阅读说明：本技术 新型嗜水气单胞菌噬菌体Aer-HYP-3及其在抑制嗜水气单胞菌细菌生长方面的用途 (Novel aeromonas hydrophila phage Aer-HYP-3 and application thereof in inhibiting growth of aeromonas hydrophila bacteria ) 是由 尹圣君 俊苏云 权安升 蒋基英 康桑贤 于 2019-02-18 设计创作，主要内容包括：本发明涉及一种从自然界分离出的肌尾噬菌体科噬菌体Aer-HYP-3(登录号：KCTC 13479BP),其具有杀死嗜水气单胞菌细菌的能力,并且具有由SEQ ID NO:1表示的基因组,还涉及一种使用包含所述噬菌体作为活性成分的组合物来预防和治疗由嗜水气单胞菌引起的疾病的方法。(The present invention relates to a bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) of Myoviridae, which is isolated from the natural world, has the ability to kill Aeromonas hydrophila bacteria, and has a genome represented by SEQ ID NO:1, and a method for preventing and treating diseases caused by Aeromonas hydrophila using a composition comprising the bacteriophage as an active ingredient.)

1. A bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) of Myoviridae having an ability to kill Aeromonas hydrophila and having a genome represented by SEQ ID NO: 1.

2. A composition for preventing and treating diseases caused by aeromonas hydrophila, comprising the bacteriophage Aer-HYP-3 (accession No. KCTC 13479BP) according to claim 1 as an active ingredient.

3. The composition according to claim 2, wherein the composition is used in the form of a medicated bath agent or a feed additive.

4. A method for preventing and treating a disease caused by aeromonas hydrophila, comprising:

an animal other than a human being is immersed in the composition according to claim 2 comprising the bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) as the active ingredient.

5. The method of claim 4, wherein the composition is in the form of a medicated bath.

6. A method for preventing and treating a disease caused by aeromonas hydrophila, comprising:

administering to an animal other than human a composition comprising the bacteriophage Aer-HYP-3 (deposition No: KCTC 13479BP) as the active ingredient according to claim 2.

7. The method of claim 6, wherein the composition is in the form of a feed additive.

Technical Field

The present invention relates to a bacteriophage isolated from the natural world, which infects Aeromonas hydrophila bacteria to thereby kill the Aeromonas hydrophila bacteria, and a method for preventing and treating diseases caused by the Aeromonas hydrophila bacteria using a composition comprising the aforementioned bacteriophage as an active ingredient. More particularly, the present invention relates to a bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) of Myoviridae, which is isolated from the natural world, has the ability to kill Aeromonas hydrophila bacteria and has a genome represented by SEQ ID NO:1, and a method for preventing and treating diseases caused by Aeromonas hydrophila bacteria using a composition comprising the aforementioned bacteriophage as an active ingredient.

Background

Aeromonas belongs to gamma-proteus, is a bacterium that is widely distributed around the world and is ubiquitous in soil and water, and some of these bacteria are conditionally pathogenic bacteria that may cause human diseases. These bacteria are mainly classified into non-motile aeromonas and motile aeromonas. Aeromonas hydrophila is a representative genus of Aeromonas mobilis, and is known to cause acute, chronic and latent infections through oral or transdermal infections in freshwater fish, amphibians, reptiles and humans. Serotypes of aeromonas hydrophila are known to include thermostable somatic O antigen, thermolabile capsular K antigen, and flagella H antigen, and it is known that most pathogenic aeromonas hydrophila bacteria known to date exhibit thermostable somatic O antigen.

Aeromonas hydrophila causes its systemic acute hemorrhagic septicaemia for some of the fish that are affected by pressure due to various environmental factors (such as temperature, intensive farming and organic matter in the water) and because of wounds, parasitic infections and other pathogens, and are ultimately infected. In particular, if an infection with Aeromonas hydrophila bacteria occurs at high temperature, smolt will die in large scale, black bass will develop red spot, and goldfish will develop secondary skin infection and furunculosis. Furthermore, it is known that in the warm water fish farming industry, the high mortality rate caused by infection with Aeromonas hydrophila leads to serious economic losses. Therefore, there is an urgent need to develop methods suitable for the prevention and further treatment of aeromonas hydrophila infection.

Although various antibiotics have been used for the prevention or treatment of diseases caused by aeromonas hydrophila, currently, the resistance of bacteria to such antibiotics is increasing, and thus, there is an urgent need to develop methods other than antibiotics.

Recently, the use of bacteriophages as a countermeasure against infectious bacterial diseases has attracted much attention. In particular, these bacteriophages have received great attention due to their strong antibacterial activity against antibiotic-resistant bacteria. Bacteriophages are very small microorganisms that infect bacteria. Once the phage infects the bacteria, the phage proliferates within the bacterial cell. After propagation, the progeny of the phage disrupt the cell wall of the bacteria and escape from the host bacteria, indicating that the phage has the ability to kill the bacteria. The manner in which bacteriophages infect bacteria is characterized by its very high specificity, and therefore, the range of bacteriophage types that infect a particular bacterium is limited. That is, a certain bacteriophage can only infect a specific bacterium, which means that a certain bacteriophage can provide an antibacterial action against only a specific bacterium. Due to this bacterial specificity of bacteriophages, bacteriophages have an antibacterial effect only on the target bacteria, without affecting the symbiotic bacteria in the environment or in the animal body. Conventional antibiotics, which have been widely used in bacterial therapy, can affect many other kinds of bacteria, leading to problems such as environmental pollution and interference with the normal flora in animals. In contrast, the use of bacteriophage does not interfere with the normal flora in the animal because the target bacteria are selectively killed by the use of bacteriophage. Thus, the phage can be safely utilized, thus greatly reducing the possibility of adverse effects from the use of phage compared to antibiotics.

The phage was originally discovered by the british bacteriologist Twort in 1915, when he noticed that micrococcus colonies were softened and became transparent by something unknown. In 1917, the french bacteriologist d' Herelle found that shigella dysenteriae in the filtrate of the excrement of the shigella dysenteriae was destroyed by something, and further studied this phenomenon. Results he independently identified the phage and named it "phage", meaning "bacterial phagemid". Since then, phages acting on pathogenic bacteria such as Shigella, Salmonella typhi and Vibrio cholerae have been constantly identified.

Because of the unique ability of bacteriophages to kill bacteria, bacteriophages have since been discovered and have received much attention as a possible effective countermeasure against bacterial infections, and many studies have been carried out in connection therewith. However, since the discovery of penicillins by fleming, the study of bacteriophages has only been continued in some eastern european countries and the former soviet union due to the widespread dissemination of antibiotics. Since 2000, the limitations of conventional antibiotics have become apparent due to the increase of antibiotic-resistant bacteria, the possibility of developing bacteriophages as alternatives to conventional antibiotics has become prominent, and thus bacteriophages as antibacterial agents have attracted attention again.

As mentioned above, bacteriophages tend to be highly specific for the target bacteria. Because of the high specificity of bacteriophages for bacteria, bacteriophages often exhibit an antibacterial effect only on some of these strains, even against the same species. In addition, the antibacterial strength of the bacteriophage varies depending on the target strain. Therefore, in order to effectively control a specific bacterium, it is necessary to collect a variety of useful phages. Therefore, in order to develop an effective phage utilization method for controlling Aeromonas hydrophila, it is necessary to obtain a plurality of phages that exhibit antibacterial effects against Aeromonas hydrophila. In addition, it is necessary to screen out phages superior to other phages in terms of antibacterial intensity and spectrum from the obtained phages.

