccc 20100 tcaagagcgt gcctaactac aaactcaatg atcccacagt gcagtatgtg ctctggggaa 20160 accaagaaaa gctgtataag tttttggaaa aacactctgc ttactttcgc acccacgccg 20220 ctgcaattaa agccaggact gcttttaata aattgaaaca ataaaccgtt tattcgaaaa 20280 atataaatgg tttacagtgt gattattaaa aaaggacaag gtcatcgtca tgttggccct 20340 ggggcaaaat ggtattttgg tacctgtgtt catcggacca gtggaactct gggaatttaa 20400 tggggggcct ttcgcccact gtggacgtcc atatttgctt agcaatttgg acacatgaca 20460 ccaggtccac gctggaaatc ttaaaatcac aatttttttg gggggcggcc ttgctgttgc 20520 ggtacactgg attgcagcat tgaaacacta gcatggcggg gttattaagg gtggctagca 20580 ttttttggtc atccacttca cttttgtcaa tgtgactagc gcttgcaaga gagaatgggg 20640 taatcttgca agtttgtctg cccagcaaag gaaggttact gttccaattg cactcacact 20700 tgagtggcat tagcaagtga ctctcagcat ttggcataga ggggtagcaa gccttaagaa 20760 aagccatgat ctgctgaaag gccataatag ctttgggtcc atcggtataa aacataccgc 20820 aagaggcccc gctaaagttg cccccgctca tattaacatc catgctgcag cagaaagcat 20880 cctcattctt aaactgtact acatccttc cccaccggtt agaaacaatt ttggtttttt 20940 ctgggttttc cttaagggcc cgctgggcat tttcactgtt aacatccatt tctactagct 21000 gctctttttg gatcatgggc gacccgtgca gacacataag gctgctttgg catccatgtt 21060 gccacaccac acacccgctg gggttaaccc caggggcaag ctctgctgct ttgattacaa 21120 aatcaagcag caggcgccct gccacgtgca aaaaagtttt ctggttggta aaggtgtaag 21180 taacgtgcgt tttagaggtg gtaatgtaag cctggactgc ttttttaaag cactccagtg 21240 tgccctcgtc tggaagcaag gttagcttgc tgtggtctac ttggtacgcg gtgagaacct 21300 taagggcaag ctccatggcg gactgccact tgtgctcact agctttaaaa aacttagaag 21360 ccactggccc cgagggcgcg ttgggcttgc tccaaacgga cttttgcttg ggtgggataa 21420 ataccacatt tccctccttg tcagtgtcca ccgtttggcc acaaaaggtc tgcactacaa 21480 tgctcccggg atccccccgc gccttgaggg tggcggtggc ttgacgggct atttcgacca 21540 tggcttggtg gccccttggc tcctcaacgg ctttccgctg gcgcttcttt gggggcgccg 21600 taggcggtgc cacaggaacc tcttcatcgc tggagctttc agagattgca acaaccttct 21660 tgtggctcat ctttttccta gatgtcagaa gagcccgtca gtgggactac cgtggagaca 21720 aaggaggaca ttcacacacc tccaaactcc cctgtcctcg agacaatctc tctaagcccc 21780 gaacccgagg ctgaggcctg cccaaataca gacaaatacc taagcaccaa tttgctttgc 21840 aaacacctgc aacgtcaaag cgcaattgtt ctggacagta tcaaagatca tctacaagtg 21900 cccaccagtg tatcagagct aagctgcgcc tacgaacgaa gcttattctc cccaaacacc 21960 ccaccaaggc aacaaagcaa tggaacgtgt gaagcgaatc ctaaacttaa cttttaccct 22020 acttttttgg tgcctgaaac tttggcaact tatcacatct tttttgttaa tcaaaagata 22080 ccagtatctt gcagagctaa cagagctaaa gcagacagag ctcttgcttt gcaagaggga 22140 gattgcttac ctgactatga aacaatggac acagtcagcc gggtgtttga gggtttaggc 22200 ggtgaggtgg ttgcggatca cgcgctgcaa aataatgact ctgtattagt ggagcttaag 22260 gaagacaacc cccgtttggc tgtggtcaaa agaaacctca gcgtgtccca ctttgcctac 22320 cccgccgtgc acctgcctcc aaaagtcata accactgtca tggacaacct gctagttaaa 22380 aggtccaccc ccagcgccga cgtctcagag ctagatcccg aggggggcca agaggtggtg 22440 tctgacaccg agctaagtaa gtggctaaac acctctgacc ccgaggcctt agagaagcag 22500 cgcaagacag taatgggaag cgtgctggtc actgtagtgc ttgagtgcat gcaaaggctt 22560 ttcacctcag aagaaatggt gaaaaaaatt ggggaggccc tccactacac ctttaggcac 22620 ggctacgtgt ctctggcttg caagatttct aatgtagagc tgaccaacgt ggtcacctat 22680 atgggcatcc tgcatgaaaa ccgcctgggt caaaccaccc tgcatcacac agtgcagggc 22740 gaggcccgtc gagactacat tagggactcc atcttcctca tcctaatcca tgcctggcaa 22800 accgcgatgg gcatctggca gcagtgcctc gaagaggaaa accttaaaga actagccaaa 22860 ctgctgcaaa ggattaaaaa gccattgtac actgaaacct cccagcgcct tatgggcaag 22920 ctcctggcaa acgtagtctt cccacccaaa ctgctacaaa ccttccacaa gggccttcct 22980 gatattgtta gccaaagcat gatgcaaaac tttaggtctt ttattttgga aaggtccggc 23040 atcctgccct ccatgacctg cgcgctgccc acagatttg taccaattca ctttaaagag 23100 tgccccccca cgatgtggcc ctacacctac ttgctcaaat tagcaaactt ctttatgtat 23160 cacaatgacc tttgctatga tgtgggaggc gagggcctgc tagagcacta ctgccggtgc 23220 aacctctgca ccccccaccg ttgcctagcc accaaccccg ccatgcttaa tgaaactcag 23280 ctaataggta cctttgatat tcggggtccc ggcggagaaa acggagcaga gtcttcctct 23340 ggccttaagc tcaccgctgg aatgtggact tccgcgtttc tgcgaaaatt tgaaagctct 23400 gattatcacg cccacaaaat tcagttttat gaaaaccaat caaagccccc ctcagtggag 23460 cccactccct gcgtcatcac ccaaagtagc attttggccc aattgcatga cattaaaaaa 23520 gcaagggagg agtttctgct aaaaagcggg cagggtcagt atctagaccc tcagactggg 23580 gagccgctca acgctgcgga tccttccgta gaaagcagcc atgagttcca aggagatgga 23640 agacacagaa accctaagcg tggaagaaat ttccggcagc gaggaggacc tagaaagcct 23700 cccagagtcc atgccggagc agagcgagat gccagaggaa cgagctccta gatgggacca 23760 aaaagagaag cctctgggta agcagccccg caactaccac tcctggagag cgcataagtt 23820 taagatctt agctgcctgg gcgccagtgg gaacaatgtg gcctttacca gaagatacat 23880 gctctttcgc gagggagtta accttcccaa caacatcatt cactactata actctcgcta 23940 ccgcagcaaa acagaagctc aagcgcagac cgaccccact aaagagccca aagccagccg 24000 ccggtgagga caaccaccag acaaggcagc tgcgcaaccg catttttcca acactgtacg 24060 ccatcttcca acaaagcaga ggctccccca cggcattaaa aataaaaaac agatccctca 24120 gatccctcct aagaagctgt ctctaccaca agtcagagga gcagctgctg cgcacgcgaa 24180 acgacgccga agctttgctc aacaaatact gtcaaggact cgagtccggc acagactgag 24240 caatgtctaa agaaatacca accccttata tgtggagcta ccaaccgcaa acggggcacg 24300 ccgccggcgc ctcccaggac tactccaccc aaatgaattg gtttagtgct gggccatcaa 24360 tgattagtca agtttatggc attagagact tgcgcaacaa agttttgata acccaggcag 24420 aaataaccaa aactcccaga acaataatgg atccgccaat ttggcctgct gccatgcttg 24480 ttcaggaagc cgccccaccc aaaacggtca ctctgcccag aaaccacacc ctagaacagg 24540 ctatgaccaa ctctggggca cagctagcgg gaggacgaca gctgtgcccc tcccaaatag 24600 gtataaaaag cccagtgctg gctggcacgg gcattcagct tagcgaagac atccccagcg 24660 cctcctggat caggcccgac ggcatattcc agctaggagg ggggtcccgc tcgtccttca 24720 gcccaacgca agcattcctc accctgcaac aggcatcctc gacgccgcgc gcaggaggcg 24780 tgggcaccta ccagtttgtg cgcgaatttg tgccagaggt ataccttaac cctttttcag 24840 gaccaccgga cacctttcct gaccagttca ttcctaacta cgacattgta accaactctg 24900 tcgatggcta tgactaagga gagcatggac caggtggagg tgaactgcct gtgtgctcag 24960 catgcccaaa cctgcacgcg ccctcgctgc tttgcaaaag agggtttatg tgctaactgg 25020 ttttacaacc cagcacttgc ctttgaaggg tttgatattc cagactctta ccaagaggga 25080 cacggtgtgg acatagaagt taagtgttcc caccactcca gcaaactgtg tcacaatggc 25140 catgatatga tctgctcata ctctcgcctg ggatcccaca ttaacataag atgtatttgc 25200 aacaagccgc ggccccacat gagcctcatt gaggcagcct gttctatgta taaccttaac 25260 tagataatat tattaaactt gttttacagc taccaccata atgcgcttca gcttcttcat 25320 cgccgccgtc cttttctgca ccacaggggc tagcaatgac attgtgactt gctgcgccca 25380 cacaccttgc ctcctacacc tagaagtggg cttgggggcc aatgtcagtt ggataaactc 25440 tgacacaggc caggccccgc tttgcctctc caatggcatg tgcaacgcta cccagcaagg 25500 cctgcagttt tctgcaaact tttctgagga tggcctgtac atcgccctca ttaaggagag 25560 caactacgag ggcgctgagc actactacct tgtttatatt tatggagact gctaccaaac 25620 tgcaaatgag tctgcccacg ggcctatttc cagacccctc aacgagatgc ctcttcacag 25680 cgtaaccata aatgcttccc tcttctatcc cgcctttctg gagctgcccc cacagtacag 25740 caatgacctt agcaatgtgc gctggtataa agtagacccc agcggcttcc aagcccaaaa 25800 aatctctaaa gtcagaagcg gaggcagaaa agagaacctg catcccaact gggccttggt 25860 tacctatact ggagaccttc ttgtcttgca tgtctcgcca aacacccttg gactgtggct 25920 ggcagccgtg cagcatcgcg ggggacgcac taatttcatt accttcaaca taactgtacc 25980 caactggcaa caaaatctag tagccatatt taatcaacac gagcccccaa aaatgggcaa 26040 taattatgag gacagtttta tggaatggac tctgtttaaa aagctcaaaa aaggcttatt 26100 tagagtaact tgcagagcca agtcaatatt tccagagtgc accctcaaca tcacccgcga 26160 cggaactttc ctgcttattg gggaaagcaa gaagaccccc tatgtcatcc tgctgccctt 26220 ttttgcaaac cccaaagaag acactccaat tttaatggcc cttagccact ccgtgcccgt 26280 tgccatacct aacactgcaa tgcctgtata tatttccatc atgtttttta ttgtagccat 26340 gctagccacc ctcagccttc taatgggact aaacaacaaa atcaggtcca tgtagcttgt 26400 caaataaact tacctaattt ttgctaagac gtctgggtcc tgcgtttcta tgtccaccaa 26460 agtcccctct tcccagcttt ggtacttcca cttgtgtgtg cgagccagct tgcggatgtg 26520 cttgaaagat aatgtggtct ctcccaacag cttcccgttc accagcacca gggccatgaa 26580 gcggacacga agagctctac ctgcaaatta tgaccctgtg tatccatacg acgcccccgg 26640 gtcttccaca caacccccat tttttaataa caagcaaggt ctcactgagt cacccccagg 26700 aaccctggct gtcaatgttt cccctccact aaccttttct acgttaggtg ccattaaact 26760 ttccacaggt cccggactca ccctcaacga gggcaagtta caagccagct tagggcccgg 26820 cctcatcaca aataccgagg gccaaatcac tgttgaaaat gtcaacaagg tcctgtcttt 26880 tacctcccca ttacataaaa ctgaaaacac tgtatcccta gcgctaggag acgggttaga 26940 agatgaaaat ggcaccctta aagtaacctt tcctactccc cctcccccgc tacaattctc 27000 ccctcccctc acagaaacag gtggtactat ttccttgccc ctgcaagact ccatgcaagt 27060 gacaaatgga aaactgggcg ttaagcctac cacctacgca cctcccttga aaaaaactga 27120 ccagcaagtt agcctccaag taggctcggg tctcaccgtg attaacgaac agttgcaagc 27180 tgtccagcct cccgcaacca cctacaacga gcctctttcc aaaactgaca attctgtttc 27240 tctgcaagta ggtgccggcc ttgccgtgca gagcggagct ttggtggcaa cccctccccc 27300 gcctctcacc tttacatcac ccctagaaaa aaatgaaaac acagtgtcgc tacaagtagg 27360 cgcgggcttg tctgtacaaa acaacgccct agtagccaca cctcccccac ccttaacctt 27420 tgcctatccc ttagtaaaaa atgacaacca tgtagctcta agtgccggaa gtggtttaag 27480 aatatctgga ggcagcctca cggtggccac tggacctggc ctttcccatc aaaatggaac 27540 aataggggct gtggtaggtg caggcctcaa gtttgaaaac aatgccattc ttgcaaaact 27600 aggcaacggt ctaaccatta gagatggtgc tattgaagca acccaacccc cagctgcccc 27660 cataactctg tggactgggc ctgaccctag cattaatggc tttattaatg acactccagt 27720 aattaggtgc tttatatgcc taaccagaga cagcaactta gtcacagtaa atgctagctt 27780 tgtgggagag ggggggtatc gaatagtcag ccctacccag tcacaattta gcctaattat 27840 ggagtttgat cagtttgggc agcttatgtc cacaggaaac attaactcca ccactacttg 27900 gggagaaaag ccctggggca ataatactgt tcagccacgc ccaagccaca cctggaaact 27960 gtgcatgcct aatagagaag tttactccac tcccgccgcc accatctccc gttgtggact 28020 ggacagcatt gcagtcgacg gtgcccccag cagaagtatc gactgcatgc taattattaa 28080 caaaccaaaa ggcgttgcca cttacaccct tacctttagg tttttaaact ttaacagact 28140 aagcggaggt accctgttta aaactgatgt cttaaccttt acctatgtag gcgaaaatca 28200 ataaaaccat aaaagaaata agtttaaaag ctttattttt catacacgcg agcggtaagg 28260 ctgccgcctt caggaaaagt tactttgtaa acagttcttt cacaacagca caaaacatag 28320 gtattagtta acagttcatt tgggctataa taatatacat tttcttgggt ggcaaagcaa 28380 gggtcggtaa tctcaacaaa accatcaact ggaatgcaag aatagtccag cacggtgggt 28440 tcaatctaaa aatgaagaaa cgcgttgagg ttcactaagc acaggtttcg aatctgtcgg 28500 cagcgtccat acatcatagc ttgtctcaaa gcagattgtc ttctttcctc tgccttggaa 28560 gtagtttggt gaagcactac aggtgtcttt tcaacctctt tcagcacccg ctctattaca 28620 gatctcaccc acacagcaca gtttttaaga gaacaatagt tttgaaggct gcaagattta 28680 cacttaagca ccagccagta attataagtg cttttaagaa ctacccctag ctcagggtta 28740 atgcaccttt taatggcctc catgcaggct ttatggacag ttctaaaaaa agacagtcta 28800 aaataaatgt agtgagtgtt tctaaatata atactcccca catagttaat ctcatcaggc 28860 ctgctagaat ttacaaactc tcgataccac atatactttt tattcatagc cccaccctta 28920 acaaagtcct caatcacttt ctgaaccaca tgcttgctag ccatgcattg taaagacaag 28980 ctgttagaac agtgacagtg taattgccac gtttgagcct ctgccaggca gcagtgctta 29040 gttactatca actcaatacc cgcattgcat gtaaaccccc caaatagcag tttttcatgc 29100 ctgtgtagca catcatccca caaaatagga atttcatagc ataaagcaaa gcaattacaa 29160 tatttaggaa ctctcaccac agcagtcacg tggcatgttg tctcagcagt gcagttgcct 29220 tccatcctac aattatgaac aaaaactaaa cacttctaac aaagatacag tgacaatctc 29280 ccttcctcta aaagcattgt ttacattagg gtgattatta acaacgtcag aaatttcttt 29340 aattaaagtg cctttaaaat gtgcaagagc atcatcatac tcaaaaccaa gctgagagta 29400 aaaaaccacc ttaaaagtaa tcccaggctt gtttttatca acagccttaa acagctttc 29460 acaaaatata gaagcagtaa catcatcaat ggtgtcgaag agaaactcca taggagactc 29520 cagcattgat ccaagctctc taacaaaatc ttcctcaaaa tgaataatgc cctttacaca 29580 aacgcggggc agacgatggt gggccatcgc gtcaacctga aacacatttt acagtaaaca 29640 aagctagctc cgcagtggta aagtcatgcc catgggtgat accaaaatcc ttaaaaaagc 29700 tatctaagta gttggtcatc ccctcagtta aaaagttttg cagctgggtg gtgcatgcca 29760 catagtgcca gcttatagct acaaagacct ccatcccctc cttaacaggc agctcttgca 29820 cacacgcagt aactatccac cgcttaagaa aagctttaag cccagcgcac ataacagctc 29880 caatgttttt atccaaggag agcaaaattt cagcaagcgc aggctccaca gtaatagtaa 29940 agcagaggca tttcagacga ggctcactat ctgcagtcgc catttatgag gtctgcaata 30000 aaaaacaact catcagcagc tgaaaaagtg cactttgacc tcattaagcc actgcatatg 30060 caagtcctca tctatgccgc aacccagacc ctcaatccag ccccgaatgt acactttaat 30120 aagagattca acctcttctt ttagtaaagt gcacatgctg tttggactag tatacacaat 30180 agaagtcaca atgaggggcc cgctgtgact ggaaagcctg cgcacagccc gaaggttaaa 30240 aatggactgt aacagcattg aaaccccgcg acacaggtca gtctcgcggt cttgatctct 30300 tattatagcg accaaatggt ccttcaaagt aatgttgcac tcatagaagt aggcagctcc 30360 ggaagccatt ctgcaaaata acaaaacacc actaagcata gcaccgtcac caagcatgaa 30420 aacaggtaaa aacaaaagca acacttactt attcagcagt cacaagaatg ttgggctccc 30480 aagtgacaga caagcctaat gcaaggtggg cacagtcctc cggaataagt tgacaaaagt 30540 cacgccgcaa ggcttcctga agagaaacgg cggtagcctg gatatctgca acggagccaa 30600 aaccttcagt gtcacttcca ataaacagat aaaactctaa atagtcccca cttaaaaccg 30660 aaacagccgc agcaaaggta ggacacggtc gcacttcctg agctctaata aggctaaaca 30720 ccacacggcg cagttcagaa ggcaaaaagt ctgtaagctc tagctgggca cacacactct 30780 ccaccagact cttgtgaagt cttagacaaa aacatgctcc catagacact cctaaagctg 30840 ccatagtact cacggacggc tggctgtcag aaaagagcta tgagggtgaa atgccaagca 30900 cagcgtttat atagtcctca aagtagggcg tgtggaaaac gaaaaggaat ataacggggc 30960 gtttgaggaa gtggtgccaa gtacagtcat aaattgtggg cgcgtggtaa atgttaagtg 31020 cagtttccct ttggcggttg gcccggaaag ttcacaaaaa gtacagcacg t 31071 <210> 2 <211> 31053 <212> DNA <213> Artificial Sequence <220> <223> APQA1701-40P nucleotide seq. <400> 2 catcatcaat aatatacagg acaaagaggt gtggcttaaa tttgggcgtt gcaaggggcg 60 gggtcatggg acggtcaggt tcaggtcacg ccctggtcag ggtgtttcca cgggaatgtc 120 cagtgacgtc aaaggcgtgg ttttacgaca gggcgagttc cgcggacttt tggctggcgc 180 cccgggtttc tgggcgttta ttgattttgc ggtttaccgg gtggtgcttt taccactgtt 240 tgcggaagat ttagttgttt atggagctgg tttcggtgcc agtttcccca cagctaatgt 300 gaaagtttat gacaatataa cagaaacact ctgttctcag tttacagcac cccacccggt 360 ggttttttgc cacgcctttg ggttaatttt atttccctat acgcagcctt aaattctcag 420 tgcagacgaa agaggactac tcttgagtgc gcagcgagaa gagttttctc ttcactctgt 480 ctgatatatt ttctgaaaaa tgaaatatac tattgtgccg gcgccgcgca acctccatga 540 ttatgtttta gagctactgg aagagtggca gccggactgc cttgactgtg agtatcctca 600 tggcggcccc tcgccgccta ctctgcacga tctttttgat gttgagctgg agacttctca 660 cagccctctt gtgggcctgt gtgattcttg tgcggaggct gacactgatt cgagtgcgag 720 cactgaggct gattctggct ttagtccttt atccactccg ccggtttcac ctcttccacc 780 gcatcccacc tctcctgcta gcatttctga cgacatgttg ctgtgcttag aggaaatgcc 840 cacctttgat gacgaggacg aggttcgaag tgcggcgacc acctttgagc ggtgggaaaa 900 cacttttgac ccccatgtgg gtcctatttt tggctgtttg cgctgtgctt tttatcaaga 960 gcaggatgag aatgcacttt gtgggctttg ctatctaaag gcccttgctg aaggtaagtt 1020 ttaagttaat taaatgtttg ggcaggttaa atgttttagg tgtgtattga ttttaaattt 1080 tgctttttag tgccttttgc tttgcctgta cgttcagagc ccgcttcggc tggagctgag 1140 gaggaagatg atgaagttat ttttgtgtct gccaaacctg ggggcagaaa gaggtcagcg 1200 gctactccct gtgagccaga tggggtcagc aaacgccctt gcgtgccaga gcctgagcaa 1260 acagaacctt tggatttgtc tttgaagcca cgcccgcact aatctgtttg agtacaaagc 1320 aataaagtaa tcttgtttaa caagtttgcc tacatttgtg gttttacggg gcggggcgag 1380 tagtatataa tgccaaaagc cagtgcctgc ttcattaagc ttctagactg agctaagagc 1440 aggtagtatg gaccctctca agatttgtga aaactacctt acttttagag ctataattag 1500 gggaagtact ttgtcgcctg gattttttag gcggtggtgt tttcctgcct tggctgatgt 1560 ggtgggcaat atagtggaac aggaggaagg caggttttgg caaattttac ctgaaaacca 1620 cactttttgg ggtcttttgc gcaggggctt tactgttgct tcttttactg aaattattac 1680 agcagctcag ttggaaaata gaggtagaca gttggccttt ttagctttta tatcattttt 1740 gctacgcaac tggccttctg actctgtagt gcctgaagct gacagacttg acctggtctg 1800 tgcgccggca tggagccaga ccgccaggtt aatcaacgac ctccaagatt ccgtgctcga 1860 ggagcagggg tccgcggaag aggaagagtg cgaagaagcg cttttagcag gggacagcga 1920 cgacccatta ttcgggtaga tgacttgcag ctgcccgacc ccctgtatgt tatgcaagct 1980 ttgcaacggg accacacttt agaaatgccc agagggcagg tagattttag ctggattgag 2040 gctgaagaga ggcgggtagg tcctacagac gagtggtact ttgaggctgt gaagacttac 2100 aaagctaagc cgggagatga cttgcaaact ataatcaaaa actatgccaa gatttcctta 2160 gaatgtgggg ccgtgtatga aattaattct aagattaggg ttacgggggc ttgctacatt 2220 attggtaatt gtgctgtgct taggcctaac ctgcctgctg gagaagcaat gtttgaggtt 2280 ttgaatgttg attttattcc ctctattggt tttatggaaa ggatagtgtt ttccaatgtt 2340 atttttgatt gcaggaccac cgcgactgta gtgtgttgca ttagtgaaag aaacaccttg 2400 tttcacaatt gtgttttttc tggccctcac atgttatgtt tggaccttag ggcgggggcg 2460 gaggtgaggg gctgtcactt tgtgggggcg gtgtgtgcgt tgcgtagcaa ggggctgtac 2520 agtattcgag tcaaaaatag catttttgaa aagtgtgctt ttggggtggt gaccgggtca 2580 aaggctttta ttagccattg catgtttaag gattgtacct gctctattat gctggggggt 2640 cagggcacta ttgcccatag tcagtttatt gtaactactt ctgctgaggc ccccatgaac 2700 ctgcaactgt gcacttgcga gggtaatgga agtcatgtag ttccattggg gaatattcac 2760 tttgcttctc accgggaagc ttcgtggcct acgttttatg caaacacctt ggttcgggtg 2820 cgcttgtata tgggccggcg ccggggagtt tttcacccca agcagtctac tttgtcaatg 2880 tgtgtaattg cagcccctcg gggggttgtg cagagaattt atttgtttgg tgtgtatgat 2940 gctacttgtg ccattatgca actgggcgag gcaggcaatg ctgctagtga aagactgtgt 3000 acttgcgggt tcagacacag caccccttcc ctgcgggcca cctatgtaac tgacaccagg 3060 attgaccggg agctgaactc tcaagacacg gctgagttct ttagcagtga tgaagataat 3120 ttttaggtga gtagatgggc gtggtttggg ggagtataaa aggggcgcgg tacgtggctg 3180 tgtatttaca gccatggacc ctcaacagaa ggggcttgtg aacacgtgtt ttgtgactac 3240 gcgtattccg tcttgggcag gagcaagaca gaatgtcacc gggtcagatt tagaaggaaa 3300 gcccgtgccc tcagatgtgc tggaaagtgg acgcccgctt gcagccccgc gcatcagaac 3360 tttgtatgag gagcagcagc tgaacatgct tgcagtgaat gttcttttgg atgagctgaa 3420 gatccaggtg gctgccatgc aaaactctgt gactgctatt cagcgagaag taaatgatct 3480 aaagcaacga atcgcccgag attaatgtaa aaataaaatt tatttctttt ttgaatgata 3540 ataccgtgtc cagcgttgtc tgtctgtaat agttctatga attctttcca tggcttggta 3600 cactttagct tgaatgttaa ggtacatggg gatgagtccc tcgtcggggc tcaagtacag 3660 ccagtgcata caattgtcaa tgggctcagt attgtataca atccaatcaa agttagtatt 3720 gaggcggtgg tagttaaaaa tatccttaag caggagggaa atagcagtgg gttgtccctt 3780 agtgtagatg ttaataaacc tattaagctg ggaaggatgc accttaggac tgatgatatg 3840 catctttgct tggattttta agttggcaat gtttcctccc tggtctcgtc tgggattcat 3900 gttgtgcaag accactataa cagagtagcc tgtgcacttg ggaaacctgt ctagcagttt 3960 agagggaaag gcgtgaaaga acttggctat gcccctattg ccccctaagt cttccatgca 4020 ctcatccatt acaatagcta ttggcccttt agaagcggcc ctcgcaaaaa cattggtagg 4080 gttggtaaca tcatagttgg cctcattggt gagttctgca taagacattt gtacaaactt 4140 gggcatgatg ctgccactct gaggaaccac cgtgttttcg gggcctgcaa gaaagtttcc 4200 ctcacatatt tgggtttccc atgctgcaat ttcctgggga ggaatcatgt ccacctgggg 4260 ggtgataaaa aagactgttt ctggggcggg aactataagc tgggcagaca tgaggttcct 4320 aagaagctgg gacttgccac accctgtggg cccatacact gcagcaataa agggttgctt 4380 gtagtagtta agggatgcac agcttccatc tttattaagg tacttattga ccttcatgca 4440 acactgggtt atgctatctt gaatatctag taggtcattt aacagactgt ctcccccccat 4500 agaaagcagt tcctcaaaag aggcaaagtt tttgaggggc tttagtccct ctgcatatgg 4560 catgtttgtt aggctttgct ggagagtttg caacctgtcc cagaggcttt ttaagtcatc 4620 cactgcagct ccatccaggt agtttctgtg tttcttgggt ttggatgctt gcggctgtat 4680 ggaatgagac gatgcatgtc tagagcatgc agggttttgt ccttccatgg cctcagcttc 4740 cttgttaggg tggtttctgt cactgtgaaa ggttgcacgt gggcctgcac gctggtgagg 4800 gtcctgcgca ggctcatcct gctggttttg aagacgtccg agccctgctg catgtcggca 4860 tagtagcaag cttttagcac atcataagag agggtggaag tggcatgacc tttggccctg 4920 agcttgccct tgcccacgtg atggcactta gggcaggtgg tgtccttgag agcatacaac 4980 ttgggtgcca ggtacactga ttctgagctg taagcatccg atccacactt ctcacactgt 5040 gtttcacact ctactagcca agtgagctct gggttgctgg gatcaaagac tagcttgcct 5100 ccattttttt tgagtcgttt cttacctttt tcttccatca atttgtaacc ttccatggtg 5160 gtgaagaggc tgtcagtgtc tccgtagaca gatttcagga ccctgttttc aagtggggtt 5220 cccgcgtcgt tttcgtagag aaactgagac cactctgaga caaatgcccg ggtccatgct 5280 aggacgaagg aagcaatttg tgagggatat ctgttattgg ctattagggg ggtggacttt 5340 tctaaagtat gtaaacaaaa gtcatcatcc tcagcatcaa taaacatgat tggcttataa 5400 tggtatgacg tcatgctaaa attatgcata aaaggcgctt cggcgagggg gtctttagtt 5460 ggatcttctt cgctgcaaga aggcggtgcc aactggtgaa cgtcagaatt gacaggtggg 5520 tacgctacga caaattctgg catgatttct gcacacaaat tgtctgtttc tatgtatgag 5580 gatgatttga ctacataggc accagaagca atttctttag cgatgtttga ctctatttga 5640 tctgaaaaaa ctgttttttt gttgtctagc ttggtagcaa aagacccata gagggcattt 5700 gacagcagct tagcaatgct gcgcatggtt tgatttttgc ttttgtctgc ttcctctttg 5760 gcactaatgt tgagctgcac atactctctg gcaagacatt tccactctgg aaacacggtg 5820 gttctggtat cagggatgag ggtaaccttc cacccgcggt tgtgcagggt aataacatct 5880 atagaggtcg caatctcccc gcgcaggggc tcgttggtcc agcaaagcct gcctcccttg 5940 cgtgaacaaa atgggggcag cacgtccaga aactcttctg ccggtgggtc agcatcaatt 6000 gtgaagatgc ccggaagaag gtccttatca aagtagctaa tagttttgtg atttagcatt 6060 ttttgctcat aagcctttat ggccagcgcc cgctcatagg ggtttagcgg tgaaccggcg 6120 ggaaaagggt gtgtaagggc acttgcatac atgccacata tgtcatagac aaacactggc 6180 tcttgcaaca ctccaatgta agtggggtag cacctgcctc ctctgatgct aagccttaca 6240 tactcataca tttcattgga gggtgccatg aggatggtac ttaggtggtg tgtgttaggc 6300 ttttctgcct tatacaagat ttgtttaaaa atggcatggg agttggagct gatggttgga 6360 cgctggaaga cgttaaagga ggtttcaggt aagtttacag agtttttaat aaagatttgg 6420 taggattcta tgagtttgtt taccaactgg gctgttactt gaacgtcgag tgcgcagtaa 6480 tcaagggttt gctgaattat gtcgtaagcg cccttttttt cttttatcca aaggtctttg 6540 ttaaaggagt actcctcctg atccttccag tactttaagt ccggaaagcc atcggcgtct 6600 tgttggtaag agccaagcat gtaaaactca ttcacagcct ggtagggaca gcagcctttg 6660 gaaaccgtta ggctgtatgc tttggcagca tttttgaggc ttgtgtgtgt taaagagaaa 6720 gtgtcacgca ccataaattt tatgtactgg cttttcaagt cagacgccag tacttggccc 6780 tgctcccagt gcttgtaact cttggcaggc acgtagctgg ggttaggcag agagtaagta 6840 atatcattga acagcaacct tccagctcgc ggcatgaagt tgcggcaaat ttcaaacaca 6900 ggtggtattt ctgcccgcgt gctaaccacc tgagctgcta gcaatatttc atcaaaccct 6960 gtgatgttgt gtcccactgc atacacctca ataaatcttg ggtttccttt gagcttaaat 7020 gtttttaact gatctggagt aagctcttca gggctttcta gtcccaaagt agcggcttct 7080 tcttgaagct ggggattttg gcaaatcacg tgggaccaga ggctggcggc aatatgctgc 7140 tgcaaggtgt ccctgaactt tttgaattta actcctatgg ctcttttttc tggagtgatg 7200 caatagtaca gggtgtcctg ttctttacca ttccaaattt cccagttttg ttctgttgca 7260 agatttttgg caacattgac caggtggttg tcccccagca gttggaaaac tagcataaat 7320 ggcacaagtt gctttccaaa cttgccgtgc cacgtgtacg tttccacatc atatacaata 7380 aacagcctct ctgtgttggg cggggcccca atgggggtga aagatatttt ttccccaccag 7440 tctgcgctgt gtgcggccac gtgatggaag taaaagtccc gtcttctgag ggcgcaggtg 7500 tgagtaactt taaaaaaaga gccgcagaac tcacactttt gcatttgaga gatgtctttt 7560 attaggaaga ctttaccttg cttaactaaa aacttgagtg gaaagctgag caacggaacc 7620 ttcgcttgaa actccaccct cctcactgtt cctcgttggt agtagcaaat tagactgtag 7680 ggtgccgcgg cctcctctat gagggcgggc aggttgtggg cagtgatggg cctgtttaag 7740 catgcaggta ggtcatagag gttagtgttg caaaggttag taagcgcgct ggccaagttg 7800 gagtgatact taatttccac acactgtcca ttatccaaca cggcggaaat gcttagtgtg 7860 gcgcgctgga ccaccacggc gcccttgtac ttgcgcttgg gtgggattac tatggtggaa 7920 gttgcacctg ctcgtggtga gctggcaaag gcctgtgctg catcctgagc tgggaggcta 7980 gggccactac tctcctgttg attgattgga tttggcggtt ctgtgaaaag accactggtc 8040 ccgtgacctt gaacctaaaa gagagttcca tagagtcaac ctctgtatca ttaatggcta 8100 cttgtctgag aatttcgctc acgtccccgc tgttttcatg ataagcaata tctgccataa 8160 actgctccat ttcctcttcc tctagctccc ctagcccagc cctttccacg gtggcagcca 8220 ggtctcccga cacgcgggcc ataacctggc taaaggcatt ttctcccatt tcattccaca 8280 cccgactgta gaccacctgc ccctcgtggt ccctagcccg catgaccact tgagctagct 8340 ctaggtcaac ccagcgggag cagggtgggt acagcctaag gttgtggtgc aggtagttga 8400 gggtggtggc tacgtgctca gctacaaaaa aatacatgac ccaacggcgg atggtgagct 8460 cattaatgtt gcccagcatt tccaggcgct ggatgacctc ataaaagtca actgcaaagt 8520 taaaaaactg ttcattgcga gcagacactg tgagttcctc ttgcaggagg cggatagctt 8580 ctactatagt gtctcggact tcttcttcaa aggaacggcg aggaggggag gcaggaggag 8640 gagaggcctc tgcttcgggc tcagacagct cctcagacac cggaggcggc gggggagccc 8700 tgcgtctgcg cctgcgcact ggaagcctgt ctataaacct ttctatcacc tctccccgcc 8760 tgcgcctcat ttcctctgta actgcacgtc cattttccct gggcctaagc tcgaaggctc 8820 ccccagagag cggggtgcag ggcgcgggtc tgggcaggct cagggcgcta attatgcttt 8880 ttgttacaga ttggctgggt aaagactgaa ttgtttgaaa ctgtgacagc tccggatcag 8940 tgaatttttc cacaaaggca tcaagccaca aacaatcaca aggtaagctt agcatggtgt 9000 gcgggttgcg ggggctaact aaaaagttga aaaaagctgt cttaagctta cggatggtgg 9060 ttagaataac taggtctttg ttgcccgcct gctggatgcg catgcggtct gccagtcccc 9120 atgcttcgtt ttgacactgg gacaggtttt tgtaatggtc ttgcaagagc ccctctacgg 9180 gcacctgcct gtcgcctgcc atgttggtgg tgccatagcc cctaagtggc ctaataaggg 9240 ccaaatcggc cactacgcgc tctgccagga ccgcttgctg aacctgcgcc agggtctctg 9300 caaagttttc aaagtcaatg aacctgtggt aagctccagt gttaatggtg tatgagcagt 9360 tggccatcac cgtccagttt agtttttgct gcaagggcag gtgtatctct gtgtacttta 9420 gcctgctgta ggacctggtg tcaaagacgt agtcattgca cgttctcact aggtactggt 9480 agcccaccaa aaggtggggc ggggggtacc aaaacagtgg ccagccctcg gtagcgggct 9540 ctcgagggct gaggttcata agcatgaggc ggtggtattg gtatatgtac ttggacatcc 9600 atgccagccc tacagaggtg gtggaagctc tggcgtactc tcgaacccgg ttaaaaatgt 9660 ttctgagagg cctgaacacc tcaacggtgt atggggtttg tccggttagg cgggcgcagt 9720 ccagcgcgtt ctgcaaaaca aacaaggagt cgatggcact ttcattcgct ccattgcaga 9780 tgcatccggt actgcgacag atgaagcccc tgcccgcctc tgccagcaag ctagaagcag 9840 agagcgaggg tttggcccgc ttacaagggg gtgctgctcc agagttgcac ccccgggtgc 9900 agatgaagca agaggcggcg gaggcttaca tccccgctag caatgtgttt agggataatg 9960 aaggagaaga agcagagggc ttgcggcact taaagtatga gtcggggagg ctgttgcgta 10020 atgaccacac ttccaagcgg gtgctgggcg agcgagactt tgaaaaggac ccccagaatg 10080 gcattagcgc ggccgaggca cacctaaagt ctgcagactt agtcacggcc tatgagcata 10140 ctgtgaaggc agaggttaat tttcagacca cctttaacaa caatgtgcgt accctcatcg 10200 ccagggaaga ggtggtcatt ggcctgatgc acttgtggga ctttgtggag gcttttctag 10260 aaaacccagt gagcaaggcc ctcactgccc agctctttct tatagtgcaa cactgcaggg 10320 acgagggggt gctgcgggaa agcttgctaa acattgccga gcctgagagc cggtggctgg 10380 tggacctgtt aaacctgtta cagactattg tggtgcaaga gcgaggcttg gcagtcggtg 10440 aaaaggtagc ggctataaac tactctgtga ttactctaag taaacactat gcaagaaaga 10500 tattcaactc tgtgtttgtg cccatcgaca aggaagccaa gattaacaca ttttacatgc 10560 gcactgttgt taagctgctg gtgctgagtg atgaccttgg catgtaccgc aatgagcgga 10620 ttgagagggc agtgtctggt gccaggcagc gggaattaaa tgacagggaa cttatgcacc 10680 gactgcggca agccttggct tctaacggac tgccggagct agagggcgaa gacagtgcta 10740 atattagatc caaagaggag tggggggtcg gagcgggcgg aggcgttgcg agcgcgcgct 10800 acccccacct gctagattat gaggaagagg agaaccctga tggctccgtc tcatttcaac 10860 agcatgagcg cggcgctcaa ccccatgaaa atggcggcca tgcagagtca gcctacagcc 10920 gacgacagtt gggccgcttc tattagtcgc attatgtccc taacggcggg ggatcggcac 10980 aagttttcat cccagccttt tgctaacagg ctggatgcta tattagaggc cgtggtgcct 11040 tcccgcaaag accctaccca tgaaaaagtg ctaactatag taaatgccct aatagaaaat 11100 ggtgccatta ggcgggacga gggggcgggg gtgtatgacg ccttgttgca ccgcgtgtcc 11160 aagtacaaca gcttgaatgc ccaaagcaac ctagagaggt tggctggaga cgtgagggag 11220 gcggtagcgc agcaagtgcg catcgccacg ggcaacctgg ggtccctaac cgcacttaat 