Disclosure of Invention

Technical problem

Accordingly, the present inventors have made an effort to develop a composition suitable for preventing or treating a disease caused by Aeromonas hydrophila using bacteriophage that is isolated from nature and can kill Aeromonas hydrophila, and have further made an effort to set a method for preventing and treating a disease caused by Aeromonas hydrophila using the composition. As a result, the present inventors isolated phages suitable for this purpose from nature and determined genomic sequences that distinguish the isolated phages from other phages. Thereafter, the inventors of the present invention developed a composition comprising bacteriophage as an active ingredient, and determined that the composition can be effectively used for the prevention and treatment of diseases caused by aeromonas hydrophila, thereby forming the present invention.

Accordingly, it is an object of the present invention to provide a bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) of the Myoviridae family, which is isolated from the natural world, has the ability to kill Aeromonas hydrophila, and has a genome represented by SEQ ID NO: 1.

It is another object of the present invention to provide a composition suitable for the prevention and treatment of diseases caused by Aeromonas hydrophila, which comprises, as an active ingredient, an isolated bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP), the isolated bacteriophage Aer-HYP-3 infecting Aeromonas hydrophila, thereby killing the Aeromonas hydrophila.

It is still another object of the present invention to provide a method for preventing and treating diseases caused by Aeromonas hydrophila using a composition suitable for preventing and treating diseases caused by Aeromonas hydrophila, which comprises, as an active ingredient, an isolated bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP), the isolated bacteriophage Aer-HYP-3 infecting Aeromonas hydrophila, thereby killing the Aeromonas hydrophila.

It is still another object of the present invention to provide a medicated bath agent (maceration agent) for preventing and treating diseases caused by Aeromonas hydrophila using the above-described composition.

It is still another object of the present invention to provide a feed additive for preventing and treating diseases caused by Aeromonas hydrophila to act on breeding by using the above-described composition.

Technical solution

The present invention provides a bacteriophage Aer-HYP-3 (accession No: KCTC 13479BP) of Myoviridae, which is isolated from the natural world, has the ability to specifically kill Aeromonas hydrophila, and has a genome represented by SEQ ID NO:1, and a method for preventing and treating diseases caused by Aeromonas hydrophila using a composition comprising the bacteriophage as an active ingredient.

The present inventors isolated the phage Aer-HYP-3 and deposited it at the Korean type culture Collection of the Korean institute for bioscience and Biotechnology (accession No: KCTC 13479BP) on 7/2.2018.

The present invention also provides a medicated bath agent and a feed additive suitable for preventing and treating diseases caused by Aeromonas hydrophila, which contain bacteriophage Aer-HYP-3 as an active ingredient.

Since the bacteriophage Aer-HYP-3 contained in the composition of the present invention is effective in killing Aeromonas hydrophila, it is effective in preventing (preventing infection) and treating (treating infection) diseases caused by Aeromonas hydrophila. Therefore, the composition of the present invention can be used for the prevention and treatment of diseases caused by Aeromonas hydrophila.

The term "prevention" as used herein refers to (i) prevention of infection by Aeromonas hydrophila and (ii) inhibition of the development of a disease caused by infection by Aeromonas hydrophila.

The term "treatment" as used herein refers to (i) inhibition of a disease caused by Aeromonas hydrophila and (ii) alleviation of the pathological condition of a disease caused by Aeromonas hydrophila.

The term "isolation" as used herein refers to an act of isolating a bacteriophage from the natural world by using various experimental techniques and taking a characteristic that can distinguish the bacteriophage of the present invention from other bacteriophages, and further includes an act of propagating the bacteriophage of the present invention using a biotechnology to make the bacteriophage industrially applicable.

The pharmaceutically acceptable carrier included in the composition of the present invention is generally used for preparing a pharmaceutical preparation, and examples thereof include: lactose, glucose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. In addition to the above ingredients, the compositions of the present invention may also include lubricating agents, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents and preserving agents.

The composition of the invention contains bacteriophage Aer-HYP-3 as an active ingredient, 1 × 10¹pfu/ml to 1 × 10³⁰pfu/ml or 1 × 10¹pfu/g to 1 × 10³⁰Concentration of pfu/g, preferably at 1 × 10⁴pfu/ml to 1 × 10¹⁵pfu/ml or 1 × 10⁴pfu/g to 1 × 10¹⁵The concentration of pfu/g comprises the bacteriophage Aer-HYP-3.

The compositions of the present invention may be formulated according to methods that can be readily practiced by those skilled in the art using pharmaceutically acceptable carriers and/or excipients to prepare the compositions in unit dosage form or to add the compositions to multi-dose containers. Here, the formulation may be provided in the form of a solution, a suspension, or an emulsion in an oil or aqueous medium, or in the form of an extract, a powder, a granule, a tablet, or a capsule, and may further contain a dispersing agent or a stabilizer.

The composition of the present invention may be provided in the form of a medicated bath agent or a feed additive according to its use purpose, but is not limited thereto. To enhance its effectiveness, phages having antibacterial activity against other bacterial species may also be included in the compositions of the invention. In addition, other types of bacteriophages having antibacterial activity against Aeromonas hydrophila may also be included in the compositions of the invention. These phages may be appropriately combined to maximize the antibacterial effect of the composition, because the antibacterial activity of these phages against Aeromonas hydrophila may vary depending on the antibacterial intensity or antibacterial spectrum.

Advantageous effects

According to the present invention, the method for preventing and treating diseases caused by Aeromonas hydrophila using a composition comprising bacteriophage Aer-HYP-3 as an active ingredient may have an advantage of having very high specificity to Aeromonas hydrophila, as compared to conventional methods based on existing antibiotics. This means that the composition can be used for the prevention and treatment of diseases caused by Aeromonas hydrophila without affecting other useful commensal bacteria and with less side effects due to its use. Generally, when antibiotics are used, symbiotic bacteria are also damaged, eventually reducing the immunity of animals, and thus, the use of antibiotics causes various side effects. Meanwhile, for each of the phages exhibiting antibacterial activity against the same bacterial species, the antibacterial effect of the phages differs because of the difference in antibacterial intensity or antibacterial spectrum [ i.e., the range in which the phages exhibit antibacterial activity against various strains belonging to the same bacterial species, because the phages usually exhibit antibacterial activity against some strains belonging to the same bacterial species, that is, the sensitivity of different strains to the phages varies even though they belong to the same bacterial species ]. Therefore, the present invention can provide different antibacterial activity against Aeromonas hydrophila from that of other phages acting on Aeromonas hydrophila, which will make a great difference in effectiveness when used in the industrial field.

Drawings

FIG. 1 shows an electron micrograph of bacteriophage Aer-HYP-3.

FIG. 2 shows the results of an experiment on the ability of the bacteriophage Aer-HYP-3 to kill Aeromonas hydrophila. Based on the center line of the plate medium, the left side shows the results obtained using only the buffer not containing bacteriophage Aer-HYP-3, and the right side shows the results obtained using the solution containing bacteriophage Aer-HYP-3, and the clean zone observed on the right side is a plaque formed due to the decomposition of the test target bacteria by the action of bacteriophage Aer-HYP-3.

Detailed Description

The present invention will be better understood by the following examples, which are set forth merely to illustrate the invention and should not be construed as limiting the scope of the invention.

Example 1: isolation of phage capable of killing Aeromonas hydrophila

A sample collected from nature is used to isolate phages capable of killing aeromonas hydrophila. Here, the Aeromonas hydrophila strain used for phage isolation has been previously isolated by the inventors of the present invention and identified as Aeromonas hydrophila.