11280 ggctttttgg cacgcctgcc cgccaatgtg gagcggggcc aggagaacta cactggcttt 11340 gtgtcagccc ttaagctgct ggtaagcgag gtgcccagta cagaggtgta ccagtctggg 11400 cctcactact ttttgcaaag cagtcgcaac ggcacccaga cagttaacct taccaatgcc 11460 tttgagaacc taaagcccct ttggggagtt aaggccccca ctatggaaag gctcagcatc 11520 tcagccctgc tcacccccaa cactagactg ctgcttttgc tagtgtctcc ctttacagac 11580 agtgttagca ttagcaggga ctcctacttg ggttacctgc ttaccctgta cagggaagcc 11640 ttgggccgca atcatttgga tgagcgtact cttgaagagg tcacagaggt cagcagagcc 11700 atgggcagtg agaacataaa caacttgcaa gccaccctta actttttgct aaccaataga 11760 caaaaaagaa ttcccaaaga ctactcccta accccagagg aggagaggat agtgaggttt 11820 gtacagcagg ccgtgagctt gcgcatgatg caggaaaact tgagccccac tgaagctttg 11880 gacgtaaccg cggccaacat ggagcccagc ttctatgcca gcaacaggga ttttattaat 11940 aagcttatgg actactttca ccgcgctgcg gctatggctc cagactactt tttgggggca 12000 gtgatgaatc cccgctggct cccccctgag ggcttcttta ctggggtttt tgactttcct 12060 gaaagggaca gctacacctg ggatggactg gatagctctt tggagctgac gcgccaggat 12120 gccatgcggt ttctggaaga caagtttatg gacgatgacc agaggacaga gtctcgcagc 12180 ctcagcagag ttaccacccc agctagctct aggcggtcct cggtgaccat gccacctggg 12240 ggctttatgg gcatgataaa taacaacaaa aacgataaca gcctcaggga gatagatggg 12300 ttagcagaca agctggctcg gtggaaaaca tacaagcgtg agtcagagga ggctcgtgtg 12360 tccctgcctg cggtagtaag gcctaagcag tacaggcccc gctcccctat ctccagcgat 12420 gacagtgatg atgggatgag caggccagac ccgtttctca agtttgaggg gagtggaaac 12480 ccctttgccc acctacgccc caagctgggg cgtggcctgt aaagtgaata aaatactcac 12540 cagagacatg acgctagcgt ttttttatag caatggagtt ttcgtcgtct cctcccccgt 12600 cttatgaaac agtgatggca caagtgcctt cgatcctggc gccgctggta cccccgcggt 12660 acaaaggggc tacagaagga agaaacagta tccgttattc ccagctgccg cctctgtttg 12720 acactacaaa gctgtacctc attgacaaca agtcttcaga cattcaggcc ctcaattacc 12780 aaaatgacca cagcaacttt ttaaccactg tggtgcaaaa tgctaattac acgcccatgg 12840 aggctagcac ccagtccatc cagctggacg agcggtcgcg ctggggtggg gaatttaagt 12900 ccattttaca catgaacatg cctaatgtga cagagtacat gtttagcaat agctttaagg 12960 cataccttcc cgccacagca gacaactttg gcaaggtgct cacctatgag tggtacaccc 13020 taaccattcc cgagggtaac tattctgagg ttatgctttt ggacctgctg aacaacgcgg 13080 tggtggagaa ctatctggcc cacgggcgcc agcataatgt tagggaggag gacatgggcc 13140 tcaagtttga caccagaaac tttcacctgg gctttgatcc cgagaccaag ctggtcatgc 13200 cgggcttcta cactaacgag gccttccacc ctgacatagt gctgagtcca ggctgtgcgg 13260 tggactttac ccacagcagg ctcaacaact ttttaggcat tagaaaaagg ctaccatacc 13320 aggagggatt tgtgattacc tgggaggacc tgcagggtgg caacattcca gccctgttag 13380 acctggaaaa ttacaaccca gacattcccg gcgccgatat cacaccactg atgtatgact 13440 ccaagggaag gccttaccat gttggtgagg atcccagcgc gggcagtacc ttcacctggt 13500 accgcagctg gtttgtggcc tataactacg gtcctgccga tggcatcaag agcaaaactg 13560 tactggtggc ccctgacatt acttgcggag tggagcaaat ttactggagc ctgccagata 13620 tggcggtgga cccagtaact tttacctcta gccacaatcc cagcagctac ccagtggtgg 13680 gcacggagct gctaccatta ctgccaagaa gcttttacaa tggatcttca gtgtacagcc 13740 agctgttgca agaaagcacc agtcaaactc aagtgtttaa ccgctttccg gagaatgcaa 13800 ttcttaaaag gccccctgct cccaccatta tcagcatcag cgagaacgtg cccgccctca 13860 gtaaccatgg caccctgccc ctaaaaaaca acattcccgg ggtgcagcgg gtgaccatta 13920 cggatgcaag aagaagagtg tgtccctacg tgtacaaaag cttgggggtc gttgtgcctc 13980 gcgtgctgtc cagtaaaacc ttctagtgac aaactttttc ttttcattag caaagccatg 14040 gccatcttaa tatcaccgac taacaacacc ggctggggcc taggcactca caagctcttt 14100 ggaggagcca aacaaaaatc agatcagcat cccgtttatg tccaagctca ctaccgggcc 14160 tcgtggggca gcaaagggcg ccgccgccgt caaggccgag cccgcggggc ccctcttgac 14220 cctaaaactg aagcagaaat ggttgccacc atcgacgagg tggcgcgcaa cggtcccccg 14280 gcggcgcgct tggtgctgga ggctgcccgg agagtggggg cttacaacct ccgtcgagcg 14340 cgcaagctaa cccctgcggg gcgtgccatg atggccatgc gcgcgcgtca gatggtaaag 14400 caggctaaga aaaggaagcg cagggttcgc tttagacaat aaagtctgtc ctcacataca 14460 cacatacccg agtgatggcc tattttttta ctgaagcctc atcgccatgg cagcaatcag 14520 tcgcgccatc aagcaggaac tgctggaaga cctcaaacct gaaatctatc tgccacccaa 14580 gtccaccaaa cggagaacta aagttaaaac agaggaaaaa gttgatgtga aaaccctagt 14640 caaggccaag agcagaaagc gcagggctgc caaggatgag ctggaagagg atgtagagtt 14700 tgttaggcgc tttgctcccc gacgtcccta ccagtggcgg ggccggcgcg tgcgcgctct 14760 gcctaggccc ggggtgcccg tggtgtttac tcccggccag aggtcgggca ccgcctcaaa 14820 aaggtcttat gacgaagtgt acgcagatga ggatgtactg gaccaggcgg gaaacatgat 14880 taatgagttt gcttatggca agcgagtcag ggtgcttacc cacaaaaacc ccactccctc 14940 acaagtgccc atcacccccc aggagcccgt ggcgcgtcct ggcgaggcac acctgctacc 15000 cacagtgcaa gtgctggctc cgcgaggctc taggcgggaa accatgttgc ccgtaaccaa 15060 gtccgagggg ggagacgtga aagtggaaaa caagggcttt gagcaaatca cccccgggct 15120 gggggtgcag actgtagaca tcaaagtgcc cgtgaagcgc aaaggcgatg cagaggatga 15180 aataataaag agggttaaga tggagcttga accatatgag accactatga aaatggagta 15240 ctctgaggag ccccaggttg aggcttttga cacagggata gagcccagct ctttttttga 15300 ggtgcgctct caagcgcgtc ccatcgctgt ggctaggaag cgccgcaccg cagctgccag 15360 cgcccccgct gtagaggtga tggaggttca gcaaagcaac cctgccaccc cagctaccgc 15420 tgccgccaga accgcgaccg cgctgggccc gcgtttggct cgtcggcctt caagatgggg 15480 gcccgccaat gccatctttc cagattacaa ataccacccc agcatcacgg ctaggaaaat 15540 aaggggcccc aagcctacag gcaagatcag tcgctggggg cctgccaaca gcatcctgcc 15600 tgaggtgcgg ctgcacccca gcatggtgtc tgcggtgacc agagctgccc ctcgcagagt 15660 caccaaaacc cggagacgcc ggcgtgccag aacccgccag gcttttgtgc tgcccgcgcg 15720 caccaaaacg ggcgcgctgc tttcccagaa cgtgcgctac cacccttcca tctccttgct 15780 ccgccgagcc taatctgtat gttttgcttt gcgcaaggat ggccggcagg aatgtcaccc 15840 ttcgtctgag agtcccagtg cgcaccaaaa ttacaggcgc aggtcgtcgg cgcggacggc 15900 gcccacgcgg catacgctgc gggcgcatga gaggaggctt tttgccagcc ctcatccctt 15960 tgatagctgc cgccattggg gcagtaccag gaatagcttc ggtagcgctc caagctgccc 16020 ggcactagct ttttacagat tgactcactc ccatttttct tgtgccagaa gaagacaaaa 16080 tggatgctgt caatttttcc atcttggcac cacgctacgg ggcccaccct atgatgagtg 16140 gttggtctgg catcggcacc agcggcatga acggcggggc ctttaactgg gggggcattt 16200 ggagcggcat caaaaacttt ggcagtaatg taaaaagctg gggcagcaag gcttggaaca 16260 gccaaacggg caagctgctc aggcaaaagc ttaatgacac caaagttaga gaaaagctgg 16320 tggagggcat ttccaccggg gtgcacgggg cacttgatat cgccaaccag gagttagcaa 16380 agcaaataga gcggcgtttg gagcgtcagc agcccctaga gccggaggtg gaggtagagg 16440 aggaagtagt ggacataaaa cccgaagcgc aggccccgct ggtggtacaa atacccaaaa 16500 agcgcccccg cgacgaagac ttgctcataa ctgcggacga gcctccctcc tatgaagagt 16560 ccatcaaaac catggcaccc ctgatgccca tgacccgccc gcacccttcc atggccaagc 16620 ctgttttggt agaccgtccc accaccttgg agctaaagcc ctctgaccag ccgccagcct 16680 actccccccc ggcgcccagc gccgtcaggg tcacagtgcc ctctaacatt cccgtggtca 16740 ctcctgccag aagcagaggc tggcaaggaa ctctggccaa cattgtagga gtgggtctaa 16800 gtaacgtgaa gcgccgtaga tgcttctaac tgtgattaaa tatactcaag tgacttgaag 16860 tttgccgacc ctgtcttttt ttgaccgcgc cagctgagaa gatggcaacc ccgtcgatgc 16920 tgccacaatg gtcttacatg cacattgctg gccaggacgc cgccgaatac ttgtctcccg 16980 ccctggttca gtttgcccaa gcaaccagtt cttactttaa gttggacaac aagttcagaa 17040 accccactgt ggccccccacc cacgatgtga ccactgagag gtcgcagcgc ttgcagctgc 17100 gctttgtgcc agtcatgcaa gaggatggcc agtacactta caaaacccgc ttccagcttg 17160 cggtgggaga caacagggtg ctggacatgg ccagtactta cttcgatatc aggggtaccc 17220 tagacagagg cccctccttc aagccttaca gcggcaccgc ctacaatgcc ctcgccccca 17280 aggccggggc taacaactgt ctttttaatg gacagggtgc caatattaac actttagccc 17340 aggtgccctc tgcaggtgcc ataactgtga atggccaagc tgctgtcaca aacaatacct 17400 accagccaga gccccagctg ggccctgaaa gctgggtcga tggcagccta gcagagctgg 17460 gagatgcgtc tggccgtgcc cttaaggctt caaccccgcg catgccttgc tatggttcct 17520 atgctccccc caccaacgaa aatggaggtc aagcaactgg tccagtggaa tccagatttt 17580 ataaggtgac caccaacaat aacaatgaag cagatgccat gctatacact gaagatgtga 17640 acctgcaggc cccagacacc cacctggtgc accaagtacc agagggtcag gttacagggg 17700 tacaagggct gggccagcag gctgcaccca acaggccaaa ctacataggc ttcagggaca 17760 acttcatagg cctcatgtac tacaatagta atggaaacct aggggtgctg gcgggtcagt 17820 catcccagct caatgccgtg gtggacttgc aagacagaaa cacagagctc tcttaccagt 17880 tgctgctgga tgccctcaca gacaggtccc gctacttttc catgtggaac caggctgtag 17940 atagctatga ccaggatgtt aggattattg acaaccatgg cgtggaagat gatatgccca 18000 actattgcta cccactgagc ggcatggggc ccctaacaaa catgaccacc atgaaggtta 18060 acaaccagaa ctttcaggca gaaaatgcca atgtgggacc cattcaaaaa attggttttg 18120 gaaatgttga ggctatggaa atcaacctca atgccaacct ctttaaaagc ttcctttact 18180 ccaatgtggc cttatacctg cctgatgcct ttaaatacac acctgaaaac attgtggccc 18240 ctgccaatgc gaatacctat gcttacatga atgttagatt acccgccgcc aaccttatag 18300 ataccttcgt aaatattggc gccagatggt caccagatgt aatggacact gttaatcctt 18360 tcaaccacca cagaaatgca ggactccgct accgttcaca actgcttggc aatggccgct 18420 attgctcgtt ccatattcag gtccctcaaa aattttttgc aatcaaaaat ctcctcctac 18480 tgcctggaac gtacacgtac gagtggtctt tcagaaagga tgtaaacatg atccttcaaa 18540 gcagcttggg caatgacctc cgagtggatg gggccaccat caacattcag agcatcaacc 18600 tatatgcaag cttttttcca atggcacaca acactgcctc cactctggaa gccatgctgc 18660 gcaacgatgt aaatgaccag tcctttgcag actacctgtc ttctgccaac atgctttatc 18720 ccatccctgc caacactact aacctgccaa tttccattcc cgctagaaac tgggcgggat 18780 ttagagggtg gagctttacc agaattaagc aacgagaaac tcctgccctg ggctcgcctt 18840 atgatcccta cttcacttac tcgggcagta ttccatatct ggatgcaact ttttacctca 18900 gccacacctt tagaagagtt tccatcatgt ttgactcttc agtgtcttgg cctggcaatg 18960 acaggctgct tacccccaat gagtttgaga ttaaaaggta tgtagacggt gaaggttaca 19020 atgtggccca gtccaacatg acaaaagact ggtttatggt tcaaatgcta gcccactaca 19080 acatcggcta ccaaggctac cacctgccag aaagctacaa ggacagaatg tactccttcc 19140 taagaaactt tgagcccatg tgcagacagt tggtggacgt ggccaactat gctgcgtacc 19200 agccagttac cgtgggccac cagcataaca attctggtta tgctagcgcc ctttcggcct 19260 ttaacccacg tgagggacac ccatacccag caaactggcc ttacccactc attggagcca 19320 atgcagtacc cactgtcacc cagaaaaagt ttctctgcga caggtccctg tggcgcatcc 19380 cattctcctc caactttatg tctatgggaa cccttactga cctgggccaa aacctgctgt 19440 actctaactc cgcccacgcc cttgacatga cttttgaggt tgatgccatg aatgagccca 19500 ctctgttgta cgttttgttt gaagtgttcg acgtggcacg cgtccatcag ccccaccggg 19560 gggtgattga ggtagtgtac ctcagaactc ccttctccgc cggcaacgcc accacctaag 1920 ctaatatggc ggaaggaggg tcctcagaag aagagctgcg agccatagta cataacttag 1980 gcgtgtctcc atttttcttg ggcacctttg acaagaggtt tccgggtttc atctcttcac 197440 aaagaatggc ctgcgccata gtaaacaccg caggccgaga aacggggggc gtgcactggc 1800 tggccatggc ctggaacccc cgctcaaaaa cgttttacat gtttgacccc tttggatttt 19860 cagacactaa gctcaagcaa gtgtacagct ttgagtatga gggcctactc aggcgcagcg 19920 ccatagcctc cagcccggat aggtgtgtta ctttggccaa aagcaatgaa accattcagg 19980 gtcccaatag cgccgcctgt ggactgtttt gttgtatgtt tttgcacgcc tttgtcaact 20040 ggccggacga cccctttgac cacaatccca ccatgggacc cctcaagagc gtgcctaact 20100 acaaactcaa tgatcccaca gtgcagtatg tgctctgggg aaaccaagaa aagctgtata 20160 agtttttgga aaaacactct gcttactttc gcacccacgc cgctgcaatt aaagccagga 20220 ctgcttttaa taaattgaaa caataaaccg tttattcgaa aaatataaat ggtttacagt 20280 gtgattatta aaaaaggaca aggtcatcgt catgttggcc ctggggcaaa atggtatttt 20340 ggtacctgtg ttcatcggac cagtggaact ctgggaattt aatggggggc ctttcgccca 20400 ctgtggacgt ccatatttgc ttagcaattt ggacacatga caccaggtcc acgctggaaa 20460 tcttaaaatc acaatttttt tggggggcgg ccttgctgtt gcggtacact ggattgcagc 20520 attgaaacac tagcatggcg gggttattaa gggtggctag cattttttgg tcatccactt 20580 cacttttgtc aatgtgacta gcgcttgcaa gagagaatgg ggtaatcttg caagtttgtc 20640 tgcccagcaa aggaaggtta ctgttccaat tgcactcaca cttgagtggc attagcaagt 20700 gactctcagc atttggcata gaggggtagc aagccttaag aaaagccatg atctgctgaa 20760 aggccataat agctttgggt ccatcggtat aaaacatacc gcaagaggcc ccgctaaagt 20820 tgcccccgct catattaaca tccatgctgc agcagaaagc atcctcattc ttaaactgta 20880 ctacattcct tccccaccgg ttagaaacaa ttttggtttt ttctgggttt tccttaaggg 20940 cccgctgggc attttcactg ttaacatcca tttctactag ctgctctttt tggatcatgg 21000 gcgacccgtg cagacacata aggctgcttt ggcatccatg ttgccacacc acacacccgc 21060 tggggttaac cccaggggca agctctgctg ctttgattac aaaatcaagc agcaggcgcc 21120 ctgccacgtg caaaaaagtt ttctggttgg taaaggtgta agtaacgtgc gttttagagg 21180 tggtaatgta agcctggact gcttttttaa agcactccag tgtgccctcg tctggaagca 21240 aggttagctt gctgtggtct acttggtacg cggtgagaac cttaagggca agctccatgg 21300 cggactgcca cttgtgctca ctagctttaa aaaacttaga agccactggc cccgagggcg 21360 cgttgggctt gctccaaacg gacttttgct tgggtgggat aaataccaca tttccctcct 21420 tgtcagtgtc caccgtttgg ccacaaaagg tctgcactac aatgctcccg ggatcccccc 21480 gcgccttgag ggtggcggtg gcttgacggg ctatttcgac catggcttgg tggccccttg 21540 gctcctcaac ggctttccgc tggcgcttct ttgggggcgc cgtaggcggt gccacaggaa 21600 cctcttcatc gctggagctt tcagagattg caacaacctt cttgtggctc atctttttcc 21660 tagatgtcag aagagcccgt cagtgggact accgtggaga caaaggagga cattcacaca 21720 cctccaaact cccctgtcct cgagacaatc tctctaagcc ccgaacccga ggctgaggcc 21780 tgcccaaata cagacaaata cctaagcacc aatttgcttt gcaaacacct gcaacgtcaa 21840 agcgcaattg ttctggacag tatcaaagat catctacaag tgcccaccag tgtatcagag 21900 ctaagctgcg cctacgaacg aagcttattc tccccaaaca ccccaccaag gcaacaaagc 21960 aatggaacgt gtgaagcgaa tcctaaactt aacttttacc ctactttttt ggtgcctgaa 22020 actttggcaa cttatcacat cttttttgtt aatcaaaaga taccagtatc ttgcagagct 22080 aacagagcta aagcagacag agctcttgct ttgcaagagg gagattgctt acctgactat 22140 gaaacaatgg acacagtcag ccgggtgttt gagggtttag gcggtgaggt ggttgcggat 22200 cacgcgctgc aaaataatga ctctgtatta gtggagctta aggaagacaa cccccgtttg 22260 gctgtggtca aaagaaacct cagcgtgtcc cactttgcct accccgccgt gcacctgcct 22320 ccaaaagtca taaccactgt catggacaac ctgctagtta aaaggtccac ccccagcgcc 22380 gacgtctcag agctagatcc cgaggggggc caagaggtgg tgtctgacac cgagctaagt 22440 aagtggctaa acacctctga ccccgaggcc ttagagaagc agcgcaagac agtaatggga 22500 agcgtgctgg tcactgtagt gcttgagtgc atgcaaaggc ttttcacctc agaagaaatg 22560 gtgaaaaaaa ttggggaggc cctccactac acctttaggc acggctacgt gtctctggct 22620 tgcaagattt ctaatgtaga gctgaccaac gtggtcacct atatgggcat cctgcatgaa 22680 aaccgcctgg gtcaaaccac cctgcatcac acagtgcagg gcgaggcccg tcgagactac 22740 attagggact ccatcttcct catcctaatc catgcctggc aaaccgcgat gggcatctgg 22800 cagcagtgcc tcgaagagga aaaccttaaa gaactagcca aactgctgca aaggattaaa 22860 aagccattgt acactgaaac ctcccagcgc cttatgggca agctcctggc aaacgtagtc 22920 ttccccaccca aactgctaca aaccttccac aagggccttc ctgatattgt tagccaaagc 22980 atgatgcaaa actttaggtc ttttattttg gaaaggtccg gcatcctgcc ctccatgacc 23040 tgcgcgctgc ccacagattt tgtaccaatt cactttaaag agtgcccccc cacgatgtgg 23100 ccctacacct acttgctcaa attagcaaac ttctttatgt atcacaatga cctttgctat 23160 gatgtgggag gcgagggcct gctagagcac tactgccggt gcaacctctg caccccccac 23220 cgttgcctag ccaccaaccc cgccatgctt aatgaaactc agctaatagg tacctttgat 23280 attcggggtc ccggcggaga aaacggagca gagtcttcct ctggccttaa gctcaccgct 23340 ggaatgtgga cttccgcgtt tctgcgaaaa tttgaaagct ctgattatca cgcccacaaa 23400 attcagtttt atgaaaacca atcaaagccc ccctcagtgg agcccactcc ctgcgtcatc 23460 acccaaagta gcattttggc ccaattgcat gacattaaaa aagcaaggga ggagtttctg 23520 ctaaaaagcg ggcagggtca gtatctagac cctcagactg gggagccgct caacgctgcg 23580 gatccttccg tagaaagcag ccatgagttc caaggagatg gaagacacag aaaccctaag 23640 cgtggaagaa atttccggca gcgaggagga cctagaaagc ctcccagagt ccatgccgga 23700 gcagagcgag atgccagagg aacgagctcc tagatgggac caaaaagaga agcctctggg 23760 taagcagccc cgcaactacc actcctggag agcgcataag tttaagattc ttagctgcct 23820 gggcgccagt gggaacaatg tggcctttac cagaagatac atgctctttc gcgagggagt 23880 taaccttccc aacaacatca ttcactacta taactctcgc taccgcagca aaacagaagc 23940 tcaagcgcag accgacccca ctaaagagcc caaagccagc cgccggtgag gacaaccacc 24000 agacaaggca gctgcgcaac cgcatttttc caacactgta cgccatcttc caacaaagca 24060 gaggctcccc cacggcatta aaaataaaaa acagatccct cagatccctc ctaagaagct 24120 gtctctacca caagtcagag gagcagctgc tgcgcacgcg aaacgacgcc gaagctttgc 24180 tcaacaaata ctgtcaagga ctcgagtccg gcacagactg agcaatgtct aaagaaatac 24240 caacccctta tatgtggagc taccaaccgc aaacggggca cgccgccggc gcctcccagg 24300 actactccac ccaaatgaat tggtttagtg ctgggccatc aatgattagt caagtttatg 24360 gcattagaga cttgcgcaac aaagttttga taacccaggc agaaataacc aaaactccca 24420 gaacaataat ggatccgcca atttggcctg ctgccatgct tgttcaggaa gccgccccac 24480 ccaaaacggt cactctgccc agaaaccaca ccctagaaca ggctatgacc aactctgggg 24540 cacagctagc gggaggacga cagctgtgcc cctcccaaat aggtataaaa agcccagtgc 24600 tggctggcac gggcattcag cttagcgaag acatccccag cgcctcctgg atcaggcccg 24660 acggcatatt ccagctagga ggggggtccc gctcgtcctt cagcccaacg caagcattcc 24720 tcaccctgca acaggcatcc tcgacgccgc gcgcaggagg cgtgggcacc taccagtttg 24780 tgcgcgaatt tgtgccagag gtatacctta accctttttc aggaccaccg gacacctttc 24840 ctgaccagtt cattcctaac tacgacattg taaccaactc tgtcgatggc tatgactaag 24900 gagagcatgg accaggtgga ggtgaactgc ctgtgtgctc agcatggcca aacctgcacg 24960 cgccctcgct gctttgcaaa agagggttta tgtgctaact ggttttacaa cccagcactt 25020 gcctttgaag ggtttgatat tccagactct taccaagagg gacacggtgt ggacatagaa 25080 gttaagtgtt cccaccactc cagcaaactg tgtcacaatg gccatgatat gatctgctca 25140 tactctcgcc tgggatccca cattaacata agatgtattt gcaacaagcc gcggccccac 25200 atgagcctca ttgaggcagc ctgttctatg tataacctta actagataat attattaaac 25260 ttgttttaca gctaccacca taatgcgctt cagcttcttc atcgccgccg tccttttctg 25320 caccacaggg gctagcaatg acatgtgac ttgctgcgcc cacacacctt gcctcctaca 25380 cctagaagtg ggcttggggg ccaatgtcag ttggataaac tctgacacag gccaggcccc 25440 gctttgcctc tccaatggca tgtgcaacgc tacccagcaa ggcctgcagt tttctgcaaa 25500 cttttctgag gatggcctgt acatcgccct cattaaggag agcaactacg agggcgctga 25560 gcactactac cttgtttata tttatggaga ctgctaccaa actgcaaatg agtctgccca 25620 cgggcctatt tccagacccc tcaacgagat gcctcttcac agcgtaacca taaatgcttc 25680 cctcttctat cccgcctttc tggagctgcc cccacagtac agcaatgacc ttagcaatgt 25740 gcgctggtat aaagtagacc ccagcggctt ccaagcccaa aaaatctcta aagtcagaag 25800 cggaggcaga aaagagaacc tgcatcccaa ctgggccttg gttacctata ctggagacct 25860 tcttgtcttg catgtctcgc caaacaccct tggactgtgg ctggcagccg tgcagcatcg 25920 cggggggacgc actaatttca ttaccttcaa cataactgta cccaactggc aacaaaatct 25980 agtagccata tttaatcaac acgagccccc aaaaatgggc aataattatg aggacagttt 26040 tatggaatgg actctgttta aaaagctcaa aaaaggctta tttagagtaa cttgcagagc 26100 caagtcaata tttccagagt gcaccctcaa catcacccgc gacggaactt tcctgcttat 26160 tggggaaagc aagaagaccc cctatgtcat cctgctgccc ttttttgcaa accccaaaga 26220 agacactcca attttaatgg cccttagcca ctccgtgccc gttgccatac ctaacactgc 26280 aatgcctgta tatatttcca tcatgttttt tattgtagcc atgctagcca ccctcagcct 26340 tctaatggga ctaaacaaca aaatcaggtc catgtagctt gtcaaataaa cttacctaat 26400 ttttgctaag acgtctgggt cctgcgtttc tatgtccacc aaagtcccct cttcccagct 26460 ttggtacttc cacttgtgtg tgcgagccag cttgcggatg tgcttgaaag ataatgtggt 26520 ctctcccaac agcttcccgt tcaccagcac cagggccatg aagcggacac gaagagctct 26580 acctgcaaat tatgaccctg tgtatccata cgacgccccc gggtcttcca cacaaccccc 26640 attttttaat aacaagcaag gtctcactga gtcaccccca ggaaccctgg ctgtcaatgt 26700 ttcccctcca ctaacctttt ctacgttagg tgccattaaa ctttccacag gtccccggact 26760 caccctcaac gagggcaagt tacaagccag cttagggccc ggcctcatca caaataccga 26820 gggccaaatc actgttgaaa atgtcaacaa ggtcctgtct tttacctccc cattacataa 26880 aactgaaaac actgtatccc tagcgctagg agacgggtta gaagatgaaa atggcaccct 26940 taaagtaacc tttcctactc cccctccccc gctacaattc tcccctcccc tcacagaaac 27000 aggtggtact atttccttgc ccctgcaaga ctccatgcaa gtgacaaatg gaaaactggg 27060 cgttaagcct accacctacg cacctccctt gaaaaaaact gaccagcaag ttagcctcca 27120 agtaggctcg ggtctcaccg tgattaacga acagttgcaa gctgtccagc ctcccgcaac 27180 cacctacaac gagcctcttt ccaaaactga caattctgtt tctctgcaag taggtgccgg 27240 ccttgccgtg cagagcggag ctttggtggc aacccctccc ccgcctctca cctttacatc 27300 acccctagaa aaaaatgaaa acacagtgtc gctacaagta ggcgcgggct tgtctgtaca 27360 aaacaacgcc ctagtagcca cacctccccc acccttaacc tttgcctatc ccttagtaaa 27420 aaatgacaac catgtagctc taagtgccgg aagtggttta agaatatctg gaggcagcct 27480 cacggtggcc actggacctg gcctttccca tcaaaatgga acaatagggg ctgtggtagg 27540 tgcaggcctc aagtttgaaa acaatgccat tcttgcaaaa ctaggcaacg gtctaaccat 27600 tagagatggt gctattgaag caacccaacc cccagctgcc cccataactc tgtggactgg 27660 gcctgaccct agcattaatg gctttattaa tgacactcca gtaattaggt gctttatatg 27720 cctaaccaga gacagcaact tagtcacagt aaatgctagc tttgtgggag agggggggta 27780 tcgaatagtc agccctaccc agtcacaatt tagcctaatt atggagtttg atcagtttgg 27840 gcagcttatg tccacaggaa acattaactc caccactact tggggagaaa agccctgggg 27900 caataatact gttcagccac gcccaagcca cacctggaaa ctgtgcatgc ctaatagaga 27960 agtttactcc actcccgccg ccaccatctc ccgttgtgga ctggacagca ttgcagtcga 28020 cggtgccccc agcagaagta tcgactgcat gctaattatt aacaaaccaa aaggcgttgc 28080 cacttacacc cttaccttta ggtttttaaa ctttaacaga ctaagcggag gtaccctgtt 28140 taaaactgat gtcttaacct ttacctatgt aggcgaaaat caataaaacc ataaaagaaa 28200 taagtttaaa agctttattt ttcatacacg cgagcggtaa ggctgccgcc ttcaggaaaa 28260 gttactttgt aaacagttct ttcacaacag cacaaaacat aggtattagt taacagttca 28320 tttgggctat aataatatac attttcttgg gtggcaaagc aagggtcggt aatctcaaca 28380 aaaccatcaa ctggaatgca agaatagtcc agcacggtgg gttcaatcta aaaatgaaga 28440 aacgcgttga ggttcactaa gcacaggttt cgaatctgtc ggcagcgtcc atacatcata 28500 gcttgtctca aagcagattg tcttctttcc tctgccttgg aagtagtttg gtgaagcact 28560 acaggtgtct tttcaacctc tttcagcacc cgctctatta cagatctcac ccacacagca 28620 cagtttttaa gagaacaata gttttgaagg ctgcaagatt tacacttaag caccagccag 28680 taattataag tgcttttaag aactacccct agctcagggt taatgcacct tttaatggcc 28740 tccatgcagg ctttatggac agttctaaaa aaagacagtc taaaataaat gtagtgagtg 28800 tttctaaata taatactccc cacatagtta atctcatcag gcctgctaga atttacaaac 28860 tctcgatacc acatatactt tttattcata gccccaccct taacaaagtc ctcaatcact 28920 ttctgaacca catgcttgct agccatgcat tgtaaagaca agctgttaga acagtgacag 28980 tgtaattgcc acgtttgagc ctctgccagg cagcagtgct tagttactat caactcaata 29040 cccgcattgc atgtaaaccc cccaaatagc agtttttcat gcctgtgtag cacatcatcc 29100 cacaaaatag gaatttcata gcataaagca aagcaattac aatatttagg aactctcacc 29160 acagcagtca cgtggcatgt tgtctcagca gtgcagttgc cttccatcct acaattatga 29220 acaaaaacta aacacttcta acaaagatac agtgacaatc tcccttcctc taaaagcatt 29280 gtttacatta gggtgattat taacaacgtc agaaatttct ttaattaaag tgcctttaaa 29340 atgtgcaaga gcatcatcat actcaaaacc aagctgagag taaaaaacca ccttaaaagt 29400 aatcccaggc ttgtttttat caacagcctt aaacatgctt tcacaaaata tagaagcagt 29460 aacatcatca atggtgtcga agagaaactc cataggagac tccagcattg atccaagctc 29520 tctaacaaaa tcttcctcaa aatgaataat gccctttaca caaacgcggg gcagacgatg 29580 gtgggccatc gcgtcaacct gaaacacatt ttacagtaaa caaagctagc tccgcagtgg 29640 taaagtcatg cccatgggtg ataccaaaat ccttaaaaaa gctatctaag tagttggtca 29700 tcccctcagt taaaaagttt tgcagctggg tggtgcatgc cacatagtgc cagcttatag 29760 ctacaaagac ctccatcccc tccttaacag gcagctcttg cacacacgca gtaactatcc 29820 accgcttaag aaaagcttta agcccagcgc acataacagc tccaatgttt ttatccaagg 29880 agagcaaaat ttcagcaagc gcaggctcca cagtaatagt aaagcagagg catttcagac 29940 gaggctcact atctgcagtc gccatttatg aggtctgcaa taaaaaacaa ctcatcagca 30000 gctgaaaaag tgcactttga cctcattaag ccactgcata tgcaagtcct catctatgcc 30060 gcaacccaga ccctcaatcc agccccgaat gtacacttta ataagagatt caacctcttc 30120 ttttagtaaa gtgcacatgc tgtttggact agtatacaca atagaagtca caatgagggg 30180 cccgctgtga ctggaaagcc tgcgcacagc ccgaaggtta aaaatggact gtaacagcat 30240 tgaaaccccg cgacacaggt cagtctcgcg gtcttgatct cttattatag cgaccaaatg 30300 gtccttcaaa gtaatgttgc actcatagaa gtaggcagct ccggaagcca ttctgcaaaa 30360 taacaaaaca ccactaagca tagcaccgtc accaagcatg aaaacaggta aaaacaaaag 30420 caacacttac ttattcagca gtcacaagaa tgttgggctc ccaagtgaca gacaagccta 30480 atgcaaggtg ggcacagtcc tccggaataa gttgacaaaa gtcacgccgc aaggcttcct 30540 gaagagaaac ggcggtagcc tggatatctg caacggagcc aaaaccttca gtgtcacttc 30600 caataaacag ataaaactct aaatagtccc cacttaaaac cgaaacagcc gcagcaaagg 30660 taggacacgg tcgcacttcc tgagctctaa taaggctaaa caccacacgg cgcagttcag 30720 aaggcaaaaa gtctgtaagc tctagctggg cacacacact ctccaccaga ctcttgtgaa 30780 gtcttagaca aaaacatgct cccatagaca ctcctaaagc tgccatagta ctcacggacg 30840 gctggctgtc agaaaagagc tatgagggtg aaatgccaag cacagcgttt atatagtcct 30900 caaagtaggg cgtgtggaaa acgaaaagga atataacggg gcgtttgagg aagtggtgcc 30960 aagtacagtc ataaattgtg ggcgcgtggt aaatgttaag tgcagtttcc ctttggcggt 31020 tggcccggaa agttcacaaa aagtacagca cgt 31053 <210> 3 <211> 10129 <212> PRT <213> Artificial Sequence <220> <223> APQA1701-40P protein seq. <400> 3 Met Lys Tyr Thr Ile Val Pro Ala Pro Arg Asn Leu His Asp Tyr Val 1 5 10 15 Leu Glu Leu Leu Glu Glu Trp Gln Pro Asp Cys Leu Asp Cys Glu Tyr 20 25 30 Pro His Gly Gly Pro Ser Pro Pro Thr Leu His Asp Leu Phe Asp Val 35 40 45 Glu Leu Glu Thr Ser His Ser Pro Leu Val Gly Leu Cys Asp Ser Cys 50 55 60 Ala Glu Ala Asp Thr Asp Ser Ser Ala Ser Thr Glu Ala Asp Ser Gly 65 70 75 80 Phe Ser Pro Leu Ser Thr Pro Pro Val Ser Pro Leu Pro Pro His Pro 85 90 95 Thr Ser Pro Ala Ser Ile Ser Asp Asp Met Leu Leu Cys Leu Glu Glu 100 105 110 Met Pro Thr Phe Asp Asp Glu Asp Glu Val Arg Ser Ala Ala Thr Thr 115 120 125 Phe Glu Arg Trp Glu Asn Thr Phe Asp Pro His Val Gly Pro Ile Phe 130 135 140 Gly Cys Leu Arg Cys Ala Phe Tyr Gln Glu Gln Asp Glu Asn Ala Leu 145 150 155 160 Cys Gly Leu Cys Tyr Leu Lys Ala Leu Ala Glu Pro Ala Ser Ala Gly 165 170 175 Ala Glu Glu Glu Asp Asp Glu Val Ile Phe Val Ser Ala Lys Pro Gly 180 185 190 Gly Arg Lys Arg Ser Ala Ala Thr Pro Cys Glu Pro Asp Gly Val Ser 195 200 205 Lys Arg Pro Cys Val Pro Glu Pro Glu Gln Thr Glu Pro Leu Asp Leu 210 215 220 Ser Leu Lys Pro Arg Pro His Met Asp Pro Leu Lys Ile Cys Glu Asn 225 230 235 240 Tyr Leu Thr Phe Arg Ala Ile Ile Arg Gly Ser Thr Leu Ser Pro Gly 245 250 255 Phe Phe Arg Arg Trp Cys Phe Pro Ala Leu Ala Asp Val Val Gly Asn 260 265 270 Ile Val Glu Gln Glu Glu Gly Arg Phe Trp Gln Ile Leu Pro Glu Asn 275 280 285 His Thr Phe Trp Gly Leu Leu Arg Arg Gly Phe Thr Val Ala Ser Phe 290 295 300 Thr Glu Ile Ile Thr Ala Ala Gln Leu Glu Asn Arg Gly Arg Gln Leu 305 310 315 320 Ala Phe Leu Ala Phe Ile Ser Phe Leu Leu Arg Asn Trp Pro Ser Asp 325 330 335 Ser Val Val Pro Glu Ala Asp Arg Leu Asp Leu Val Cys Ala Pro Ala 340 345 350 Trp Ser Gln Thr Ala Arg Leu Ile Asn Asp Leu Gln Asp Ser Val Leu 355 360 365 Glu Glu Gln Gly Ser Ala Glu Glu Glu Glu Cys Glu Glu Ala Leu Leu 370 375 380 Ala Gly Asp Ser Asp Asp Pro Leu Phe Gly Met Glu Gln Asn Ala Asp 385 390 395 400 Met Glu Pro Asp Arg Gln Val Asn Gln Arg Pro Pro Arg Phe Arg Ala 405 410 415 Arg Gly Ala Gly Val Arg Gly Arg Gly Arg Val Arg Arg Ser Ala Phe 420 425 430 Ser Arg Gly Gln Arg Arg Pro Ile Ile Arg Val Asp Asp Leu Gln Leu 435 440 445 Pro Asp Pro Leu Tyr Val Met Gln Ala Leu Gln Arg Asp His Thr Leu 450 455 460 Glu Met Pro Arg Gly Gln Val Asp Phe Ser Trp Ile Glu Ala Glu Glu 465 470 475 480 Arg Arg Val Gly Pro Thr Asp Glu Trp Tyr Phe Glu Ala Val Lys Thr 485 490 495 Tyr Lys Ala Lys Pro Gly Asp Asp Leu Gln Thr Ile Ile Lys Asn Tyr 500 505 510 Ala Lys Ile Ser Leu Glu Cys Gly Ala Val Tyr Glu Ile Asn Ser Lys 515 520 525 Ile Arg Val Thr Gly Ala Cys Tyr Ile Ile Gly Asn Cys Ala Val Leu 530 535 540 Arg Pro Asn Leu Pro Ala Gly Glu Ala Met Phe Glu Val Leu Asn Val 545 550 555 560 Asp Phe Ile Pro Ser Ile Gly Phe Met Glu Arg Ile Val Phe Ser Asn 565 570 575 Val Ile Phe Asp Cys Arg Thr Thr Ala Thr