The process of isolating the phage is described in detail below. The collected sample was added to a TSB (tryptic Soy Broth) medium (casein digest, 17 g/L; Soy digest, 3 g/L; glucose, 2.5 g/L; NaCl, 5 g/L; dipotassium hydrogenphosphate, 2.5g/L) inoculated with Aeromonas hydrophila at a ratio of 1/1,000, followed by shaking culture at 25 ℃ for 3 to 4 hours. After completion of the culture, centrifugation was performed at 8,000rpm for 20 minutes, and the supernatant was recovered. The recovered supernatant was inoculated with Aeromonas hydrophila at a rate of 1/1,000, and then cultured with shaking at 25 ℃ for 3 to 4 hours. When phages were included in the sample, the above procedure was repeated 5 times in total to sufficiently increase the number of phages (titer). After repeating this process 5 times, the culture solution was centrifuged at 8,000rpm for 20 minutes. After centrifugation, the recovered supernatant was filtered using a 0.45 μm filter. The filtrate thus obtained was used in a typical spot assay to examine whether phages capable of killing aeromonas hydrophila were included therein.

The spot measurement was performed as follows. At 1/1,000TSB medium was inoculated with Aeromonas hydrophila at a rate, and then cultured overnight with shaking at 25 ℃. 3ml (OD) to be prepared as described above₆₀₀1.5) was dispersed in TSA (tryptic soy agar: casein digest, 15 g/L; 5g/L of soybean digestive juice; NaCl, 5 g/L; agar, 15g/L) plates. The plate was placed on a clean bench for approximately 30 minutes to dry the dispersed solution. After drying, 10. mu.l of the filtrate prepared as described above was spotted on a plate on which Aeromonas hydrophila was dispersed, and then left to stand for about 30 minutes for drying. After drying, the spotted plate was incubated at 25 ℃ for 1 day for static culture, and then examined for formation of a clear zone at the position where the filtrate was dropped. If the filtrate produced a clean zone, it was judged that phages capable of killing Aeromonas hydrophila were included. By the above examination, a filtrate containing phages having the ability to kill Aeromonas hydrophila can be obtained.

Pure phage were isolated from the filtrate that was confirmed to have phage capable of killing aeromonas hydrophila. A typical plaque assay was used to isolate pure phage. Specifically, the plaque formed during the plaque assay was recovered using a sterile pipette tip, added to the culture solution of Aeromonas hydrophila, and then cultured at 25 ℃ for 4 to 5 hours. Thereafter, centrifugation was performed at 8,000rpm for 20 minutes to obtain a supernatant. To the thus-obtained supernatant was added a culture solution of Aeromonas hydrophila at a volume ratio of 1/50, and then, the culture was carried out at 25 ℃ for 4 to 5 hours. To increase the number of phages, the above procedure was repeated at least 5 times, after which centrifugation was carried out at 8,000rpm for 20 minutes to provide the final supernatant. The thus obtained supernatant was used to perform plaque assay again. Usually, the isolation of pure phage cannot be accomplished by a single iteration of a process, and therefore, the above process is repeated using the plaques formed above. After at least 5 repetitions, a solution containing pure phage was obtained. The process of isolating pure phage is typically repeated until the plaques produced become similar to each other in both size and morphology. In addition, electron microscopy was used to confirm the final isolation of pure phage. The above procedure was repeated until the isolation of pure phage was confirmed by electron microscopy. Electron microscopy was performed in a typical manner. Briefly, a solution containing pure phage was loaded onto a copper mesh, then negatively stained with 2% uranyl acetate and allowed to dry. Then, the morphology thereof was observed using a transmission electron microscope. An electron micrograph of the isolated pure phage is shown in figure 1. Based on its morphological characteristics, it was confirmed that the novel phages isolated as above were phages of the family Myoviridae.

The solution containing the pure phage confirmed as above was subjected to the following purification treatment. To the solution containing pure phage, a culture solution of hygrophilous bacteria was added in a volume ratio of 1/50 based on the total volume of the solution, and then, cultured for 4 to 5 hours. Thereafter, centrifugation was performed at 8,000rpm for 20 minutes to obtain a supernatant. This process was repeated 5 times in total to obtain a solution containing a sufficient number of phages. The supernatant from the last centrifugation was filtered using a 0.45 μm filter and then subjected to a typical polyethylene glycol (PEG) precipitation treatment. Specifically, to 100ml of the filtrate, PEG and NaCl were added in accordance with 10% PEG 8000/0.5M NaCl, and they were left to stand at 4 ℃ for 2 to 3 hours, and then, centrifuged at 8,000rpm for 30 minutes to provide a phage precipitate. The phage pellet thus obtained was suspended in 5ml of a buffer (10mM Tris-HCl, 10mM MgSO)₄0.1% gelatin, pH 8.0). The resulting material may be referred to as a phage suspension or phage solution.

The phage purified as described above was collected, designated as phage Aer-HYP-3, and then deposited at 7.2.2018 in Korean institute of bioscience and biotechnology in Korean type culture Collection (accession No: KCTC 13479 BP).

Example 2: isolation and sequence analysis of the genome of phage Aer-HYP-3

The genome of the phage Aer-HYP-3 was isolated as follows. The genome was isolated from the phage suspension obtained using the same method as described in example 1. First, in order to eliminate DNA and RNA of Aeromonas hydrophila included in the suspension, 200U of DNase I and 200U of RNase A were added to 10ml of phage suspension, which was then left to stand at 37 ℃ for 30 minutes. After leaving it for 30 minutes, 500. mu.l of 0.5M ethylenediaminetetraacetic acid (EDTA) was added thereto in order to inactivate the activities of DNase I and RNase A, and the resulting mixture was left to stand for 10 minutes. Further, the resulting mixture was allowed to stand at 65 ℃ for another 10 minutes, and then 100. mu.l of proteinase K (20mg/ml) was added thereto to destroy the outer wall of the phage, followed by reaction at 70 ℃ for 20 minutes. Thereafter, 500. mu.l of 10% Sodium Dodecyl Sulfate (SDS) was added thereto, and then the reaction was allowed to occur at 65 ℃ for 1 hour. After the reaction was carried out for 1 hour, 10ml of a mixed solution of phenol, chloroform and isoamyl alcohol was added to the obtained reaction solution, and the mixed solution was mixed at a composition ratio of 25: 24: 1 and then thoroughly mixed. Then, the resulting mixture was centrifuged at 13,000rpm for 15 minutes to separate layers. Among the separated layers, the upper layer was selected, to which isopropanol was added at a volume ratio of 1.5, and which was centrifuged at 13,000rpm for 10 minutes to precipitate the genome. The precipitate was recovered, washed by adding 70% ethanol, and centrifuged at 13,000rpm for 10 minutes. The washed precipitate was recovered, dried in vacuo and then dissolved in 100. mu.l of water. This process is repeated to obtain a sufficient amount of genome of the bacteriophage Aer-HYP-3.

The genome thus obtained was subjected to next-generation sequencing analysis using an Illumina Mi-Seq sequencer of Macrogen corporation, and then information on the sequence of the genome of the bacteriophage Aer-HYP-3 was obtained. The final analyzed genome of phage Aer-HYP-3 was 54,451bp in size, and the entire genomic sequence was represented by SEQ ID NO: 1.

The homology (similarity) of the phage Aer-HYP-3 genome sequence obtained as above with previously reported phage genome sequences was investigated using BLAST on the web. Based on the results of the BLAST survey, the genomic sequence of phage Aer-HYP-3 was identified as having relatively high homology (query range/identity: 93%/97%) to the sequence of Aeromonas phage pAh6-C (GenBank accession number KJ 858521.1). However, the number of Open Reading Frames (ORFs) of the phage Aer-HYP-3 genome was 75, while we found that Aeromonas phage pAh6-C had 86 open reading frames, based on which these two phages were also evaluated to be different.