Val Val Cys Cys Ile Ser 580 585 590 Glu Arg Asn Thr Leu Phe His Asn Cys Val Phe Ser Gly Pro His Met 595 600 605 Leu Cys Leu Asp Leu Arg Ala Gly Ala Glu Val Arg Gly Cys His Phe 610 615 620 Val Gly Ala Val Cys Ala Leu Arg Ser Lys Gly Leu Tyr Ser Ile Arg 625 630 635 640 Val Lys Asn Ser Ile Phe Glu Lys Cys Ala Phe Gly Val Val Thr Gly 645 650 655 Ser Lys Ala Ser Ile Ser His Cys Met Phe Lys Asp Cys Thr Cys Ser 660 665 670 Ile Met Leu Gly Gly Gln Gly Thr Ile Ala His Ser Gln Phe Ile Val 675 680 685 Thr Thr Ser Ala Glu Ala Pro Met Asn Leu Gln Leu Cys Thr Cys Glu 690 695 700 Gly Asn Gly Ser His Val Val Pro Leu Gly Asn Ile His Phe Ala Ser 705 710 715 720 His Arg Glu Ala Ser Trp Pro Thr Phe Tyr Ala Asn Thr Leu Val Arg 725 730 735 Val Arg Leu Tyr Met Gly Arg Arg Arg Gly Val Phe His Pro Lys Gln 740 745 750 Ser Thr Leu Ser Met Cys Val Ile Ala Ala Pro Arg Gly Val Val Gln 755 760 765 Arg Ile Tyr Leu Phe Gly Val Tyr Asp Ala Thr Cys Ala Ile Met Gln 770 775 780 Leu Gly Glu Ala Gly Asn Ala Ala Ser Glu Arg Leu Cys Thr Cys Gly 785 790 795 800 Phe Arg His Ser Thr Pro Ser Leu Arg Ala Thr Tyr Val Thr Asp Thr 805 810 815 Arg Ile Asp Arg Glu Leu Asn Ser Gln Asp Thr Ala Glu Phe Phe Ser 820 825 830 Ser Asp Glu Asp Asn Phe Met Asp Pro Gln Gln Lys Gly Leu Val Asn 835 840 845 Thr Cys Phe Val Thr Thr Arg Ile Pro Ser Trp Ala Gly Ala Arg Gln 850 855 860 Asn Val Thr Gly Ser Asp Leu Glu Gly Lys Pro Val Pro Ser Asp Val 865 870 875 880 Leu Glu Ser Gly Arg Pro Leu Ala Ala Pro Arg Ile Arg Thr Leu Tyr 885 890 895 Glu Glu Gln Gln Leu Asn Met Leu Ala Val Asn Val Leu Leu Asp Glu 900 905 910 Leu Lys Ile Gln Val Ala Ala Met Gln Asn Ser Val Thr Ala Ile Gln 915 920 925 Arg Glu Val Asn Asp Leu Lys Gln Arg Ile Ala Arg Asp Met Glu Glu 930 935 940 Lys Ala Gly Leu Gly Arg Leu Gln Asn Gln Gln Asp Glu Pro Ala Gln 945 950 955 960 Asp Pro His Gln Arg Ala Gly Pro Arg Ala Thr Phe His Ser Asp Arg 965 970 975 Asn His Pro Asn Lys Glu Ala Glu Ala Met Glu Gly Gln Asn Pro Ala 980 985 990 Cys Ser Arg His Ala Ser Ser His Ser Ile Gln Pro Gln Ala Ser Lys 995 1000 1005 Pro Lys Lys His Arg Asn Tyr Leu Asp Gly Ala Ala Val Asp Asp Leu 1010 1015 1020 Lys Ser Leu Trp Asp Arg Leu Gln Thr Leu Gln Gln Ser Leu Thr Asn 1025 1030 1035 1040 Met Pro Tyr Ala Glu Gly Leu Lys Pro Leu Lys Asn Phe Ala Ser Phe 1045 1050 1055 Glu Glu Leu Leu Ser Met Gly Gly Asp Ser Leu Leu Asn Asp Leu Leu 1060 1065 1070 Asp Ile Gln Asp Ser Ile Thr Gln Cys Cys Met Lys Val Ser Lys Tyr 1075 1080 1085 Leu Asn Lys Asp Gly Ser Cys Ala Ser Leu Asn Tyr Tyr Lys Gln Pro 1090 1095 1100 Phe Ile Ala Ala Val Tyr Gly Pro Thr Gly Cys Gly Lys Ser Gln Leu 1105 1110 1115 1120 Leu Arg Asn Leu Met Ser Ala Gln Leu Ile Val Pro Ala Pro Glu Thr 1125 1130 1135 Val Phe Phe Ile Thr Pro Gln Val Asp Met Ile Pro Pro Gln Glu Ile 1140 1145 1150 Ala Ala Trp Glu Thr Gln Ile Cys Glu Gly Asn Phe Leu Ala Gly Pro 1155 1160 1165 Glu Asn Thr Val Val Pro Gln Ser Gly Ser Ile Met Pro Lys Phe Val 1170 1175 1180 Gln Met Ser Tyr Ala Glu Leu Thr Asn Glu Ala Asn Tyr Asp Val Thr 1185 1190 1195 1200 Asn Pro Thr Asn Val Phe Ala Arg Ala Ala Ser Lys Gly Pro Leu Ala 1205 1210 1215 Ile Val Met Asp Glu Cys Met Glu Asp Leu Gly Gly Asn Arg Gly Ile 1220 1225 1230 Ala Lys Phe Phe His Ala Phe Pro Ser Lys Leu Leu Asp Arg Phe Pro 1235 1240 1245 Lys Cys Thr Gly Tyr Ser Val Ile Val Val Leu His Asn Met Asn Pro 1250 1255 1260 Arg Arg Asp Gln Gly Gly Asn Ile Ala Asn Leu Lys Ile Gln Ala Lys 1265 1270 1275 1280 Met His Ile Ile Ser Pro Lys Val His Pro Ser Gln Leu Asn Arg Phe 1285 1290 1295 Ile Asn Ile Tyr Thr Lys Gly Gln Pro Thr Ala Ile Ser Leu Leu Leu 1300 1305 1310 Lys Asp Ile Phe Asn Tyr His Arg Leu Asn Thr Asn Phe Asp Trp Ile 1315 1320 1325 Val Tyr Asn Thr Glu Pro Ile Asp Asn Cys Met His Trp Leu Tyr Leu 1330 1335 1340 Ser Pro Asp Glu Gly Leu Ile Pro Met Tyr Leu Asn Ile Gln Ala Lys 1345 1350 1355 1360 Val Tyr Gln Ala Met Glu Arg Ile His Arg Thr Ile Thr Asp Arg Gln 1365 1370 1375 Arg Trp Thr Arg Tyr Tyr His Ser Lys Lys Lys Met Ser Leu Val Gln 1380 1385 1390 Gly His Gly Thr Ser Gly Leu Phe Thr Glu Pro Pro Asn Pro Ile Asn 1395 1400 1405 Gln Gln Glu Ser Ser Gly Pro Ser Leu Pro Ala Gln Asp Ala Ala Gln 1410 1415 1420 Ala Phe Ala Ser Ser Pro Arg Ala Gly Ala Thr Ser Thr Ile Val Asn 1425 1430 1435 1440 Pro Pro Lys Arg Lys Tyr Lys Gly Ala Val Val Val Gln Arg Ala Thr 1445 1450 1455 Leu Ser Ile Ser Ala Val Leu Asp Asn Gly Gln Cys Val Glu Ile Lys 1460 1465 1470 Tyr His Ser Asn Leu Ala Ser Ala Leu Thr Asn Leu Cys Asn Thr Asn 1475 1480 1485 Leu Tyr Asp Leu Pro Ala Cys Leu Asn Arg Pro Ile Thr Ala His Asn 1490 1495 1500 Leu Pro Ala Leu Ile Glu Glu Ala Ala Ala Pro Tyr Ser Leu Ile Cys 1505 1510 1515 1520 Tyr Tyr Gln Arg Gly Thr Val Arg Arg Val Glu Phe Gln Ala Glu Val 1525 1530 1535 Pro Leu Leu Ser Phe Pro Leu Lys Phe Leu Val Lys Gln Gly Lys Val 1540 1545 1550 Phe Leu Ile Lys Asp Ile Ser Gln Met Gln Lys Cys Glu Phe Cys Gly 1555 1560 1565 Ser Phe Phe Lys Val Thr His Thr Cys Ala Leu Arg Arg Arg Asp Phe 1570 1575 1580 Tyr Phe His His Val Ala Ala His Ser Ala Asp Trp Trp Glu Lys Ile 1585 1590 1595 1600 Ser Phe Thr Pro Ile Gly Ala Pro Pro Asn Thr Glu Arg Leu Phe Ile 1605 1610 1615 Val Tyr Asp Val Glu Thr Tyr Thr Trp His Gly Lys Phe Gly Lys Gln 1620 1625 1630 Leu Val Pro Phe Met Leu Val Phe Gln Leu Leu Gly Asp Asp His Leu 1635 1640 1645 Val Asn Val Ala Lys Thr Leu Ala Thr Glu Gln Asn Trp Glu Ile Trp 1650 1655 1660 Asn Gly Lys Glu Gln Asp Thr Leu Tyr Tyr Cys Ile Thr Pro Glu Lys 1665 1670 1675 1680 Arg Ala Ile Gly Val Lys Phe Lys Lys Phe Arg Asp Thr Leu Gln Gln 1685 1690 1695 His Ile Ala Ala Ser Leu Trp Ser His Val Ile Cys Gln Asn Pro Gln 1700 1705 1710 Leu Gln Glu Lys Ala Thr Thr Leu Gly Leu Glu Ser Pro Glu Glu Leu 1715 1720 1725 Thr Pro Asp Gln Leu Lys Lys Phe Lys Leu Lys Gly Asn Pro Arg Phe 1730 1735 1740 Ile Glu Val Tyr Ala Val Gly His Asn Ile Thr Gly Phe Asp Glu Ile 1745 1750 1755 1760 Leu Leu Ala Ala Gln Val Val Ser Thr Arg Ala Glu Ile Pro Pro Val 1765 1770 1775 Phe Glu Ile Cys Arg Asn Phe Met Pro Arg Ala Gly Arg Leu Leu Phe 1780 1785 1790 Asn Asp Ile Thr Tyr Ser Leu Pro Asn Pro Ser Tyr Val Pro Ala Lys 1795 1800 1805 Ser Tyr Glu His Trp Glu Gln Gly Gln Val Leu Ala Ser Asp Leu Lys 1810 1815 1820 Ser Gln Tyr Ile Lys Phe Met Val Arg Asp Thr Phe Ser Leu Thr His 1825 1830 1835 1840 Thr Ser Leu Lys Asn Ala Ala Lys Ala Tyr Ser Leu Thr Val Ser Lys 1845 1850 1855 Gly Cys Cys Pro Tyr Gln Ala Val Asn Glu Phe Tyr Met Leu Gly Ser 1860 1865 1870 Tyr Gln Gln Asp Ala Asp Gly Phe Pro Asp Leu Lys Tyr Trp Lys Asp 1875 1880 1885 Gln Glu Glu Tyr Ser Phe Asn Lys Asp Leu Trp Ile Lys Glu Lys Lys 1890 1895 1900 Gly Ala Tyr Asp Ile Ile Gln Gln Thr Leu Asp Tyr Cys Ala Leu Asp 1905 1910 1915 1920 Val Gln Val Thr Ala Gln Leu Val Asn Lys Leu Ile Glu Ser Tyr Gln 1925 1930 1935 Ile Phe Ile Lys Asn Ser Val Asn Leu Pro Glu Thr Ser Phe Asn Val 1940 1945 1950 Phe Gln Arg Pro Thr Ile Ser Ser Asn Ser His Ala Ile Phe Lys Gln 1955 1960 1965 Ile Leu Tyr Lys Ala Glu Lys Pro Asn Thr His His Leu Ser Thr Ile 1970 1975 1980 Leu Met Ala Pro Ser Asn Glu Met Tyr Glu Tyr Val Arg Leu Ser Ile 1985 1990 1995 2000 Arg Gly Gly Arg Cys Tyr Pro Thr Tyr Ile Gly Val Leu Gln Glu Pro 2005 2010 2015 Val Phe Val Tyr Asp Ile Cys Gly Met Tyr Ala Ser Ala Leu Thr His 2020 2025 2030 Pro Phe Pro Ala Gly Ser Pro Leu Asn Pro Tyr Glu Arg Ala Leu Ala 2035 2040 2045 Ile Lys Ala Tyr Glu Gln Lys Met Leu Asn His Lys Thr Ile Ser Tyr 2050 2055 2060 Phe Asp Lys Asp Leu Leu Pro Gly Ile Phe Thr Ile Asp Ala Asp Pro 2065 2070 2075 2080 Pro Ala Glu Glu Phe Leu Asp Val Leu Pro Phe Cys Ser Arg Lys 2085 2090 2095 Gly Gly Arg Leu Cys Trp Thr Asn Glu Pro Leu Arg Gly Glu Ile Ala 2100 2105 2110 Thr Ser Ile Asp Val Ile Thr Leu His Asn Arg Gly Trp Lys Val Thr 2115 2120 2125 Leu Ile Pro Asp Thr Arg Thr Thr Val Phe Pro Glu Trp Lys Cys Leu 2130 2135 2140 Ala Arg Glu Tyr Val Gln Leu Asn Ile Ser Ala Lys Glu Glu Ala Asp 2145 2150 2155 2160 Lys Ser Lys Asn Gln Thr Met Arg Ser Ile Ala Lys Leu Leu Ser Asn 2165 2170 2175 Ala Leu Tyr Gly Ser Phe Ala Thr Lys Leu Asp Asn Lys Lys Thr Val 2180 2185 2190 Phe Ser Asp Gln Ile Glu Ser Asn Ile Ala Lys Glu Ile Ala Ser Gly 2195 2200 2205 Ala Tyr Val Val Lys Ser Ser Ser Tyr Ile Glu Thr Asp Asn Leu Cys 2210 2215 2220 Ala Glu Ile Met Pro Glu Phe Val Val Ala Tyr Pro Pro Val Asn Ser 2225 2230 2235 2240 Asp Val Arg Gln Leu Ala Pro Pro Ser Cys Ser Glu Glu Asp Pro Thr 2245 2250 2255 Lys Asp Pro Leu Ala Glu Ala Pro Phe Met His Asn Phe Ser Met Thr 2260 2265 2270 Ser Tyr His Tyr Lys Pro Ile Met Phe Ile Asp Ala Glu Asp Asp Asp 2275 2280 2285 Phe Cys Leu His Thr Leu Glu Lys Ser Thr Pro Leu Ile Ala Asn Asn 2290 2295 2300 Arg Tyr Pro Ser Gln Ile Ala Ser Phe Val Leu Ala Trp Thr Arg Ala 2305 2310 2315 2320 Phe Val Ser Glu Trp Ser Gln Phe Leu Tyr Glu Asn Asp Ala Gly Thr 2325 2330 2335 Pro Leu Glu Asn Arg Val Leu Lys Ser Val Tyr Gly Asp Thr Asp Ser 2340 2345 2350 Leu Phe Thr Thr Met Glu Gly Tyr Lys Leu Met Glu Glu Lys Gly Lys 2355 2360 2365 Lys Arg Leu Lys Lys Asn Gly Gly Lys Leu Val Phe Asp Pro Ser Asn 2370 2375 2380 Pro Glu Leu Thr Trp Leu Val Glu Cys Glu Thr Gln Cys Glu Lys Cys 2385 2390 2395 2400 Gly Ser Asp Ala Tyr Ser Ser Glu Ser Val Tyr Leu Ala Pro Lys Leu 2405 2410 2415 Tyr Ala Leu Lys Asp Thr Thr Cys Pro Lys Cys His His Val Gly Lys 2420 2425 2430 Gly Lys Leu Arg Ala Lys Gly His Ala Thr Ser Thr Leu Ser Tyr Asp 2435 2440 2445 Val Leu Lys Ala Cys Tyr Tyr Ala Asp Met Gln Gln Gly Ser Asp Val 2450 2455 2460 Phe Lys Thr Ser Arg Met Ser Leu Arg Arg Thr Leu Thr Ser Val Gln 2465 2470 2475 2480 Ala His Val Gln Pro Phe Thr Val Thr Glu Thr Thr Leu Thr Arg Lys 2485 2490 2495 Leu Arg Pro Trp Lys Asp Lys Thr Leu His Ala Leu Asp Met His Arg 2500 2505 2510 Leu Ile Pro Tyr Ser Arg Lys His Pro Asn Pro Arg Asn Thr Glu Thr 2515 2520 2525 Thr Trp Met Glu Leu Gln Trp Met Thr Met Ser Leu Asn Ala Leu Asp 2530 2535 2540 Cys Ala Arg Leu Thr Gly Gln Thr Pro Tyr Thr Val Glu Val Phe Arg 2545 2550 2555 2560 Pro Leu Arg Asn Ile Phe Asn Arg Val Arg Glu Tyr Thr Arg Ala Ser 2565 2570 2575 Thr Thr Ser Val Gly Leu Ala Trp Met Ser Lys Tyr Ile Tyr Gln Tyr 2580 2585 2590 His Arg Leu Met Leu Met Asn Leu Ser Pro Arg Glu Pro Ala Thr Glu 2595 2600 2605 Gly Trp Pro Leu Phe Trp Tyr Pro Pro His Leu Leu Val Gly Tyr 2610 2615 2620 Gln Tyr Leu Val Arg Thr Cys Asn Asp Tyr Val Phe Asp Thr Arg Ser 2625 2630 2635 2640 Tyr Ser Arg Leu Lys Tyr Thr Glu Ile His Leu Pro Leu Gln Gln Lys 2645 2650 2655 Leu Asn Trp Thr Val Met Ala Asn Cys Ser Tyr Thr Ile Asn Thr Gly 2660 2665 2670 Ala Tyr His Arg Phe Ile Asp Phe Glu Asn Phe Ala Glu Thr Leu Ala 2675 2680 2685 Gln Val Gln Gln Ala Val Leu Ala Glu Arg Val Val Ala Asp Leu Ala 2690 2695 2700 Leu Ile Arg Pro Leu Arg Gly Tyr Gly Thr Thr Asn Met Ala Gly Asp 2705 2710 2715 2720 Arg Gln Val Pro Val Glu Gly Leu Leu Gln Asp His Tyr Lys Asn Leu 2725 2730 2735 Ser Gln Cys Gln Asn Glu Ala Trp Gly Leu Ala Asp Arg Met Arg Ile 2740 2745 2750 Gln Gln Ala Gly Asn Lys Asp Leu Val Ile Leu Thr Thr Ile Arg Lys 2755 2760 2765 Leu Lys Thr Ala Phe Phe Asn Phe Leu Val Ser Pro Arg Asn Pro His 2770 2775 2780 Thr Met Leu Ser Leu Pro Cys Asp Cys Leu Trp Leu Asp Ala Phe Val 2785 2790 2795 2800 Glu Lys Phe Thr Asp Pro Glu Leu Ser Gln Phe Gln Thr Ile Gln Ser 2805 2810 2815 Leu Pro Ser Gln Ser Val Thr Lys Ser Ile Ile Ser Ala Leu Ser Leu 2820 2825 2830 Pro Arg Pro Ala Pro Cys Thr Pro Leu Ser Gly Gly Ala Phe Glu Leu 2835 2840 2845 Arg Pro Arg Glu Asn Gly Arg Ala Val Thr Glu Glu Met Arg Arg Arg 2850 2855 2860 Arg Gly Glu Val Ile Glu Arg Phe Ile Asp Arg Leu Pro Val Arg Arg 2865 2870 2875 2880 Arg Arg Arg Arg Ala Pro Pro Pro Pro Val Ser Glu Glu Leu Ser 2885 2890 2895 Glu Pro Glu Ala Glu Ala Ser Pro Pro Pro Ala Ser Pro Pro Arg 2900 2905 2910 Arg Ser Phe Glu Glu Glu Val Arg Asp Thr Ile Val Glu Ala Ile Arg 2915 2920 2925 Leu Leu Gln Glu Glu Leu Thr Val Ser Ala Arg Asn Glu Gln Phe Phe 2930 2935 2940 Asn Phe Ala Val Asp Phe Tyr Glu Val Ile Gln Arg Leu Glu Met Leu 2945 2950 2955 2960 Gly Asn Ile Asn Glu Leu Thr Ile Arg Arg Trp Val Met Tyr Phe Phe 2965 2970 2975 Val Ala Glu His Val Ala Thr Thr Leu Asn Tyr Leu His His Asn Leu 2980 2985 2990 Arg Leu Tyr Pro Pro Cys Ser Arg Trp Val Asp Leu Glu Leu Ala Gln 2995 3000 3005 Val Val Met Arg Ala Arg Asp His Glu Gly Gln Val Val Tyr Ser Arg 3010 3015 3020 Val Trp Asn Glu Met Gly Glu Asn Ala Phe Ser Gln Val Met Ala Arg 3025 3030 3035 3040 Val Ser Gly Asp Leu Ala Ala Thr Val Glu Arg Ala Gly Leu Gly Glu 3045 3050 3055 Leu Glu Glu Glu Glu Met Glu Gln Phe Met Ala Asp Ile Ala Tyr His 3060 3065 3070 Glu Asn Ser Gly Asp Val Ser Glu Ile Leu Arg Gln Val Ala Ile Asn 3075 3080 3085 Asp Thr Glu Val Asp Ser Met Glu Leu Ser Phe Arg Phe Lys Val Thr 3090 3095 3100 Gly Pro Val Val Phe Ser Gln Asn Arg Gln Ile Gln Ser Ile Asn Arg 3105 3110 3115 3120 Arg Val Val Ala Leu Ala Ser Gln Leu Arg Met Gln His Arg Pro Leu 3125 3130 3135 Pro Ala His His Glu Gln Val Gln Leu Pro Met His Pro Val Leu 3140 3145 3150 Arg Gln Met Lys Pro Leu Pro Ala Ser Ala Ser Lys Leu Glu Ala Glu 3155 3160 3165 Ser Glu Gly Leu Ala Arg Leu Gln Gly Gly Ala Ala Pro Glu Leu His 3170 3175 3180 Pro Arg Val Gln Met Lys Gln Glu Ala Ala Glu Ala Tyr Ile Pro Ala 3185 3190 3195 3200 Ser Asn Val Phe Arg Asp Asn Glu Gly Glu Glu Ala Glu Gly Leu Arg 3205 3210 3215 His Leu Lys Tyr Glu Ser Gly Arg Leu Leu Arg Asn Asp His Thr Ser 3220 3225 3230 Lys Arg Val Leu Gly Glu Arg Asp Phe Glu Lys Asp Pro Gln Asn Gly 3235 3240 3245 Ile Ser Ala Ala Glu Ala His Leu Lys Ser Ala Asp Leu Val Thr Ala 3250 3255 3260 Tyr Glu His Thr Val Lys Ala Glu Val Asn Phe Gln Thr Thr Phe Asn 3265 3270 3275 3280 Asn Asn Val Arg Thr Leu Ile Ala Arg Glu Glu Val Val Ile Gly Leu 3285 3290 3295 Met His Leu Trp Asp Phe Val Glu Ala Phe Leu Glu Asn Pro Val Ser 3300 3305 3310 Lys Ala Leu Thr Ala Gln Leu Phe Leu Ile Val Gln His Cys Arg Asp 3315 3320 3325 Glu Gly Val Leu Arg Glu Ser Leu Leu Asn Ile Ala Glu Pro Glu Ser 3330 3335 3340 Arg Trp Leu Val Asp Leu Leu Asn Leu Leu Gln Thr Ile Val Val Gln 3345 3350 3355 3360 Glu Arg Gly Leu Ala Val Gly Glu Lys Val Ala Ala Ile Asn Tyr Ser 3365 3370 3375 Val Ile Thr Leu Ser Lys His Tyr Ala Arg Lys Ile Phe Asn Ser Val 3380 3385 3390 Phe Val Pro Ile Asp Lys Glu Ala Lys Ile Asn Thr Phe Tyr Met Arg 3395 3400 3405 Thr Val Val Lys Leu Leu Val Leu Ser Asp Asp Leu Gly Met Tyr Arg 3410 3415 3420 Asn Glu Arg Ile Glu Arg Ala Val Ser Gly Ala Arg Gln Arg Glu Leu 3425 3430 3435 3440 Asn Asp Arg Glu Leu Met His Arg Leu Arg Gln Ala Leu Ala Ser Asn 3445 3450 3455 Gly Leu Pro Glu Leu Glu Gly Glu Asp Ser Ala Asn Ile Arg Ser Lys 3460 3465 3470 Glu Glu Trp Gly Val Gly Ala Gly Gly Gly Val Ala Ser Ala Arg Tyr 3475 3480 3485 Pro His Leu Leu Asp Tyr Glu Glu Glu Glu Asn Pro Asp Gly Ser Val 3490 3495 3500 Ser Phe Gln Gln His Glu Arg Gly Ala Gln Pro His Glu Asn Gly Gly 3505 3510 3515 3520 His Ala Glu Ser Ala Tyr Ser Arg Arg Gln Leu Gly Arg Phe Tyr Met 3525 3530 3535 Ser Ala Ala Leu Asn Pro Met Lys Met Ala Ala Met Gln Ser Gln Pro 3540 3545 3550 Thr Ala Asp Asp Ser Trp Ala Ala Ser Ile Ser Arg Ile Met Ser Leu 3555 3560 3565 Thr Ala Gly Asp Arg His Lys Phe Ser Ser Gln Pro Phe Ala Asn Arg 3570 3575 3580 Leu Asp Ala Ile Leu Glu Ala Val Val Pro Ser Arg Lys Asp Pro Thr 3585 3590 3595 3600 His Glu Lys Val Leu Thr Ile Val Asn Ala Leu Ile Glu Asn Gly Ala 3605 3610 3615 Ile Arg Arg Asp Glu Gly Ala Gly Val Tyr Asp Ala Leu Leu His Arg 3620 3625 3630 Cys Ala Lys Tyr Asn Ser Leu Asn Ala Gln Ser Asn Leu Glu Arg Leu 3635 3640 3645 Ala Gly Asp Val Arg Glu Ala Val Ala Gln Gln Val Arg Ile Ala Thr 3650 3655 3660 Gly Asn Leu Gly Ser Leu Thr Ala Leu Asn Gly Phe Leu Ala Arg Leu 3665 3670 3675 3680 Pro Ala Asn Val Glu Arg Gly Gin Glu Asn Tyr Thr Gly Phe Val Ser 3685 3690 3695 Ala Leu Lys Leu Leu Val Ser Glu Val Pro Ser Thr Glu Val Tyr Gln 3700 3705 3710 Ser Gly Pro His Tyr Phe Leu Gln Ser Ser Arg Asn Gly Thr Gln Thr 3715 3720 3725 Val Asn Leu Thr Asn Ala Phe Glu Asn Leu Lys Pro Leu Trp Gly Val 3730 3735 3740 Lys Ala Pro Thr Met Glu Arg Leu Ser Ile Ser Ala Leu Leu Thr Pro 3745 3750 3755 3760 Asn Thr Arg Leu Leu Leu Leu Leu Val Ser Pro Phe Thr Asp Ser Val 3765 3770 3775 Ser Ile Ser Arg Asp Ser Tyr Leu Gly Tyr Leu Leu Thr Leu Tyr Arg 3780 3785 3790 Glu Ala Leu Gly Arg Asn His Leu Asp Glu Arg Thr Leu Glu Glu Val 3795 3800 3805 Thr Glu Val Ser Arg Ala Met Gly Ser Glu Asn Ile Asn Asn Leu Gln 3810 3815 3820 Ala Thr Leu Asn Phe Leu Leu Thr Asn Arg Gln Lys Arg Ile Pro Lys 3825 3830 3835 3840 Asp Tyr Ser Leu Thr Pro Glu Glu Glu Arg Ile Val Arg Phe Val Gln 3845 3850 3855 Gln Ala Val Ser Leu Arg Met Met Gln Glu Asn Leu Ser Pro Thr Glu 3860 3865 3870 Ala Leu Asp Val Thr Ala Ala Asn Met Glu Pro Ser Phe Tyr Ala Ser 3875 3880 3885 Asn Arg Asp Phe Ile Asn Lys Leu Met Asp Tyr Phe His Arg Ala Ala 3890 3895 3900 Ala Met Ala Pro Asp Tyr Phe Leu Gly Ala Val Met Asn Pro Arg Trp 3905 3910 3915 3920 Leu Pro Pro Glu Gly Phe Phe Thr Gly Val Phe Asp Phe Pro Glu Arg 3925 3930 3935 Asp Ser Tyr Thr Trp Asp Gly Leu Asp Ser Ser Leu Glu Leu Thr Arg 3940 3945 3950 Gln Asp Ala Met Arg Phe Leu Glu Asp Lys Phe Met Asp Asp Asp Gln 3955 3960 3965 Arg Thr Glu Ser Arg Ser Leu Ser Arg Val Thr Thr Pro Ala Ser Ser 3970 3975 3980 Arg Arg Ser Ser Val Thr Met Pro Pro Gly Gly Phe Met Gly Met Ile 3985 3990 3995 4000 Asn Asn Asn Lys Asn Asp Asn Ser Leu Arg Glu Ile Asp Gly Leu Ala 4005 4010 4015 Asp Lys Leu Ala Arg Trp Lys Thr Tyr Lys Arg Glu Ser Glu Glu Ala 4020 4025 4030 Arg Val Ser Leu Pro Ala Val Val Arg Pro Lys Gln Tyr Arg Pro Arg 4035 4040 4045 Ser Pro Ile Ser Ser Asp Asp Ser Asp Asp Gly Met Ser Arg Pro Asp 4050 4055 4060 Pro Phe Leu Lys Phe Glu Gly Ser Gly Asn Pro Phe Ala His Leu Arg 4065 4070 4075 4080 Pro Lys Leu Gly Arg Gly Leu Met Glu Phe Ser Ser Ser Pro Pro Pro 4085 4090 4095 Ser Tyr Glu Thr Val Met Ala Gln Val Pro Ser Ile Leu Ala Pro Leu 4100 4105 4110 Val Pro Pro Arg Tyr Lys Gly Ala Thr Glu Gly Arg Asn Ser Ile Arg 4115 4120 4125 Tyr Ser Gln Leu Pro Pro Leu Phe Asp Thr Thr Lys Leu Tyr Leu Ile 4130 4135 4140 Asp Asn Lys Ser Ser Asp Ile Gln Ala Leu Asn Tyr Gln Asn Asp His 4145 4150 4155 4160 Ser Asn Phe Leu Thr Thr Val Val Gln Asn Ala Asn Tyr Thr Pro Met 4165 4170 4175 Glu Ala Ser Thr Gln Ser Ile Gln Leu Asp Glu Arg Ser Arg Trp Gly 4180 4185 4190 Gly Glu Phe Lys Ser Ile Leu His Met Asn Met Pro Asn Val Thr Glu 4195 4200 4205 Tyr Met Phe Ser Asn Ser Phe Lys Ala Tyr Leu Pro Ala Thr Ala Asp 4210 4215 4220 Asn Phe Gly Lys Val Leu Thr Tyr Glu Trp Tyr Thr Leu Thr Ile Pro 4225 4230 4235 4240 Glu Gly Asn Tyr Ser Glu Val Met Leu Leu Asp Leu Leu Asn Asn Ala 4245 4250 4255 Val Val Glu Asn Tyr Leu Ala His Gly Arg Gln His Asn Val Arg Glu 4260 4265 4270 Glu Asp Met Gly Leu Lys Phe Asp Thr Arg Asn Phe His Leu Gly Phe 4275 4280 4285 Asp Pro Glu Thr Lys Leu Val Met Pro Gly Phe Tyr Thr Asn Glu Ala 4290 4295 4300 Phe His Pro Asp Ile Val Leu Ser Pro Gly Cys Ala Val Asp Phe Thr 4305 4310 4315 4320 His Ser Arg Leu Asn Asn Phe Leu Gly Ile Arg Lys Arg Leu Pro Tyr 4325 4330 4335 Gln Glu Gly Phe Val Ile Thr Trp Glu Asp Leu Gln Gly Gly Asn Ile 4340 4345 4350 Pro Ala Leu Leu Asp Leu Glu Asn Tyr Asn Pro Asp Ile Pro Gly Ala 4355 4360 4365 Asp Ile Thr Pro Leu Met Tyr Asp Ser Lys Gly Arg Pro Tyr His Val 4370 4375 4380 Gly Glu Asp Pro Ser Ala Gly Ser Thr Phe Thr Trp Tyr Arg Ser Trp 4385 4390 4395 4400 Phe Val Ala Tyr Asn Tyr Gly Pro Ala Asp Gly Ile Lys Ser Lys Thr 4405 4410 4415 Val Leu Val Ala Pro Asp Ile Thr Cys Gly Val Glu Gln Ile Tyr Trp 4420 4425 4430 Ser Leu Pro Asp Met Ala Val Asp Pro Val Thr Phe Thr Ser Ser His 4435 4440 4445 Asn Pro Ser Ser Tyr Pro Val Val Gly Thr Glu Leu Leu Pro Leu Leu 4450 4455 4460 Pro Arg Ser Phe Tyr Asn Gly Ser Ser Val Tyr Ser Gln Leu Leu Gln 4465 4470 4475 4480 Glu Ser Thr Ser Gln Thr Gln Val Phe Asn Arg Phe Pro Glu Asn Ala 4485 4490 4495 Ile Leu Lys Arg Pro Pro Ala Pro Thr Ile Ile Ser Ile Ser Glu Asn 4500 4505 4510 Val Pro Ala Leu Ser Asn His Gly Thr Leu Pro Leu Lys Asn Asn Ile 4515 4520 4525 Pro Gly Val Gln Arg Val Thr Ile Thr Asp Ala Arg Arg Arg Val Cys 4530 4535 4540 Pro Tyr Val Tyr Lys Ser Leu Gly Val Val Val Pro Arg Ala Leu Ser 4545 4550 4555 4560 Ser Lys Thr Phe Met Ala Ile Leu Ile Ser Pro Thr Asn Asn Thr Gly 4565 4570 4575 Trp Gly Leu Gly Thr His Lys Leu Phe Gly Gly Ala Lys Gln Lys Ser 4580 4585 4590 Asp Gln His Pro Val Tyr Val Gln Ala His Tyr Arg Ala Ser Trp Gly 4595 4600 4605 Ser Lys Gly Arg Arg Arg Arg Gln Gly Arg Ala Arg Gly Ala Pro Leu 4610 4615 4620 Asp Pro Lys Thr Glu Ala Glu Met Val Ala Thr Ile Asp Glu Val Ala 4625 4630 4635 4640 Arg Asn Gly Pro Pro Ala Ala Arg Leu Val Leu Glu Ala Ala Arg Arg 4645 4650 4655 Val Gly Ala Tyr Asn Leu Arg Arg Ala Arg Lys Leu Thr Pro Ala Gly 4660 4665 4670 Arg Ala Met Met Ala Met Arg Ala Arg Gln Met Val Lys Gln Ala Lys 4675 4680 4685 Lys Arg Lys Arg Arg Val Arg Phe Arg Gln Met Ala Gly Arg Asn Val 4690 4695 4700 Thr Leu Arg Leu Arg Val Pro Val Arg Thr Lys Ile Thr Gly Ala Gly 4705 4710 4715 4720 Arg Arg Arg Gly Arg Arg Pro Arg Gly Ile Arg Cys Gly Arg Met Arg 4725 4730 4735 Gly Gly Phe Leu Pro Ala Leu Ile Pro Leu Ile Ala Ala Ala Ile Gly 4740 4745 4750 Ala Val Pro Gly Ile Ala Ser Val Ala Leu Gln Ala Ala Arg His Met 4755 4760 4765 Ala Ala Ile Ser Arg Ala Ile Lys Gln Glu Leu Leu Glu Asp Leu Lys 4770 4775 4780 Pro Glu Ile Tyr Leu Pro Pro Lys Ser Thr Lys Arg Arg Thr Lys Val 4785 4790 4795 4800 Lys Thr Glu Glu Lys Val Asp Val Lys Thr Leu Val Lys Ala Lys Ser 4805 4810 4815 Arg Lys Arg Arg Ala Ala Lys Asp Glu Leu Glu Glu Asp Val Glu Phe 4820 4825 4830 Val Arg Arg Phe Ala Pro Arg Arg Pro Tyr Gln Trp Arg Gly Arg Arg 4835 4840 4845 Val Arg Ala Leu Pro Arg Pro Gly Val Pro Val Val Phe Thr Pro Gly 4850 4855 4860 Gln Arg Ser Gly Thr Ala Ser Lys Arg Ser Tyr Asp Glu Val Tyr Ala 4865 4870 4875 4880 Asp Glu Asp Val Leu Asp Gln Ala Gly Asn Met Ile Asn Glu Phe Ala 4885 4890 4895 Tyr Gly Lys Arg Val Arg Val Leu Thr His Lys Asn Pro Thr Pro Ser 4900 4905 4910 Gln Val Pro Ile Thr Pro Gln Glu Pro Val Ala Arg Pro Gly Glu Ala 4915 4920 4925 His Leu Leu Pro Thr Val Gln Val Leu Ala Pro Arg Gly Ser Arg Arg 4930 4935 4940 Glu Thr Met Leu Pro Val Thr Lys Ser Glu Gly Gly Asp Val Lys Val 4945 4950 4955 4960 Glu Asn Lys Gly Phe Glu Gln Ile Thr Pro Gly Leu Gly Val Gln Thr 4965 4970 4975 Val Asp Ile Lys Val Pro Val Lys Arg Lys Gly Asp Ala Glu Asp Glu 4980 4985 4990 Ile Ile Lys Arg Val Lys Met Glu Leu Glu Pro Tyr Glu Thr Thr Met 4995 5000 5005 Lys Met Glu Tyr Ser Glu Glu Pro Gln Val Glu Ala Phe Asp Thr Gly 5010 5015 5020 Ile Glu Pro Ser Ser Phe Phe Glu Val Arg Ser Gln Ala Arg Pro Ile 5025 5030 5035 5040 Ala Val Ala Arg Lys Arg Arg Thr Ala Ala Ala Ser Ala Pro Ala Val 5045 5050 5055 Glu Val Met Glu Val Gln Gln Ser Asn Pro Ala Thr Pro Ala Thr Ala 5060 5065 5070 Ala Ala Arg Thr Ala Thr Ala Leu Gly Pro Arg Leu Ala Arg Arg Pro 5075 5080 5085 Ser Arg Trp Gly Pro Ala Asn Ala Ile Phe Pro Asp Tyr Lys Tyr His 5090 5095 5100 Pro Ser Ile Thr Ala Arg Lys Ile Arg Gly Pro Lys Pro Thr Gly Lys 5105 5110 5115 5120 Ile Ser Arg Trp Gly Pro Ala Asn Ser Ile Leu Pro Glu Val Arg Leu 5125 5130 5135 His Pro Ser Met Val Ser Ala Val Thr Arg Ala Ala Pro Arg Arg Val 5140 5145 5150 Thr Lys Thr Arg Arg Arg Arg Arg Ala Arg Thr Arg Gln Ala Phe Val 5155 5160 5165 Leu Pro Ala Arg Thr Lys Thr Gly Ala Leu Leu Ser Gln Asn Val Arg 5170 5175 5180 Tyr His Pro Ser Ile Ser Leu Leu Arg Arg Ala Met Asp Ala Val Asn 5185 5190 5195 5200 Phe Ser Ile Leu Ala Pro Arg Tyr Gly Ala His Pro Met Met Ser Gly 5205 5210 5215 Trp Ser Gly Ile Gly Thr Ser Gly Met Asn Gly Gly Ala Phe Asn Trp 5220 5225 5230 Gly Gly Ile Trp Ser Gly Ile Lys Asn Phe Gly Ser Asn Val Lys Ser 5235 5240 5245 Trp Gly Ser Lys Ala Trp Asn Ser Gln Thr Gly Lys Leu Leu Arg Gln 5250 5255 5260 Lys Leu Asn Asp Thr Lys Val Arg Glu Lys Leu Val Glu Gly Ile Ser 5265 5270 5275 5280 Thr Gly Val His Gly Ala Leu Asp Ile Ala Asn Gln Glu Leu Ala Lys 5285 5290 5295 Gln Ile Glu Arg Arg Leu Glu Arg Gln Gln Pro Leu Glu Pro Glu Val 5300 5305 5310 Glu Val Glu Glu Glu Val Val Asp Ile Lys Pro Glu Ala Gln Ala Pro 5315 5320 5325 Leu Val Val Gln Ile Pro Lys Lys Arg Pro Arg Asp Glu Asp Leu Leu 5330 5335 5340 Ile Thr Ala Asp Glu Pro Ser Tyr Glu Glu Ser Ile Lys Thr Met 5345 5350 5355 5360 Ala Pro Leu Met Pro Met Thr Arg Pro His Pro Ser Met Ala Lys Pro 5365 5370 5375 Val Leu Val Asp Arg Pro Thr Thr Leu Glu Leu Lys Pro Ser Asp Gln 5380 5385 5390 Pro Pro Ala Tyr Ser Pro Pro Ala Pro Ser Ala Val Arg Val Thr Val 5395 5400 5405 Pro Ser Asn Ile Pro Val Val Thr Pro Ala Arg Ser Arg Gly Trp Gln 5410 5415 5420 Gly Thr Leu Ala Asn Ile Val Gly Val Gly Leu Ser Asn Val Lys Arg 5425 5430 5435 5440 Arg Arg Cys Phe Met Ala Thr Pro Ser Met Leu Pro Gln Trp Ser Tyr 5445 5450 5455 Met His Ile Ala Gly Gln Asp Ala Ala Glu Tyr Leu Ser Pro Ala Leu 5460 5465 5470 Val Gln Phe Ala Gln Ala Thr Ser Ser Tyr Phe Lys Leu Asp Asn Lys 5475 5480 5485 Phe Arg Asn Pro Thr Val Ala Pro Thr His Asp Val Thr Thr Glu Arg 5490 5495 5500 Ser Gln Arg Leu Gln Leu Arg Phe Val Pro Val Met Gln Glu Asp Gly 5505 5510 5515 5520 Gln Tyr Thr Tyr Lys Thr Arg Phe Gln Leu Ala Val Gly Asp Asn Arg 5525 5530 5535 Val Leu Asp Met Ala Ser Thr Tyr Phe Asp Ile Arg Gly Thr Leu Asp 5540 5545 5550 Arg Gly Pro Ser Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ala Leu 5555 5560 5565 Ala Pro Lys Ala Gly Ala Asn Asn Cys Leu Phe Asn Gly Gln Gly Ala 5570 5575 5580 Asn Ile Asn Thr Leu Ala Gln Val Pro Ser Ala Gly Ala Ile Thr Val 5585 5590 5595 5600 Asn Gly Gln Ala Ala Val Thr Asn Asn Thr Tyr Gln Pro Glu Pro Gln 5605 5610 5615 Leu Gly Pro Glu Ser Trp Val Asp Gly Ser Leu Ala Glu Leu Gly Asp 5620 5625 5630 Ala Ser Gly Arg Ala Leu Lys Ala Ser Thr Pro Arg Met Pro Cys Tyr 5635 5640 5645 Gly Ser Tyr Ala Pro Thr Asn Glu Asn Gly Gly Gln Ala Thr Gly 5650 5655 5660 Pro Val Glu Ser Arg Phe Tyr Lys Val Thr Thr Asn Asn Asn Asn Glu 5665 5670 5675 5680 Ala Asp Ala Met Leu Tyr Thr Glu Asp Val Asn Leu Gln Ala Pro Asp 5685 5690 5695 Thr His Leu Val His Gln Val Pro Glu Gly Gln Val Thr Gly Val Gln 5700 5705 5710 Gly Leu Gly Gln Gln Ala Ala Pro Asn Arg Pro Asn Tyr Ile Gly Phe 5715 5720 5725 Arg Asp Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Asn Gly Asn Leu 5730 5735 5740 Gly Val Leu Ala Gly Gln Ser Ser Gln Leu Asn Ala Val Val Asp Leu 5745 5750 5755 5760 Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ala Leu 5765 5770 5775 Thr Asp Arg Ser Arg Tyr Phe Ser Met Trp Asn Gln Ala Val Asp Ser 5780 5785 5790 Tyr Asp Gln Asp Val Arg Ile Ile Asp Asn His Gly Val Glu Asp Asp 5795 5800 5805 Met Pro Asn Tyr Cys Tyr Pro Leu Ser Gly Met Gly Pro Leu Thr Asn 5810 5815 5820 Met Thr Thr Met Lys Val Asn Asn Gln Asn Phe Gln Ala Glu Asn Ala 5825 5830 5835 5840 Asn Val Gly Pro Ile Gln Lys Ile Gly Phe Gly Asn Val Glu Ala Met 5845 5850 5855 Glu Ile Asn Leu Asn Ala Asn Leu Phe Lys Ser Phe Leu Tyr Ser Asn 5860 5865 5870 Val Ala Leu Tyr Leu Pro Asp Ala Phe Lys Tyr Thr Pro Glu Asn Ile 5875 5880 5885 Val Ala Pro Ala Asn Ala Asn Thr Tyr Ala Tyr Met Asn Val Arg Leu 5890 5895 5900 Pro Ala Ala Asn Leu Ile Asp Thr Phe Val Asn Ile Gly Ala Arg Trp 5905 5910 5915 5920 Ser Pro Asp Val Met Asp Thr Val Asn