Therefore, the following conclusions can be drawn: the bacteriophage Aer-HYP-3 is a novel bacteriophage distinct from previously reported bacteriophages. Furthermore, since the antibacterial intensity and antibacterial spectrum of a bacteriophage generally vary according to the type of bacteriophage, it is considered that the bacteriophage Aer-HYP-3 may provide an antibacterial activity different from that of any other bacteriophage previously reported.

Example 3: evaluation of the ability of the phage Aer-HYP-3 to kill Aeromonas hydrophila

The ability of the isolated phage Aer-HYP-3 to kill Aeromonas hydrophila was evaluated. To evaluate the killing power, the formation of denuded zones was observed using a spot assay in the same manner as described in example 1. A total of 15 strains isolated by the present inventors and identified as aeromonas hydrophila were used as aeromonas hydrophila strains for evaluation of killing ability. The phage Aer-HYP-3 has the capability of killing 13 strains in 15 aeromonas hydrophila strains in total, and achieves the experimental aim. Representative experimental results are shown in fig. 2. Also, the ability of the phage Aer-HYP-3 to kill Edwardsiella tarda, Vibrio anguillarum, Vibrio piscicola, lactococcus garvieae, Streptococcus paracasei, Streptococcus iniae and Aeromonas salmonicida was evaluated. As a result, phage Aer-HYP-3 does not have the ability to kill these microorganisms.

Therefore, the following conclusions can be drawn: the bacteriophage Aer-HYP-3 has a strong ability to kill Aeromonas hydrophila and exhibits antibacterial effects on many strains of Aeromonas hydrophila, indicating that the bacteriophage Aer-HYP-3 can be used as an active ingredient of a composition for preventing and treating diseases caused by Aeromonas hydrophila.

Example 4: experiment for preventing Aeromonas hydrophila infection Using bacteriophage Aer-HYP-3

To a medium containing 9ml of TSBTo the test tube (2), 100. mu.l of 1 × 10⁸pfu/ml phage Aer-HYP-3 solution. To another tube containing 9ml of TSB medium, only the same amount of TSB medium was additionally added. Then, a culture solution of Aeromonas hydrophila was added to each tube so that the absorbance at 600nm reached approximately 0.5. After addition of Aeromonas hydrophila, the test tube was transferred to an incubator at 25 ℃ and then subjected to shaking culture, during which the growth of Aeromonas hydrophila was observed. As shown in table 1 below, it was observed: in the test tube to which the phage Aer-HYP-3 solution was added, the growth of Aeromonas hydrophila was inhibited, whereas in the test tube to which the phage solution was not added, the growth of Aeromonas hydrophila was not inhibited.

[ Table 1]

Growth inhibition of Aeromonas hydrophila

The above results show that: the phage Aer-HYP-3 of the invention not only inhibits the growth of aeromonas hydrophila, but also has the capability of killing aeromonas hydrophila. Therefore, the following conclusions are drawn: the bacteriophage Aer-HYP-3 may be used as an active ingredient of a composition for preventing diseases caused by Aeromonas hydrophila.

Example 5: animal test for preventing diseases caused by Aeromonas hydrophila using bacteriophage Aer-HYP-3

Two total groups of rainbow trout, twenty each (average body weight: 22.8 g; average body length: 14.9cm), were separately prepared and cultured in a water tank, and the experiment was conducted for 14 days, the environment around the water tank was controlled and the temperature in the laboratory where the water tank was located was kept constant, and the rainbow trout in the experimental groups (given phage) were provided with a feed containing 1 × 10 in a typical feeding manner throughout the test period from the start of the experiment⁸pfu/g phage Aer-HYP-3 feed. For comparison, rainbow trout in the control group (not given phage) was provided with a phase in the same rearing mannerFeed containing the same components but not including bacteriophage Aer-HYP-3, and 2 days from the seventh day after the start of experiment, 1 × 10 was added to the feed⁸cfu/g of Aeromonas hydrophila, twice daily feed, resulting in infection with Aeromonas hydrophila. From the ninth day after the start of the experiment, all test animals were examined daily for furunculosis. Furunculosis is assessed by measuring the size of ulcers on the body surface. The size of the ulcer on the body surface was measured according to the usual Ulcer Size (US) score (normal (no ulcer): 0; small ulcer (ulcer size: less than 0.5 cm): 1; large ulcer (ulcer size: 0.5cm or more: 2)). The results are shown in table 2 below.

[ Table 2]

Measurement of ulcer size on body surface (average)

From the above results, it was confirmed that the phage Aer-HYP-3 of the present invention is very effective in preventing diseases caused by Aeromonas hydrophila.

Example 6: use of bacteriophage Aer-HYP-3 for the treatment of diseases caused by Aeromonas hydrophila

Separately preparing and culturing a total of two groups of rainbow trout in water tanks, each group consisting of forty fish (average body weight: 23.2 g; average body length: 15.4cm), carrying out 14-day experiments, controlling the environment around the water tanks and maintaining the temperature in the laboratory where the water tanks are located constant, and continuing for 3 days from the fifth day after the start of the experiments, twice a day was provided with 1 × 10 stain every typical feeding manner⁸cfu/g Aeromonas hydrophila feed. From the last day of the process of providing the feed contaminated with Aeromonas hydrophila, clinical symptoms of furunculosis in the rainbow trout of the subjects were observed in both water tanks simultaneously. From the second day after the rearing with the feed contaminated with Aeromonas hydrophila (the first day after the start of the experiment)Eight days later), rainbow trout in the experimental group (given phage) were provided with a feed containing phage Aer-HYP-3(1 × 10) in a typical feeding manner⁸pfu/g) feed. For comparison, rainbow trout in the control group (not given phage) were provided with feed having the same composition but not including phage Aer-HYP-3 in the same feeding manner. All test animals were examined daily for furunculosis from the second day after 3 days of forced infection with Aeromonas hydrophila (from the eighth day after the start of the experiment). As shown in example 5, furunculosis caused by aeromonas hydrophila was evaluated by measuring the size of ulcer on the body surface. The results are shown in table 3 below.

[ Table 3]

Measurement of ulcer size on body surface (average)

From the above results, it was confirmed that the bacteriophage Aer-HYP-3 of the present invention is very effective in treating diseases caused by Aeromonas hydrophila.

Example 7: preparation of feed additive and feed

The phage Aer-HYP-3 solution was used to prepare a feed supplement containing an amount of 1 × 10 per gram of feed supplement⁸pfu phage Aer-HYP-3. The feed additive was prepared in the following manner: maltodextrin (50%, w/v) was added to the phage solution, which was then lyophilized and finally pulverized into a fine powder. In the above preparation process, the drying treatment may be drying under reduced pressure, drying by heating or drying at room temperature. To prepare a control for comparison, a phage solution was not used, but a buffer (10mM Tris-HCl, 10mM MgSO. RTM.) for preparing a phage solution was used₄0.1% gelatin, pH 8.0) to prepare a feed additive containing no phage.

One of the two feed additives thus prepared was mixed with wet pellets mainly for raw fish at a weight ratio of 250, respectively, to obtain two final feeds.

Example 8: preparing medicated bath agent

The preparation method of medicated bath agent is as follows. The phage Aer-HYP-3 solution was used to prepare a medicated bath containing an amount of 1 × 10 per ml of medicated bath⁸pfu phage Aer-HYP-3. In the method of preparing the medicated bath, the phage Aer-HYP-3 solution was added such that the buffer used to prepare the phage solution contained an amount of 1 × 10 per ml of buffer⁸pfu phage Aer-HYP-3, and mixed well. To prepare a control for comparison, the buffer used for preparing the phage solution was used as it is as a medicated bath agent containing no phage.