Pro Phe Asn His His Arg Asn 5925 5930 5935 Ala Gly Leu Arg Tyr Arg Ser Gln Leu Leu Gly Asn Gly Arg Tyr Cys 5940 5945 5950 Ser Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys Asn Leu 5955 5960 5965 Leu Leu Leu Pro Gly Thr Tyr Thr Tyr Glu Trp Ser Phe Arg Lys Asp 5970 5975 5980 Val Asn Met Ile Leu Gln Ser Ser Leu Gly Asn Asp Leu Arg Val Asp 5985 5990 5995 6000 Gly Ala Thr Ile Asn Ile Gln Ser Ile Asn Leu Tyr Ala Ser Phe Phe 6005 6010 6015 Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala Met Leu Arg Asn 6020 6025 6030 Asp Val Asn Asp Gln Ser Phe Ala Asp Tyr Leu Ser Ser Ala Asn Met 6035 6040 6045 Leu Tyr Pro Ile Pro Ala Asn Thr Thr Asn Leu Pro Ile Ser Ile Pro 6050 6055 6060 Ala Arg Asn Trp Ala Gly Phe Arg Gly Trp Ser Phe Thr Arg Ile Lys 6065 6070 6075 6080 Gln Arg Glu Thr Pro Ala Leu Gly Ser Pro Tyr Asp Pro Tyr Phe Thr 6085 6090 6095 Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Ala Thr Phe Tyr Leu Ser His 6100 6105 6110 Thr Phe Arg Arg Val Ser Ile Met Phe Asp Ser Ser Val Ser Trp Pro 6115 6120 6125 Gly Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys Arg Tyr 6130 6135 6140 Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Ser Asn Met Thr Lys Asp 6145 6150 6155 6160 Trp Phe Met Val Gln Met Leu Ala His Tyr Asn Ile Gly Tyr Gln Gly 6165 6170 6175 Tyr His Leu Pro Glu Ser Tyr Lys Asp Arg Met Tyr Ser Phe Leu Arg 6180 6185 6190 Asn Phe Glu Pro Met Cys Arg Gln Leu Val Asp Val Ala Asn Tyr Ala 6195 6200 6205 Ala Tyr Gln Pro Val Thr Val Gly His Gln His Asn Asn Ser Gly Tyr 6210 6215 6220 Ala Ser Ala Leu Ser Ala Phe Asn Pro Arg Glu Gly His Pro Tyr Pro 6225 6230 6235 6240 Ala Asn Trp Pro Tyr Pro Leu Ile Gly Ala Asn Ala Val Pro Thr Val 6245 6250 6255 Thr Gln Lys Lys Phe Leu Cys Asp Arg Ser Leu Trp Arg Ile Pro Phe 6260 6265 6270 Ser Ser Asn Phe Met Ser Met Gly Thr Leu Thr Asp Leu Gly Gln Asn 6275 6280 6285 Leu Leu Tyr Ser Asn Ser Ala His Ala Leu Asp Met Thr Phe Glu Val 6290 6295 6300 Asp Ala Met Asn Glu Pro Thr Leu Leu Tyr Val Leu Phe Glu Val Phe 6305 6310 6315 6320 Asp Val Ala Arg Val His Gln Pro His Arg Gly Val Ile Glu Val Val 6325 6330 6335 Tyr Leu Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr Met Ala Glu 6340 6345 6350 Gly Gly Ser Ser Glu Glu Glu Leu Arg Ala Ile Val His Asn Leu Gly 6355 6360 6365 Val Ser Pro Phe Phe Leu Gly Thr Phe Asp Lys Arg Phe Pro Gly Phe 6370 6375 6380 Ile Ser Ser Gln Arg Met Ala Cys Ala Ile Val Asn Thr Ala Gly Arg 6385 6390 6395 6400 Glu Thr Gly Gly Val His Trp Leu Ala Met Ala Trp Asn Pro Arg Ser 6405 6410 6415 Lys Thr Phe Tyr Met Phe Asp Pro Phe Gly Phe Ser Asp Thr Lys Leu 6420 6425 6430 Lys Gln Val Tyr Ser Phe Glu Tyr Glu Gly Leu Leu Arg Arg Ser Ala 6435 6440 6445 Ile Ala Ser Ser Pro Asp Arg Cys Val Thr Leu Ala Lys Ser Asn Glu 6450 6455 6460 Thr Ile Gln Gly Pro Asn Ser Ala Ala Cys Gly Leu Phe Cys Cys Met 6465 6470 6475 6480 Phe Leu His Ala Phe Val Asn Trp Pro Asp Asp Pro Phe Asp His Asn 6485 6490 6495 Pro Thr Met Gly Pro Leu Lys Ser Val Pro Asn Tyr Lys Leu Asn Asp 6500 6505 6510 Pro Thr Val Gln Tyr Val Leu Trp Gly Asn Gln Glu Lys Leu Tyr Lys 6515 6520 6525 Phe Leu Glu Lys His Ser Ala Tyr Phe Arg Thr His Ala Ala Ala Ile 6530 6535 6540 Lys Ala Arg Thr Ala Phe Asn Lys Leu Lys Gln Met Ser His Lys Lys 6545 6550 6555 6560 Val Val Ala Ile Ser Glu Ser Ser Ser Asp Glu Glu Val Pro Val Ala 6565 6570 6575 Pro Pro Thr Ala Pro Pro Lys Lys Arg Gln Arg Lys Ala Val Glu Glu 6580 6585 6590 Pro Arg Gly His Gln Ala Met Val Glu Ile Ala Arg Gln Ala Thr Ala 6595 6600 6605 Thr Leu Lys Ala Arg Gly Asp Pro Gly Ser Ile Val Val Gln Thr Phe 6610 6615 6620 Cys Gly Gln Thr Val Asp Thr Asp Lys Glu Gly Asn Val Val Phe Ile 6625 6630 6635 6640 Pro Pro Lys Gln Lys Ser Val Trp Ser Lys Pro Asn Ala Pro Ser Gly 6645 6650 6655 Leu Val Ala Ser Lys Phe Phe Lys Ala Ser Glu His Lys Trp Gln Ser 6660 6665 6670 Ala Met Glu Leu Ala Leu Lys Val Leu Thr Ala Tyr Gln Val Asp His 6675 6680 6685 Ser Lys Leu Thr Leu Leu Pro Asp Glu Gly Thr Leu Glu Cys Phe Lys 6690 6695 6700 Lys Ala Val Gln Ala Tyr Ile Thr Thr Ser Lys Thr His Val Thr Tyr 6705 6710 6715 6720 Thr Phe Thr Asn Gln Lys Thr Phe Leu His Val Ala Gly Arg Leu Leu 6725 6730 6735 Leu Asp Phe Val Ile Lys Ala Ala Glu Leu Ala Pro Gly Val Asn Pro 6740 6745 6750 Ser Gly Cys Val Val Trp Gln His Gly Cys Gln Ser Ser Leu Met Cys 6755 6760 6765 Leu His Gly Ser Pro Met Ile Gln Lys Glu Gln Leu Val Glu Met Asp 6770 6775 6780 Val Asn Ser Glu Asn Ala Gln Arg Ala Leu Lys Glu Asn Pro Glu Lys 6785 6790 6795 6800 Thr Lys Ile Val Ser Asn Arg Trp Gly Arg Asn Val Val Gln Phe Lys 6805 6810 6815 Asn Glu Asp Ala Phe Cys Cys Ser Met Asp Val Asn Met Ser Gly Gly 6820 6825 6830 Asn Phe Ser Gly Ala Ser Cys Gly Met Phe Tyr Thr Asp Gly Pro Lys 6835 6840 6845 Ala Ile Met Ala Phe Gln Gln Ile Met Ala Phe Leu Lys Ala Cys Tyr 6850 6855 6860 Pro Ser Met Pro Asn Ala Glu Ser His Leu Leu Met Pro Leu Lys Cys 6865 6870 6875 6880 Glu Cys Asn Trp Asn Ser Asn Leu Pro Leu Leu Gly Arg Gln Thr Cys 6885 6890 6895 Lys Ile Thr Pro Phe Ser Leu Ala Ser Ala Ser His Ile Asp Lys Ser 6900 6905 6910 Glu Val Asp Asp Gln Lys Met Leu Ala Thr Leu Asn Asn Pro Ala Met 6915 6920 6925 Leu Val Phe Gln Cys Cys Asn Pro Val Tyr Arg Asn Ser Lys Ala Ala 6930 6935 6940 Pro Gln Lys Asn Cys Asp Phe Lys Ile Ser Val Asp Leu Val Ser 6945 6950 6955 6960 Cys Val Gln Ile Ala Lys Gln Ile Trp Thr Ser Thr Val Gly Glu Arg 6965 6970 6975 Pro Pro Ile Lys Phe Pro Glu Phe His Trp Ser Asp Glu His Arg Tyr 6980 6985 6990 Gln Asn Thr Ile Leu Pro Gln Gly Gln His Asp Asp Asp Leu Val Leu 6995 7000 7005 Phe Met Ser Glu Glu Pro Val Ser Gly Thr Thr Val Glu Thr Lys Glu 7010 7015 7020 Asp Ile His Thr Pro Pro Asn Ser Pro Val Leu Glu Thr Ile Ser Leu 7025 7030 7035 7040 Ser Pro Glu Pro Glu Ala Glu Ala Cys Pro Asn Thr Asp Lys Tyr Leu 7045 7050 7055 Ser Thr Asn Leu Leu Cys Lys His Leu Gln Arg Gln Ser Ala Ile Val 7060 7065 7070 Leu Asp Ser Ile Lys Asp His Leu Gln Val Pro Thr Ser Val Ser Glu 7075 7080 7085 Leu Ser Cys Ala Tyr Glu Arg Ser Leu Phe Ser Pro Asn Thr Pro Pro 7090 7095 7100 Arg Gln Gln Ser Asn Gly Thr Cys Glu Ala Asn Pro Lys Leu Asn Phe 7105 7110 7115 7120 Tyr Pro Thr Phe Leu Val Pro Glu Thr Leu Ala Thr Tyr His Ile Phe 7125 7130 7135 Phe Val Asn Gln Lys Ile Pro Val Ser Cys Arg Ala Asn Arg Ala Lys 7140 7145 7150 Ala Asp Arg Ala Leu Ala Leu Gln Glu Gly Asp Cys Leu Pro Asp Tyr 7155 7160 7165 Glu Thr Met Asp Thr Val Ser Arg Val Phe Glu Gly Leu Gly Gly Glu 7170 7175 7180 Val Val Ala Asp His Ala Leu Gln Asn Asn Asp Ser Val Leu Val Glu 7185 7190 7195 7200 Leu Lys Glu Asp Asn Pro Arg Leu Ala Val Val Lys Arg Asn Leu Ser 7205 7210 7215 Val Ser His Phe Ala Tyr Pro Ala Val His Leu Pro Pro Lys Val Ile 7220 7225 7230 Thr Thr Val Met Asp Asn Leu Leu Val Lys Arg Ser Thr Pro Ser Ala 7235 7240 7245 Asp Val Ser Glu Leu Asp Pro Glu Gly Gly Gin Glu Val Val Ser Asp 7250 7255 7260 Thr Glu Leu Ser Lys Trp Leu Asn Thr Ser Asp Pro Glu Ala Leu Glu 7265 7270 7275 7280 Lys Gln Arg Lys Thr Val Met Gly Ser Val Leu Val Thr Val Val Leu 7285 7290 7295 Glu Cys Met Gln Arg Leu Phe Thr Ser Glu Glu Met Val Lys Lys Ile 7300 7305 7310 Gly Glu Ala Leu His Tyr Thr Phe Arg His Gly Tyr Val Ser Leu Ala 7315 7320 7325 Cys Lys Ile Ser Asn Val Glu Leu Thr Asn Val Val Thr Tyr Met Gly 7330 7335 7340 Ile Leu His Glu Asn Arg Leu Gly Gin Thr Thr Leu His His Thr Val 7345 7350 7355 7360 Gln Gly Glu Ala Arg Arg Asp Tyr Ile Arg Asp Ser Ile Phe Leu Ile 7365 7370 7375 Leu Ile His Ala Trp Gln Thr Ala Met Gly Ile Trp Gln Gln Cys Leu 7380 7385 7390 Glu Glu Glu Asn Leu Lys Glu Leu Ala Lys Leu Leu Gln Arg Ile Lys 7395 7400 7405 Lys Pro Leu Tyr Thr Glu Thr Ser Gln Arg Leu Met Gly Lys Leu Leu 7410 7415 7420 Ala Asn Val Val Phe Pro Lys Leu Leu Gln Thr Phe His Lys Gly 7425 7430 7435 7440 Leu Pro Asp Ile Val Ser Gln Ser Met Met Gln Asn Phe Arg Ser Phe 7445 7450 7455 Ile Leu Glu Arg Ser Gly Ile Leu Pro Ser Met Thr Cys Ala Leu Pro 7460 7465 7470 Thr Asp Phe Val Pro Ile His Phe Lys Glu Cys Pro Pro Thr Met Trp 7475 7480 7485 Pro Tyr Thr Tyr Leu Leu Lys Leu Ala Asn Phe Phe Met Tyr His Asn 7490 7495 7500 Asp Leu Cys Tyr Asp Val Gly Gly Glu Gly Leu Leu Glu His Tyr Cys 7505 7510 7515 7520 Arg Cys Asn Leu Cys Thr Pro His Arg Cys Leu Ala Thr Asn Pro Ala 7525 7530 7535 Met Leu Asn Glu Thr Gln Leu Ile Gly Thr Phe Asp Ile Arg Gly Pro 7540 7545 7550 Gly Gly Glu Asn Gly Ala Glu Ser Ser Ser Gly Leu Lys Leu Thr Ala 7555 7560 7565 Gly Met Trp Thr Ser Ala Phe Leu Arg Lys Phe Glu Ser Ser Asp Tyr 7570 7575 7580 His Ala His Lys Ile Gln Phe Tyr Glu Asn Gln Ser Lys Pro Pro Ser 7585 7590 7595 7600 Val Glu Pro Thr Pro Cys Val Ile Thr Gln Ser Ser Ile Leu Ala Gln 7605 7610 7615 Leu His Asp Ile Lys Lys Ala Arg Glu Glu Phe Leu Leu Lys Ser Gly 7620 7625 7630 Gln Gly Gln Tyr Leu Asp Pro Gln Thr Gly Glu Pro Leu Asn Ala Ala 7635 7640 7645 Asp Pro Ser Val Glu Ser Ser His Glu Phe Gln Gly Asp Gly Arg His 7650 7655 7660 Arg Asn Pro Lys Arg Gly Arg Asn Phe Arg Gln Arg Gly Gly Pro Arg 7665 7670 7675 7680 Lys Pro Pro Arg Val His Ala Gly Ala Glu Arg Asp Ala Arg Gly Thr 7685 7690 7695 Ser Ser Met Ser Ser Lys Glu Met Glu Asp Thr Glu Thr Leu Ser Val 7700 7705 7710 Glu Glu Ile Ser Gly Ser Glu Glu Asp Leu Glu Ser Leu Pro Glu Ser 7715 7720 7725 Met Pro Glu Gln Ser Glu Met Pro Glu Glu Arg Ala Pro Arg Trp Asp 7730 7735 7740 Gln Lys Glu Lys Pro Leu Ala Lys Gln Lys Leu Lys Arg Arg Pro Thr 7745 7750 7755 7760 Pro Leu Lys Ser Pro Lys Pro Ala Ala Gly Glu Asp Asn His Gln Thr 7765 7770 7775 Arg Gln Leu Arg Asn Arg Ile Phe Pro Thr Leu Tyr Ala Ile Phe Gln 7780 7785 7790 Gln Ser Arg Gly Ser Pro Thr Ala Leu Lys Ile Lys Asn Arg Ser Leu 7795 7800 7805 Arg Ser Leu Leu Arg Ser Cys Leu Tyr His Lys Ser Glu Glu Gln Leu 7810 7815 7820 Leu Arg Thr Arg Asn Asp Ala Glu Ala Leu Leu Asn Lys Tyr Cys Gln 7825 7830 7835 7840 Gly Leu Glu Ser Gly Thr Asp Met Ser Ser Lys Glu Met Glu Asp Thr 7845 7850 7855 Glu Thr Leu Ser Val Glu Glu Ile Ser Gly Ser Glu Glu Asp Leu Glu 7860 7865 7870 Ser Leu Pro Glu Ser Met Pro Glu Gln Ser Glu Met Pro Glu Glu Arg 7875 7880 7885 Ala Pro Arg Trp Asp Gln Lys Glu Lys Pro Leu Gly Lys Gln Pro Arg 7890 7895 7900 Asn Tyr His Ser Trp Arg Ala His Lys Phe Lys Ile Leu Ser Cys Leu 7905 7910 7915 7920 Gly Ala Ser Gly Asn Asn Val Ala Phe Thr Arg Arg Tyr Met Leu Phe 7925 7930 7935 Arg Glu Gly Val Asn Leu Pro Asn Asn Ile Ile His Tyr Tyr Asn Ser 7940 7945 7950 Arg Tyr Arg Ser Lys Thr Glu Ala Gln Ala Gln Thr Asp Pro Thr Lys 7955 7960 7965 Glu Pro Lys Ala Ser Arg Arg Met Ser Lys Glu Ile Pro Thr Pro Tyr 7970 7975 7980 Met Trp Ser Tyr Gln Pro Gln Thr Gly His Ala Ala Gly Ala Ser Gln 7985 7990 7995 8000 Asp Tyr Ser Thr Gln Met Asn Trp Phe Ser Ala Gly Pro Ser Met Ile 8005 8010 8015 Ser Gln Val Tyr Gly Ile Arg Asp Leu Arg Asn Lys Val Leu Ile Thr 8020 8025 8030 Gln Ala Glu Ile Thr Lys Thr Pro Arg Thr Ile Met Asp Pro Pro Ile 8035 8040 8045 Trp Pro Ala Ala Met Leu Val Gln Glu Ala Ala Pro Lys Thr Val 8050 8055 8060 Thr Leu Pro Arg Asn His Thr Leu Glu Gln Ala Met Thr Asn Ser Gly 8065 8070 8075 8080 Ala Gln Leu Ala Gly Gly Arg Gln Leu Cys Pro Ser Gln Ile Gly Ile 8085 8090 8095 Lys Ser Pro Val Leu Ala Gly Thr Gly Ile Gln Leu Ser Glu Asp Ile 8100 8105 8110 Pro Ser Ala Ser Trp Ile Arg Pro Asp Gly Ile Phe Gln Leu Gly Gly 8115 8120 8125 Gly Ser Arg Ser Ser Phe Ser Pro Thr Gln Ala Phe Leu Thr Leu Gln 8130 8135 8140 Gln Ala Ser Ser Thr Pro Arg Ala Gly Gly Val Gly Thr Tyr Gln Phe 8145 8150 8155 8160 Val Arg Glu Phe Val Pro Glu Val Tyr Leu Asn Pro Phe Ser Gly Pro 8165 8170 8175 Pro Asp Thr Phe Pro Asp Gln Phe Ile Pro Asn Tyr Asp Ile Val Thr 8180 8185 8190 Asn Ser Val Asp Gly Tyr Asp Met Ala Met Thr Lys Glu Ser Met Asp 8195 8200 8205 Gln Val Glu Val Asn Cys Leu Cys Ala Gln His Gly Gln Thr Cys Thr 8210 8215 8220 Arg Pro Arg Cys Phe Ala Lys Glu Gly Leu Cys Ala Asn Trp Phe Tyr 8225 8230 8235 8240 Asn Pro Ala Leu Ala Phe Glu Gly Phe Asp Ile Pro Asp Ser Tyr Gln 8245 8250 8255 Glu Gly His Gly Val Asp Ile Glu Val Lys Cys Ser His His Ser Ser 8260 8265 8270 Lys Leu Cys His Asn Gly His Asp Met Ile Cys Ser Tyr Ser Arg Leu 8275 8280 8285 Gly Ser His Ile Asn Ile Arg Cys Ile Cys Asn Lys Pro Arg Pro His 8290 8295 8300 Met Ser Leu Ile Glu Ala Ala Cys Ser Met Tyr Asn Leu Asn Met Arg 8305 8310 8315 8320 Phe Ser Phe Phe Ile Ala Ala Val Leu Phe Cys Thr Thr Gly Ala Ser 8325 8330 8335 Asn Asp Ile Val Thr Cys Cys Ala His Thr Pro Cys Leu Leu His Leu 8340 8345 8350 Glu Val Gly Leu Gly Ala Asn Val Ser Trp Ile Asn Ser Asp Thr Gly 8355 8360 8365 Gln Ala Pro Leu Cys Leu Ser Asn Gly Met Cys Asn Ala Thr Gln Gln 8370 8375 8380 Gly Leu Gln Phe Ser Ala Asn Phe Ser Glu Asp Gly Leu Tyr Ile Ala 8385 8390 8395 8400 Leu Ile Lys Glu Ser Asn Tyr Glu Gly Ala Glu His Tyr Tyr Leu Val 8405 8410 8415 Tyr Ile Tyr Gly Asp Cys Tyr Gln Thr Ala Asn Glu Ser Ala His Gly 8420 8425 8430 Pro Ile Ser Arg Pro Leu Asn Glu Met Pro Leu His Ser Val Thr Ile 8435 8440 8445 Asn Ala Ser Leu Phe Tyr Pro Ala Phe Leu Glu Leu Pro Pro Gln Tyr 8450 8455 8460 Ser Asn Asp Leu Ser Asn Val Arg Trp Tyr Lys Val Asp Pro Ser Gly 8465 8470 8475 8480 Phe Gln Ala Gln Lys Ile Ser Lys Val Arg Ser Gly Gly Arg Lys Glu 8485 8490 8495 Asn Leu His Pro Asn Trp Ala Leu Val Thr Tyr Thr Gly Asp Leu Leu 8500 8505 8510 Val Leu His Val Ser Pro Asn Thr Leu Gly Leu Trp Leu Ala Ala Val 8515 8520 8525 Gln His Arg Gly Gly Arg Thr Asn Phe Ile Thr Phe Asn Ile Thr Val 8530 8535 8540 Pro Asn Trp Gln Gln Asn Leu Val Ala Ile Phe Asn Gln His Glu Pro 8545 8550 8555 8560 Pro Lys Met Gly Asn Asn Tyr Glu Asp Ser Phe Met Glu Trp Thr Leu 8565 8570 8575 Phe Lys Lys Leu Lys Lys Gly