The thus prepared two kinds of medicated baths were diluted with water at a volume ratio of 1,000 to obtain final medicated baths.

Example 9: confirming the feeding effect of rainbow trout

The feed and medicated bath prepared in example 7 and example 8 were used to evaluate the improvement of rainbow trout farming as a result of farming. In particular, this assessment focuses on mortality. A total of 200 rainbow trout were divided into two groups, each group consisting of 100 rainbow trout (group A: raised with feed and group B: treated with medicated bath) and the experiment was performed for four weeks. Each group was subdivided into subgroups each including 50 rainbow trouts, and the subgroups were classified into a subgroup to which phage ae-HYP-3 was applied (subgroup-r) and a subgroup to which phage was not applied (subgroup-r). In this experiment, the rainbow trout of interest was rainbow trout of 5 weeks of age (average body weight: 23.2 g; average body length: 15.4cm), and the rainbow trout in the experimental subgroup were bred in separate water tanks placed at spaced intervals from each other. The subgroups are sorted and named as shown in table 4.

[ Table 4]

Subgroup classification and expression in rainbow trout feeding experiments

In the case of providing the feed, the feed prepared in example 7 was provided in a typical feeding manner as classified in table 4. The treatment with the medicated bath agent was performed according to the following typical treatment pattern: the fish were immersed in a diluted solution of the medicated bath as classified in table 4 using the medicated bath prepared as described in example 8. The results are shown in table 5 below.

[ Table 5]

Mortality in rainbow trout rearing experiments

Group of	Dead rainbow trout/experimental rainbow trout (number)	Mortality (%)
			A-①	4/50	8.0
A-②	10/50	20.0
			B-①	6/50	12.0
B-②	17/50	34.0

The above results show that: the provision of the feed prepared according to the present invention and the treatment using the medicated bath prepared according to the present invention is effective in reducing mortality of rainbow trout breeding. Therefore, the following conclusions are drawn: the composition of the present invention is effective when used for rearing rainbow trout.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that the detailed description is of preferred embodiments only, and that the scope of the present invention is not limited thereto. Accordingly, the scope of the invention should be determined by the appended claims and their equivalents.

The name of the depository institution: KCTC

The preservation number is as follows: KCTC 13479BP

The registration date: 20180207

International recognition of microbial preservation for patent procedures under the Budapest treaty

Issued according to rule 7.1

Receipt of original deposit

International watch

Name of depositor: lntron biotechnology limited address: 137Sagimakgol-ro, Joongwon-gu, Seongnam-si, Gyeonggi-do, Republic of Korea (zip code 13202)

Sequence listing

<110>iNtRON Biotechnology, Co. Ltd.

<120> novel aeromonas hydrophila phage Aer-HYP-3 and its use in inhibiting growth of aeromonas hydrophila bacteria

<130>KP090559

<160>1

<170>SIPOSequenceListing 1.0

<210>1

<211>54451

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<213>Bacteriophage Aer-HYP-3

<220>

<221>misc_feature

<222>(13443)..(13443)

<223>n is a, c, g, or t

<400>1

catcatcctg aaccagacca ctccggaagg tcaggaaacc gtacgtatcc gtgagaccct 60

gccgctgcag ttcctgccgg ttcagctcca cggtttggtg tacgaagttc cgggtcgtgg 120

ccgcttcggt ggtctggaag tgacctaccc gcgcgccatc gacatctggt acggcctgta 180

ataccccgta actgagggct ccggccctct aatttggaga agataatgca agttcaaaac 240

actactgatc actacatctc tttcgttgct cgtaccaatc ctggtgccaa gaccaaaggc 300

tacgacccac gtggtaatga aatcatgcgt gacgagctgc cgcagctgcg tactgtgacc 360

atccctggtg gtgccactgt cgaagttgac gatgcactct ggaacgctgc taccagcggc 420

aagcctgcca cccgtcaagg tatcgagatg gaaaaggtgc cggtacaact gggttccgac 480

gatccgtcca aggaagccaa gtaccacgtc ctgaaccccg tgggtgacgg caaggtggtg 540

cacttcaatc cggtcatgga gctggtgcga gccggtgatc tgaagatcgt agagcacgcc 600

aagtctgaac tgactctgga agatctgcgg aaggcagttg aactcgctca aggctacgcc 660

ctgcccaagg aagtagacga agagaaactg cagcacatgt acaacgtgct ctgtaagtaa 720

ggacaatcaa tgctcaccta cgattcgtgg ctggacaagt tcccgcagtt cgtagctgtc 780

tctgagcctc aatgggcaga gctccagctc gaagcaaccg tagaaatggg cgccgatgtc 840

tttcgttgga atggccaaga tgcgtacgat attgcccaag gctatctgat ggcgcatctt 900

gctgctcaag cagaatcgaa cgaggcgggg gaccacaccc cgctccaacc gttccgcgag 960

aaagaagttg atgatgtgcg agtcgagtat gcagtgtcga gagatatgca gaataacctc 1020

gacccgtatc tggctaccag ctatggtcag cagtacatca agtggcgacg catggcattt 1080

gctggcccac gtgtgatttg aggttactat gatctcatta gcaccagctt ttggatcgca 1140

caccactact caggtgatgt tgcggttgga aattccaggc cagtacgatg gcgacaacat 1200

ctggattact ggtggttatg gtttagcctt tccaataagg gctacgccaa tcccactagg 1260

tgatccagat tacggcgact acggtaaggc tctgaagtcc gacccgtctg gcgaacggca 1320

accagctcaa atcaagattg catctcgatg gaagctgccg aacaattccc tgatggaata 1380

cggaactgag ttgtacaaga taatccggga aggtgactac cactccgcag ggttctggca 1440

ggccattggt gccacagata ccacactgca tccggtagtg ccgctggatt atccaagtga 1500

tatgatgctg atgtacggca acaagctcgt accagtgtcc cgaatcataa ggccaaggta 1560

tggaaaaagc gggtgatacc cttcgagcta tgaagctcgt ggacctcgct ctgggcaagc 1620

caaagttctc ctatgagatg aagcgcaacc agcctaagcc gaaggtcgac agctttgcag 1680

ctgtgctcct gctcgacgag cgtaatccag gccgtgacag gaacgaagtc attgagactc 1740

caactggatt tatcaaccgg acgtctggag ttcggttggt agtgtttcaa atcttgttca 1800

ccgaaggcat cccagatgtc tctcgattcg tctccagctt tatgcggcct gacgtgcagg 1860

acttcatggt agagaacgat ctcgccgtgt tgaagcacga gaaggtaacg aacaagactc 1920

tgatccttga aaccaactgg gaagtgcgcg agagcgtatt aatcgaatgc ctggtacgcc 1980

ggtcgttcga tagcgagatt ggtatcatct ccgaggtaga tgcaaatggt atctacaacg 2040

agggtgatat gacagtcccg atgcatatca atatcaagga accttgatga gcctttcaat 2100

cactaatgtt gttgacgtca acatcctgat tgccccgaaa gctaaagccc tgcaaagctt 2160

cggcaagctg gtattcgtca ccgacgaaag cccgaaggtg cccctcgctg gtaccatctt 2220

ctcctactct ggcatcgaag ccgtagcaga tgaatgggaa ctgacctctg aggtctacaa 2280

gggtgctacc actttctacg gccagaaccc gagcggttccttcatggtcg cgctggcagc 2340

tgctgctact ccggctgttc tgaccggcgg ctccgccgat ctggcagaca tccaggccat 2400

caccgctggt ggctttacca ttcacgtgaa tggcatggct cagaacatca ctgatgttga 2460

cttctccggc gcgtccacac tgaccgctgc cgctgccatc ctggaaggcg agctgactgg 2520

cgttgacgtg accgtgtctg gcaccagctt cgtgctgacc acccagacca tcggtaccgg 2580

tgcgtccatc tctgtggcca ctgccgacat atccggcact gccgccgcac tggcgctgac 2640

caccctgacc ggtgctgtcg tgactgctgc cgttgtgccg gagtctcctg tggaggcact 2700

gaccaaggcc gccgacactg atccgtcctt ctacggcatc gtgctgaaca agaagtggcg 2760

cgacaccgcc caggccattg acaccgccga gtatgctcag gcgtcccgcc gtgtgttctt 2820

caacacctcc aacgaccctg ccactctgga caagaccagc gaagacagca tcgcgtacaa 2880

actgaaaggc atgtccattc agcgcgtgct gactcactac agctccaagc ccaacgagta 2940

cccgtcctgt gcggttgctg gtcgtgcttt cctggtgaac ttcgaaggca ccaacaccac 3000

cattaccctg aacctcaagg tgatgaaggg cgttaccttc gagaagctgc gtcagaccga 3060

gcatgaagca atgcaggcga acaactgcaa cgccatcgtt gacctggccg gtgcattcgt 3120

atactccgac tcccgcatgg gcgacggtac ctggttcgat gctgttcacg gtgtggactg 3180

gctgcagaac cgtatcgaga ccggtatctt caatcggatg tacaccactc cgaccaagat 3240

cccgtatacc gataccggtg tcaccatcat catcgccgag atcgagcagg ccctgcgtca 3300

agcagtgacc aacggtctgg ttgctccggg caataccacc tctggtgagt acctcccgct 3360

gggttacaag atcgtgtaca ttccgactgc acaagtgtct caagccgaca agtccaaccg 3420

tgtctacaaa ggtatcacct ttgaagccgt tggtgccggt gcgatgcaca aggtagtagt 3480

tggtggctcc ttcaacgaat aaggataacg atgaaacagt atagctttta caacgtcgat 3540

cttctggtcg acggtatcct catggaaggc ttctccgact ctgccgcaat catcaccgcg 3600

tctcgtgacg caccgcaaca cggtaaggtg atggatgcgc gtgggaagat ggttgcgatt 3660

acctcagctg acaagtctgg taccgtaacc ttcgacctgc tgcaagtctc tgattccaac 3720

cagtggctgc aggttcgtgc catgcagacc cacgacgccg gtacttccgg tggcactgac 3780

gtcttcctgc cggtgcagct gatgatgaac gataagatgg gtcgcgctgt cgtgaccggt 3840

gttaacggca tcatcaccat gcagcctggt atcgttcgtg gtaccggtgt agcgaccgac 3900

acctgggtta tgcagttcga gcaactgtgg atcctgcgcg gtcagtccga gaacgtcggc 3960

gtctaaccaa catacggagt aaccaatggc ttgtaaagac atgaatcggg ttatcgatga 4020

taatccgatc tatgttcgtc aatggccggc atctgtagcg ttggagaatt taagtgaggc 4080

gttggggctt atgggccccg acttctcttt cttcgtcgac ggatcctacc aattcacgga 4140

catgctgcaa gtgctgcacc gcagcgattc caagcggctc gttgcactgc tgaagaagtt 4200

tgctatggcc gctcgtgtag acggtaaaga gctgcaagaa gcgcagttca accagtacta 4260

cagcggggag atcttcaaga tcttcaaggt attcgccttc gtagcagaag ttaactaccg 4320

agattttttc gagctagggg tacctccagc agagccagac cagcaggaag agcaggacga 4380

gtcgcaagac ccgcagccgc cagcgccgag cttgacccct tagaagaaaa gccgaagacg 4440

ctcgaagagc tgtttcctga ggttgacttc tacttgtatc ggggtgtaat ttgccatccg 4500

cctatgtgct caatgagtga tctgtcggat ggatcattat ctctgtatga cctccatatc 4560

atgcacgaga taatggacct gaaacaagaa ctgaaacaag gagacaaaga tggctaagcg 4620

cccaaccctt tcgattcccc gcaaggcgaa gcacaaaggg cagatgcaga atccgatcaa 4680

tgactttaaa gctgcaaaga gtagtcactc ttatgctgct gcccagtttg acattccaca 4740

aaagccggtg tcaaaaccgc tgcagcgtaa gagtgcctca ttctgggatg atggagaagg 4800

agaggactat gttgagcgca ctatggggcg acccactgag tccatccact ctgaagagta 4860