Leu Phe Arg Val Thr Cys Arg Ala Lys 8580 8585 8590 Ser Ile Phe Pro Glu Cys Thr Leu Asn Ile Thr Arg Asp Gly Thr Phe 8595 8600 8605 Leu Leu Ile Gly Glu Ser Lys Lys Thr Pro Tyr Val Ile Leu Leu Pro 8610 8615 8620 Phe Phe Ala Asn Pro Lys Glu Asp Thr Pro Ile Leu Met Ala Leu Ser 8625 8630 8635 8640 His Ser Val Pro Val Ala Ile Pro Asn Thr Ala Met Pro Val Tyr Ile 8645 8650 8655 Ser Ile Met Phe Phe Ile Val Ala Met Leu Ala Thr Leu Ser Leu Leu 8660 8665 8670 Met Gly Leu Asn Asn Lys Ile Arg Ser Met Met Ala Leu Val Leu Val 8675 8680 8685 Asn Gly Lys Leu Leu Gly Glu Thr Thr Leu Ser Phe Lys His Ile Arg 8690 8695 8700 Lys Leu Ala Arg Thr His Lys Trp Lys Tyr Gln Ser Trp Glu Glu Gly 8705 8710 8715 8720 Thr Leu Val Asp Ile Glu Thr Gln Asp Pro Asp Val Leu Ala Lys Ile 8725 8730 8735 Met Lys Arg Thr Arg Arg Ala Leu Pro Ala Asn Tyr Asp Pro Val Tyr 8740 8745 8750 Pro Tyr Asp Ala Pro Gly Ser Ser Thr Gln Pro Phe Phe Asn Asn 8755 8760 8765 Lys Gln Gly Leu Thr Glu Ser Pro Gly Thr Leu Ala Val Asn Val 8770 8775 8780 Ser Pro Pro Leu Thr Phe Ser Thr Leu Gly Ala Ile Lys Leu Ser Thr 8785 8790 8795 8800 Gly Pro Gly Leu Thr Leu Asn Glu Gly Lys Leu Gln Ala Ser Leu Gly 8805 8810 8815 Pro Gly Leu Ile Thr Asn Thr Glu Gly Gin Ile Thr Val Glu Asn Val 8820 8825 8830 Asn Lys Val Leu Ser Phe Thr Ser Pro Leu His Lys Thr Glu Asn Thr 8835 8840 8845 Val Ser Leu Ala Leu Gly Asp Gly Leu Glu Asp Glu Asn Gly Thr Leu 8850 8855 8860 Lys Val Thr Phe Pro Thr Pro Pro Pro Leu Gln Phe Ser Pro Pro 8865 8870 8875 8880 Leu Thr Glu Thr Gly Gly Thr Ile Ser Leu Pro Leu Gln Asp Ser Met 8885 8890 8895 Gln Val Thr Asn Gly Lys Leu Gly Val Lys Pro Thr Thr Tyr Ala Pro 8900 8905 8910 Pro Leu Lys Lys Thr Asp Gln Gln Val Ser Leu Gln Val Gly Ser Gly 8915 8920 8925 Leu Thr Val Ile Asn Glu Gln Leu Gln Ala Val Gln Pro Pro Ala Thr 8930 8935 8940 Thr Tyr Asn Glu Pro Leu Ser Lys Thr Asp Asn Ser Val Ser Leu Gln 8945 8950 8955 8960 Val Gly Ala Gly Leu Ala Val Gln Ser Gly Ala Leu Val Ala Thr Pro 8965 8970 8975 Pro Pro Leu Thr Phe Thr Ser Pro Leu Glu Lys Asn Glu Asn Thr 8980 8985 8990 Val Ser Leu Gln Val Gly Ala Gly Leu Ser Val Gln Asn Asn Ala Leu 8995 9000 9005 Val Ala Thr Pro Pro Pro Leu Thr Phe Ala Tyr Pro Leu Val Lys 9010 9015 9020 Asn Asp Asn His Val Ala Leu Ser Ala Gly Ser Gly Leu Arg Ile Ser 9025 9030 9035 9040 Gly Gly Ser Leu Thr Val Ala Thr Gly Pro Gly Leu Ser His Gln Asn 9045 9050 9055 Gly Thr Ile Gly Ala Val Val Gly Ala Gly Leu Lys Phe Glu Asn Asn 9060 9065 9070 Ala Ile Leu Ala Lys Leu Gly Asn Gly Leu Thr Ile Arg Asp Gly Ala 9075 9080 9085 Ile Glu Ala Thr Gln Pro Pro Ala Ala Pro Ile Thr Leu Trp Thr Gly 9090 9095 9100 Pro Asp Pro Ser Ile Asn Gly Phe Ile Asn Asp Thr Pro Val Ile Arg 9105 9110 9115 9120 Cys Phe Ile Cys Leu Thr Arg Asp Ser Asn Leu Val Thr Val Asn Ala 9125 9130 9135 Ser Phe Val Gly Glu Gly Gly Tyr Arg Ile Val Ser Pro Thr Gln Ser 9140 9145 9150 Gln Phe Ser Leu Ile Met Glu Phe Asp Gln Phe Gly Gln Leu Met Ser 9155 9160 9165 Thr Gly Asn Ile Asn Ser Thr Thr Thr Trp Gly Glu Lys Pro Trp Gly 9170 9175 9180 Asn Asn Thr Val Gln Pro Arg Pro Ser His Thr Trp Lys Leu Cys Met 9185 9190 9195 9200 Pro Asn Arg Glu Val Tyr Ser Thr Pro Ala Ala Thr Ile Ser Arg Cys 9205 9210 9215 Gly Leu Asp Ser Ile Ala Val Asp Gly Ala Pro Ser Arg Ser Ile Asp 9220 9225 9230 Cys Met Leu Ile Ile Asn Lys Pro Lys Gly Val Ala Thr Tyr Thr Leu 9235 9240 9245 Thr Phe Arg Phe Leu Asn Phe Asn Arg Leu Ser Gly Gly Thr Leu Phe 9250 9255 9260 Lys Thr Asp Val Leu Thr Phe Thr Tyr Val Gly Glu Asn Gln Met Glu 9265 9270 9275 9280 Gly Asn Cys Thr Ala Glu Thr Thr Cys His Val Thr Ala Val Ile Glu 9285 9290 9295 Pro Thr Val Leu Asp Tyr Ser Cys Ile Pro Val Asp Gly Phe Val Glu 9300 9305 9310 Ile Thr Asp Pro Cys Phe Ala Thr Gln Glu Asn Val Tyr Tyr Tyr Ser 9315 9320 9325 Pro Asn Glu Leu Leu Thr Asn Thr Tyr Val Leu Cys Cys Cys Glu Arg 9330 9335 9340 Thr Val Tyr Lys Val Thr Phe Pro Glu Gly Gly Ser Leu Thr Ala Arg 9345 9350 9355 9360 Val Tyr Glu Lys Met Glu Gly Asn Cys Thr Ala Glu Thr Thr Cys His 9365 9370 9375 Val Thr Ala Val Val Arg Val Pro Lys Tyr Cys Asn Cys Phe Ala Leu 9380 9385 9390 Cys Tyr Glu Ile Pro Ile Leu Trp Asp Asp Val Leu His Arg His Glu 9395 9400 9405 Lys Leu Leu Phe Gly Gly Phe Thr Cys Asn Ala Gly Ile Glu Leu Ile 9410 9415 9420 Val Thr Lys His Cys Cys Leu Ala Glu Ala Gln Thr Trp Gln Leu His 9425 9430 9435 9440 Cys His Cys Ser Asn Ser Leu Ser Leu Gln Cys Met Ala Ser Lys His 9445 9450 9455 Val Val Gln Lys Val Ile Glu Asp Phe Val Lys Gly Gly Ala Met Asn 9460 9465 9470 Lys Lys Tyr Met Trp Tyr Arg Glu Phe Val Asn Ser Ser Arg Pro Asp 9475 9480 9485 Glu Ile Asn Tyr Val Gly Ser Ile Ile Phe Arg Asn Thr His Tyr Ile 9490 9495 9500 Tyr Phe Arg Leu Ser Phe Phe Arg Thr Val His Lys Ala Cys Met Glu 9505 9510 9515 9520 Ala Ile Lys Arg Cys Ile Asn Pro Glu Leu Gly Val Val Leu Lys Ser 9525 9530 9535 Thr Tyr Asn Tyr Trp Leu Val Leu Lys Cys Lys Ser Cys Ser Leu Gln 9540 9545 9550 Asn Tyr Cys Ser Leu Lys Asn Cys Ala Val Trp Val Arg Ser Val Ile 9555 9560 9565 Glu Arg Val Leu Lys Glu Val Glu Lys Thr Pro Val Val Leu His Gln 9570 9575 9580 Thr Thr Ser Lys Ala Glu Glu Arg Arg Gln Ser Ala Leu Arg Gln Ala 9585 9590 9595 9600 Met Met Tyr Gly Arg Cys Arg Gln Ile Arg Asn Leu Cys Leu Val Asn 9605 9610 9615 Leu Asn Ala Phe Leu His Phe Met Ala His His Arg Leu Pro Arg Val 9620 9625 9630 Cys Val Lys Gly Ile Ile His Phe Glu Glu Asp Phe Val Arg Glu Leu 9635 9640 9645 Gly Ser Met Leu Glu Ser Pro Met Glu Phe Leu Phe Asp Thr Ile Asp 9650 9655 9660 Asp Val Thr Ala Ser Ile Phe Cys Glu Ser Met Phe Lys Ala Val Asp 9665 9670 9675 9680 Lys Asn Lys Pro Gly Ile Thr Phe Lys Val Val Phe Tyr Ser Gln Leu 9685 9690 9695 Gly Phe Glu Tyr Asp Asp Ala Leu Ala His Phe Lys Gly Thr Leu Ile 9700 9705 9710 Lys Glu Ile Ser Asp Val Val Asn Asn His Pro Asn Val Asn Asn Ala 9715 9720 9725 Phe Arg Gly Arg Glu Ile Val Thr Val Ser Leu Leu Glu Val Phe Ser 9730 9735 9740 Phe Cys Ser Met Ala Thr Ala Asp Ser Glu Pro Arg Leu Lys Cys Leu 9745 9750 9755 9760 Cys Phe Thr Ile Thr Val Glu Pro Ala Leu Ala Glu Ile Leu Leu Ser 9765 9770 9775 Leu Asp Lys Asn Ile Gly Ala Val Met Cys Ala Gly Leu Lys Ala Phe 9780 9785 9790 Leu Lys Arg Trp Ile Val Thr Ala Cys Val Gln Glu Leu Pro Val Lys 9795 9800 9805 Glu Gly Met Glu Val Phe Val Ala Ile Ser Trp His Tyr Val Ala Cys 9810 9815 9820 Thr Thr Gln Leu Gln Asn Phe Leu Thr Glu Gly Met Thr Asn Tyr Leu 9825 9830 9835 9840 Asp Ser Phe Phe Lys Asp Phe Gly Ile Thr His Gly His Asp Phe Thr 9845 9850 9855 Thr Ala Glu Leu Ala Leu Phe Thr Val Lys Cys Val Ser Gly Met Leu 9860 9865 9870 Ser Gly Val Leu Leu Phe Cys Arg Met Ala Ser Gly Ala Ala Tyr Phe 9875 9880 9885 Tyr Glu Cys Asn Ile Thr Leu Lys Asp His Leu Val Ala Ile Ile Arg 9890 9895 9900 Asp Gln Asp Arg Glu Thr Asp Leu Cys Arg Gly Val Ser Met Leu Leu 9905 9910 9915 9920 Gln Ser Ile Phe Asn Leu Arg Ala Val Arg Arg Leu Ser Ser His Ser 9925 9930 9935 Gly Pro Leu Ile Val Thr Ser Ile Val Tyr Thr Ser Pro Asn Ser Met 9940 9945 9950 Cys Thr Leu Leu Lys Glu Glu Val Glu Ser Leu Ile Lys Val Tyr Ile 9955 9960 9965 Arg Gly Trp Ile Glu Gly Leu Gly Cys Gly Ile Asp Glu Asp Leu His 9970 9975 9980 Met Gln Trp Leu Asn Glu Val Lys Val His Phe Phe Ser Cys Met Ala 9985 9990 9995 10000 Ala Leu Gly Val Ser Met Gly Ala Cys Phe Cys Leu Arg Leu His Lys 10005 10010 10015 Ser Leu Val Glu Ser Val Cys Ala Gln Leu Glu Leu Thr Asp Phe Leu 10020 10025 10030 Pro Ser Glu Leu Arg Arg Val Val Phe Ser Leu Ile Arg Ala Gln Glu 10035 10040 10045 Val Arg Pro Cys Pro Thr Phe Ala Ala Ala Val Ser Val Leu Ser Gly 10050 10055 10060 Asp Tyr Leu Glu Phe Tyr Leu Phe Ile Gly Ser Asp Thr Glu Gly Phe 10065 10070 10075 10080 Gly Ser Val Ala Asp Ile Gln Ala Thr Ala Val Ser Leu Gln Glu Ala 10085 10090 10095 Leu Arg Arg Asp Phe Cys Gln Leu Ile Pro Glu Asp Cys Ala His Leu 10100 10105 10110 Ala Leu Gly Leu Ser Val Thr Trp Glu Pro Asn Ile Leu Val Thr Ala 10115 10120 10125 Glu <210> 4 <211> 170 <212> PRT <213> Artificial Sequence <220> <223> APQA1701_E1B19K <400> 4 Met Asp Pro Leu Lys Ile Cys Glu Asn Tyr Leu Thr Phe Arg Ala Ile 1 5 10 15 Ile Arg Gly Ser Thr Leu Ser Pro Gly Phe Phe Arg Arg Trp Cys Phe 20 25 30 Pro Ala Leu Ala Asp Val Val Gly Asn Ile Val Glu Gln Glu Glu Gly 35 40 45 Arg Phe Trp Gln Ile Leu Pro Glu Asn His Thr Phe Trp Gly Leu Leu 50 55 60 Arg Arg Gly Phe Thr Val Ala Ser Phe Thr Glu Ile Ile Thr Ala Ala 65 70 75 80 Gln Leu Glu Asn Arg Gly Arg Gln Leu Ala Phe Leu Ala Phe Ile Ser 85 90 95 Phe Leu Leu Arg Asn Trp Pro Ser Asp Ser Val Val Pro Glu Ala Asp 100 105 110 Arg Leu Asp Leu Val Cys Ala Pro Ala Trp Ser Arg Met Gln Ile Trp 115 120 125 Ser Gln Thr Ala Arg Leu Ile Asn Asp Leu Gln Asp Ser Val Leu Glu 130 135 140 Glu Gln Gly Ser Ala Glu Glu Glu Glu Cys Glu Glu Ala Leu Leu Ala 145 150 155 160 Gly Asp Ser Asp Asp Pro Leu Phe Gly *** 165 170 <210> 5 <211> 164 <212> PRT <213> Artificial Sequence <220> <223> APQA1701_40P_E1B19K <400> 5 Met Asp Pro Leu Lys Ile Cys Glu Asn Tyr Leu Thr Phe Arg Ala Ile 1 5 10 15 Ile Arg Gly Ser Thr Leu Ser Pro Gly Phe Phe Arg Arg Trp Cys Phe 20 25 30 Pro Ala Leu Ala Asp Val Val Gly Asn Ile Val Glu Gln Glu Glu Gly 35 40 45 Arg Phe Trp Gln Ile Leu Pro Glu Asn His Thr Phe Trp Gly Leu Leu 50 55 60 Arg Arg Gly Phe Thr Val Ala Ser Phe Thr Glu Ile Ile Thr Ala Ala 65 70 75 80 Gln Leu Glu Asn Arg Gly Arg Gln Leu Ala Phe Leu Ala Phe Ile Ser 85 90 95 Phe Leu Leu Arg Asn Trp Pro Ser Asp Ser Val Val Pro Glu Ala Asp 100 105 110 Arg Leu Asp Leu Val Cys Ala Pro Ala Trp Ser Gln Thr Ala Arg Leu 115 120 125 Ile Asn Asp Leu Gln Asp Ser Val Leu Glu Glu Gln Gly Ser Ala Glu 130 135 140 Glu Glu Glu Cys Glu Glu Ala Leu Leu Ala Gly Asp Ser Asp Asp Pro 145 150 155 160 Leu Phe Gly *** <210> 6 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> APQA1701_E3_12.5K <400> 6 Met Ala Met Thr Lys Glu Ser Met Asp Gln Val Glu Val Asn Cys Leu 1 5 10 15 Cys Ala Gln His Ala Gln Thr Cys Thr Arg Pro Arg Cys Phe Ala Lys 20 25 30 Glu Gly Leu Cys Ala Asn Trp Phe Tyr Asn Pro Ala Leu Ala Phe Glu 35 40 45 Gly Phe Asp Ile Pro Asp Ser Tyr Gln Glu Gly His Gly Val Asp Ile 50 55 60 Glu Val Lys Cys Ser His His Ser Ser Lys Leu Cys His Asn Gly His 65 70 75 80 Asp Met Ile Cys Ser Tyr Ser Arg Leu Gly Ser His Ile Asn Ile Arg 85 90 95 Cys Ile Cys Asn Lys Pro Arg Pro His Met Ser Leu Ile Glu Ala Ala 100 105 110 Cys Ser Met Tyr Asn Leu Asn *** 115 120 <210> 7 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> APQA1701_40P_E3_12.5K <400> 7 Met Ala Met Thr Lys Glu Ser Met Asp Gln Val Glu Val Asn Cys Leu 1 5 10 15 Cys Ala Gln His Gly Gln Thr Cys Thr Arg Pro Arg Cys Phe Ala Lys 20 25 30 Glu Gly Leu Cys Ala Asn Trp Phe Tyr Asn Pro Ala Leu Ala Phe Glu 35 40 45 Gly Phe Asp Ile Pro Asp Ser Tyr Gln Glu Gly His Gly Val Asp Ile 50 55 60 Glu Val Lys Cys Ser His His Ser Ser Lys Leu Cys His Asn Gly His 65 70 75 80 Asp Met Ile Cys Ser Tyr Ser Arg Leu Gly Ser His Ile Asn Ile Arg 85 90 95 Cys Ile Cys Asn Lys Pro Arg Pro His Met Ser Leu Ile Glu Ala Ala 100 105 110 Cys Ser Met Tyr Asn Leu Asn *** 115 120

Claims (10)

Novel Canine Adenovirus Type 2 strain deposited with accession number KCTC13956BP.
According to claim 1,
The canine adenovirus type 2 strain entrusted with the accession number KCTC13956BP is characterized in that it is a strain consisting of the nucleotide sequence of SEQ ID NO: 2,
Canine Adenovirus Type 2 strain deposited with accession number KCTC13956BP.
According to claim 1,
The E1b 19K protein of the Canine Adenovirus Type 2 strain deposited with the accession number KCTC13956BP consists of the amino acid sequence of SEQ ID NO: 5; E3 12.5K protein is characterized in that consisting of the amino acid sequence of SEQ ID NO: 7,
Canine Adenovirus Type 2 strain deposited with accession number KCTC13956BP.
A method for preparing an antigen for enzymatic immunization, comprising the steps of:
(a) inoculating the cultured cells with canine adenovirus type 2 (APQA1701-40P) deposited with accession number KCTC13956BP and culturing; and
(b) purifying the cultured canine adenovirus type 2 (APQA1701-40P).
5. The method of claim 4,
The step (a) comprises the steps of (a-1) inoculating and proliferating canine adenovirus type 2 in cultured cells and then culturing; And (a-2) characterized in that it comprises the step of centrifuging the culture,
A method for preparing an antigen for use in enzymatic immunization.
According to claim 4, wherein the cultured cells, MDCK (Madin-Darby canine kidney) cells, dog primary cultured cells (primary dog kidney cells), characterized in that the animal cells, or a combination thereof,
A method for preparing an antigen for use in enzymatic immunization.
An antigen for enzymatic immunization prepared by the method according to any one of claims 4 to 6.
An enzyme immunization technique kit for detecting an antibody against canine adenovirus type 2 comprising the antigen according to claim 7.
A method for detecting antibodies to canine adenovirus type 2 comprising the steps of:
(a) contacting the biological sample with the antigen for the enzymatic immunization technique according to claim 7; and
(b) detecting the presence of the antibody bound to the antigen for the enzyme immunization technique in the sample by the enzyme immunization technique.
10. The method of claim 9, wherein the biological sample is characterized in that at least one selected from the group consisting of plasma, whole blood, serum, urine and saliva,
A method for detecting an antibody against canine adenovirus type 2.

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