catcgcacaa gcaactacgc cggaacaaat cctggcgagt cgtatcaacg cggctggccc 4920

tgatgtagca tcggcattca tgtcgcatgt tgccaaggct aagtccctgg ggtcggtaga 4980

ccatacgggc ttcgcgatga gctccaatcc agctgagagg ctgcaataca tggcagggca 5040

aggaggcatc aacctccaga aagtagcatc agctatgcgg cacagggaca agcagtgggg 5100

atcacctgct gcggagggtt cccacgagga agacttctct gtacgtatgg ttgaggctgc 5160

gaagaccaga gcctaccaag gcctgcaaag cccgacacaa cgcgccatgt caatcgtagg 5220

cgcgatgacc accagtgacc agtcctggaa gcagcaagtt gatgctccag atctgataac 5280

ccctgacacc cacgacaagt gggtagaccc tctacagaag gctgtcaacg cggttcagtt 5340

catagctggt gcgtatattg atccgaacaa gctggaaggt ggtgagttcg gacctgccta 5400

cgccgatcta catgggcgcc ttactaaggt gctctcaagg caaatacctg cacggatagc 5460

gaacactggc ttcgagcaag ccacgcgtgg tgctggtagt aagcctgtag gttggaccca 5520

agccttgccg tctgtacagg acgtccgtaa gatgctgccc gtgtctgacc ggatgttcct 5580

caccaacaag ctggataagc tgtgggagga gaagaatggt cgacatgagt caccgttcac 5640

agacgcgatc aagaccatcc gagacgtgta ccgttctggt aacgagggtg gtctgtccaa 5700

gccgaagaac ttcgagtacc gtgagcggca atatctcctt gacgagatgg ctgcatccgg 5760

acttgtagac ccaacccaga ccgtcatttc ccagaaccct ggacgagacg agatgtatga 5820

cttcaaggtc actaccgatg gtggtgagct gatgaactct gcattgcaag cgctgggtgc 5880

aactgccgct gacctcgttg gtgaccctgt catccgggct caagtaaagc gtgctgctgc 5940

tgaaatagct gcaactggtt ctgcaagtac cactgggact gttggagaca acctgacacc 6000

gaaggctgcc ctgctgggta tgcgtgactt cggtggaggg cctatcccgt tccagctgga 6060

gccgtacaag aagttcgagc gtggccgtgg tggtggcgcc accaccagga agaaaaaccc 6120

tgtagcacct gctggcatca agcctggtga tgtaatccaa acagcagcag aggcccaagc 6180

agaagtacgc caacgcactg gtgaccctga cggagccaga gcaaagtatt tcgcagagcg 6240

gccgtggttg aaagccatcg catcggatga tgacaaatca gacccgatgt cggcatggca 6300

gcgcaatcgt cgtggcaaag tgacctcatc cgccgcaggt gccttggcag acccagagtc 6360

tgcaagagga gctttgcatg cagtggtgaa gggggcgctg accaatccca acgacccagt 6420

gtctattggc cgaggctcta tctggactaa gtccggtgat gccctcgaac cagtggcact 6480

ggactggtac cgtaagcacg tcgaccctga tgcctttgaa cctggtctta tctttaaccg 6540

gaacaaggct ggacaggcaa ctactcctga tgctattgct gatggtggtc gccgtaacgt 6600

agaagtgaag tctcgcaaca aattcattga ccccaacaac ccgttggatg ctaatgaagc 6660

gaagactctg cggaagaact acttgcagat gcagcaccag atgtacctca ccggtgcgtc 6720

cagtactgac cttgtcgaga tcctcaggga ccaagagaac cccaatgccc ctctgggcaa 6780

gaacggcctc aacgacaaca acatccgtaa gcgccgtgta gaccgtgatg acgagctcat 6840

ccggcagatg cggccgcaat gggaaacggc aggaaaggca gctgacaaga tcgcaggcct 6900

gaatgaccgt acccagaaga tgttcgccaa agcagtggaa gaaggtaaca ttcagtcctt 6960

cgagaagctg tccaaacgtc acgggattga aggccatgaa gcactcgcag caacgctcgg 7020

gctggcggaa ggcaagggta gtggcggtgg tggcggcggt ggagacaatt ctcgaccgta 7080

ccgtggaaac tacggcgggt tcggcggtcg tgatgctcct actactcttc ggggtggtgt 7140

gcgtggtggc ttagctgcca tgggtcgacc tggtaagatg ctcaacgtag ctcttgctgg 7200

tgctgaaatc ctctgggaca cagcgaacga catcaacgac acaggcctga atctatctgc 7260

gcaagcacgt gcaactggca tgcgtgaaga cttgttccgt aagacacgac tgagtatgac 7320

ccaaggtaaa taccttgatg atgcctcagc aatgtcggac atgtcatcta tcgcgcttgc 7380

caagggaggc ctggaacagg gtttcactga ccgtgctgta aacttggtca ccagtactcg 7440

tggtgccatt actctaggtg acttgcagaa tacaaacctg caagacccgg atgccatcag 7500

aaacctgatg agtcagacag aaagccgtct tcgccgccga ggcgtaagtg aattcggcat 7560

cgcagctact atggaacagt cagggctgaa gtccttactg agtgccactg atgctgctgg 7620

tggtcgtgag tcgttgaaca acgcagtcac gaacatcaat gacaatattc tggaactgaa 7680

gctgacagcc ggtgattacc tggggcagat tgcaggattc ggggaagagt acttcaaggc 7740

catgagctca gcattgcagg acattgctgg ggtgctagcc ccaaatgctg gtggtggtgg 7800

agctcttgtc gaaggacgag ttcctatgtc aacgaccaga gacggtgtca ttggccctgc 7860

catccagtca gccattgatt cagtgactga cttcgccttg cctacaaggg caggtgaact 7920

ggccgacgag atctcattgt tctctggtat gaacggcacg ctacctgagg ctaacagggc 7980

ggcactcgag aagcaattcc gaggcatctc tgcatctcag cgcaacaggg taatggatgc 8040

aatccgcaac gactccttgg ataccaatca actggccaag gacgttcgtg gtggtggcaa 8100

gctagaagtg gagctcaaga ctgatggtat ctctctgact gtgaaagacc agtccggacg 8160

tatcacttcg aaggctttca aaccttatca aggcggagcg gaataacatg ttcggtcgca 8220

ttatgcggtt ggaggcgagg cgggagctca cggcagaccc gttcttcgtc acagacaagc 8280

tgcgcattaa ctgcgaaatc gaaatgtcgt tggcgcagca actggcccaa gccagtatca 8340

ccatctacaa cctgtcagaa gagaactcaa aagctctgac gtcaggtgac catgaggtgg 8400

cctcgaagga caatgctgag tcactgcgag tggcgtctaa agtctttatc cggctctatg 8460

ctggatacca agacgagatg ctggcgaacg gtgaactgcc attggtgata gagggtattg 8520

ttatgaacgc ctacgcgcgt cgtgcagtac cccaccacct cacgcatcta tttgtactgc 8580

cgttggcaag ctcgttcctg agacagccat tcacaccgtt ctctgtgcct cgtggaactc 8640

ccctgaggtt tgtgttggag cggatgacgt gggaggccgg ctttgaagcg gtgacgtttg 8700

atctgccgga ctccatcctg cagcaagaca tgggtggaga agctcttgag ccggaccctg 8760

acttgtacgg cacattgcac aagctaggaa aggcgtattc gtttactttc tctcaaagag 8820

caagtggcat tgggttctac ccgaggatcg acgatagcgc gcagagccac aacgagttca 8880

accatttgca ggctaacgga gaagtctaca acgtcaagcc attcttgctg aaaggcgcac 8940

ctactgttgg tgttgccacg atcagcttcc cgatggtgtt agacgccaaa gtattcccag 9000

gctgggtgat cgatgtccaa gacatctccg gcactcgagg tgacttggcg ttaccctcag 9060

gtggcttgcc tgactactct gcgcttggtg acccgttgtt ctacacggac gacgttgcca 9120

agtacgcagt gctaccacgg tacatgctaa agaaagtcat ccacaagatg gacacctacg 9180

tcgacttgtg ggagacacta gtagttggga ctgttccgac tgctggtgat tatggaaagt 9240

gggagcaaca tggctaacaa actcagagac acaatgatca tctgggagga agagcccgga 9300

tcaggcaagt ggagtgtact cgcatttgac ggtgtagttc gtgaaggtca cagtggggct 9360

gtccaggtaa ctggctatcc tgtggactca ggcttccaag tctcggacca cgccatacgt 9420

cagaaccgta tcatcacact agacaccatc acctcgaata tctcaatgtc agtagctact 9480

agccgcaaga cattcgaaga gtctttcaat gagctgatgg tggccattgg ctcagctcag 9540

acaggcggtg gtgcaccagt atatgatgct ggagaggtgt tcgacccaga gcaggagata 9600

gcttcagctg gccctgcgta ctccgagacc accaaatacg gtcgcgccgg ttacgacaac 9660

gacgctatca atatcaccat cccgtacacc agcattacac ttgggacgat taccaacccg 9720

attgcaacag cgttactctg ccaagtctct atcgacaagg tggacgaagc gctgaacaca 9780

gtcgacaggt tgaacgccct tggcaagctg gtgcatgtgg tgactctgcg tggtgttcgt 9840

aagaactgtg ttatccgcca gtacgatgca atcaacgatg tgaccaactc ctactcagtg 9900

cctatcgcgc tgacactgga gcagatgaac gtggttgacc tgactcgtag tactgtgcag 9960

gtaagtacca actacagtaa cggtgaagtg gtagctcaag agcaggccac tgtccggcga 10020

gtagtccctg aggctgcaac ggtagagaac gccatcaatc cagcggctgt acagctgttg 10080

gctgcaccgg tatctcgtgc ggttcaagct accgctatct ctgacggtga agtaccagac 10140

attgttccgg catcgttcta cgagatggaa caccgtgaag taccattctc cactgggttt 10200

gacactcggt tcatctacga gggcacagag tacacacttg gaagagtgcg gtacaacgtg 10260

actctcggtt gctttgtgac tatcctgcaa tggcggaaag acggaaagta cgagaccatt 10320

gcaagcctgc cgttgcagag cggggtgaat ctcgttaggc agtatgccac caacatgccg 10380

tcattcgtag ctgtgaatac tcaggagcgg aacagcgacc cgtcagcacc agagaacctg 10440

cgactgtata tcatcaaagg gttcgatgaa acattcttag gattgtaatg gacgtcaaga 10500

cgaagacaat tgggactatt gtgtccttcg acccagaaag ccagatggct acggtcaagc 10560

tggcaaccaa cggaaccaac agtactctcg acaccaacta cttcaaccaa gaagggttga 10620

cgctgataga cgtaccggtg gaattcccaa ggtgtggtgc gttcgtaatg acgttccctg 10680

tgaatgcggg agatgactgt atcgtagagt tctatgagtc tggaatagac cactggctct 10740

acgagaacag acgggcgtac aatgtcgtaa atggccggtc agagccagca gcacgcaggc 10800

ggtatagccg taaagacgcg tcctgccgag tggcaatcga caacctcgcc aaccctattg 10860

gtgggttcaa tacctctgga cttgaaatca ggcatcgtaa cggtaaccag aagatggtat 10920

tccatccgga tggagtgatt gaaacgatca gtccgtctga catcaagttc atagccggtg 10980

gagacatcac ctttaaggct gacggaaata tcggtcttga gggctcgaat atcgagtcca 11040

cagccacggg ttccaaccaa gtgaaaggca caagcgtaac actgaagggc caaagcatct 11100

cgatggagca ataatgccag ccgtactcac gcatgtctca aagaccagtg ggcacgatgg 11160

attccctcct acccagtcta agggcggtaa ctcgttcgta ggcattggag gtcatccggt 11220

tatactggta ggccaagaat tcgaaccgca ctctaacggt agtactcaca caccgaagct 11280

gattggcggc tcctcgttca tcaaggtgaa cggagtgcct gtcggactag tcggtgataa 11340

acttgattgt ggcgacacgg tcattgagag cgctactagt ttcttcacag caggaggctg 11400

aatgagtttc aatctcaagc tcggtgcaga ccacgatatc atcgttggga gacaggtctc 11460

gagaaccgat aaactggagt acactgtcca gcttgtcaaa tgcaggctcc ttacctttct 11520

tggtgagtgg gccttgaacc gaggaattgg tgtgccttgg actggagtac tggacaggtc 11580

ttacgacatc tccgcgacga agttcgcaat ccagaacact atccagacca ccgtgggtgt 11640

taagtcactc aattctttat ctctgaaagc ggacccaaca acgcgtctgt tgactgtaga 11700

attcacggcg acgtcgatct acggtgaaat tagcgcagga gtaacggtat gaccggtatt 11760

accgaagctg gaatcacggt caagacttat gacgagtgcg ttcgcagtct cgaacaaaga 11820

ttcaagctgg agtttggcga ctcgttcgac acgactccgg aaagccctga tggccagaac 11880

atcaggataa tggcagcatt catctacgat cagtggctgc tggccgagca agcgtaccac 11940

agttacaacc cgtccgtagt gaccggtact ggactggaca accttgttcg gctgaatggc 12000

ctgactcgta tcgtcgatac tccgactcgt gtcggtgtgt acttcgatgc tactactagt 12060

gctggtgagg ttctccctgctggtactgtg gtagagaccg ccactgacaa aattcaattc 12120

gttaccactc gcagcatcat cctgcctggc gaagtgatcg cgtcgtgcct taccaaaggc 12180

gccatcacga tcttcccagg tgaggtcacg gtgattgcag acgatgtagc ttcagacatc 12240

actgtaacca accctgctgc tggtataact ggcgtagtac gtgagacaga cccgcagctt 12300

cgtgcgagac gggaacgatc actggtacgc gcagggacag cgactgctga ggcaatctac 12360

tcagccgtaa ctgacctgaa cctaagcttt attgcagtgc tggagaatga tactgatgct 12420

gttgttgatg gtattcctcc tcacgcattc aggacggtgg tcgagggtgg tgcgctggaa 12480

gacgttgctg agcgggtcta tcggaacaaa gcgataggca tccaagccta cggggacgtg 12540

aacattgaga tcactgactc cagaggctat cctcacatga tcggcttgtc ccgccccaac 12600

caagtaccca tctgggtaaa ggtgaaggtg aagcgaccga gcaacgttgc catcaacagc 12660

ctgcgtgaca tccgtgacgc catggtggaa cacatcaacg gcctgcaaat cgctacggac 12720

gttgaatggg gcaaactgta cgcgccagcc accaatgctg ctccagcagt gaacgtcaag 12780

ctcatcgagg tgagtaccga cggcgtgacg tacgtacagc aggacatcct gatggggcct 12840

gtcgacaaag ccaaaaccga cgagatcaag gtggtggtcg aggagttgcc atgagcactg 12900

taatcgcagc accgctgacg aagcttatgg acctgctgct gatgcagtac aggaactccc 12960

cgaacttcat caagtacctc tcttgttact ccgctgagct gcaagaggta tacgcgtgtc 13020

tacaacagga aatcactgac cgttactacg atgtagccgt tggtgctcag ttggacgtca 13080

tcggggatat cgttggggca ggcagaacgc tagaaggcgt gtcggtagct ggcaacttcg 13140

gctaccttga ctctgctgaa agtcttggta tgggtcgtga agacaacccg aatatcgggg 13200

gtcctttccg ctctgaggaa gacgacaccg tccaagacat caggctcaat gacgagcttt 13260

tccgcaactg gattgaggct cgaatcatca agaatcgtac cggatgcaat actgaagata 13320

ccatcgcgtt cttcacgctg ctcctgaacg acgacgagct tgacgtagag atcacaactc 13380

cagctccagc aacaacgaaa gtgagtctca agaagaccct cacaatctac gaggcagcgc 13440

aantcagaag tctcgcgcag catatcaagc ccgtaggtgt gactttcatc gttgaagact 13500

tcactggtgt cattgagaca ctgccaattg caagggtggc gtaatgacgt tgagaccggt 13560

gctcacgaaa ctgtgggcag aagacgcact cccagagaat ctcggcgacg ctggcagtta 13620

cattccggca aacccgagtt accccgatca gcagccgaac cagtatgctg taggctggac 13680

tgtgaaccac ccagtggtga agcagccgca ccattggatg aactcttggt tgtactcaac 13740

cgactggcag ctgatggagt ggtacaagtc gaatttcagc tggcaacaag agatcacgta 13800

cgttcaaggt gctgtgatta ttgttgacgg agttcggtat gtggcccaaa gcagcaacac 13860

caacaaggca cctctggcac atccagctat ctggttcccg gcaaagttcc atactatgct 13920

gcaagcccag gcagactacg cggcagtagc cctcaaggtg aataaccacg taggcgacaa 13980

tgacaacccg catggggaga cttacgagtc tttcaacggg atgtctcggg ctgacatcga 14040

cgcggcagta caggcacaag acacactggc acagaaccac atcagccgca tggataaccc 14100

gcatgggctg actccggcgg atgtgaatgt actcgacaaa gacatcggtg gcacattcac 14160

cggccaggtg gctatgacgc gaatagacgt cagtggtggt ggtatccgac ggctttccca 14220

aggattcgag atattcttgg atgctggtgc aaggctgggt attgacacca ctaagatgat 14280

ggcacagaaa gatgggcagg aaatgctctc tgatacgaac tacgcagctt ttcgcttgcg 14340

gaattccgat ctgttcaagc caccattgcc agaccttgac atgcctcttg gagccggatt 14400

gaatgcgtac tctgcgccgc agggctgtgt ggtggactat acgtctactc agacaatcag 14460

ctacaccaac aagtcagggc tcgcagccac agcagcaatc gacgaacctg ggttcagtaa 14520

atatggactc caaattcgtg cagcagcagg gcaaatcctg aacccgtcca acatgcctac 14580

tggtatggta gggacagtgt tcgcaatcgt agatgatgtg cctactgttg gtattggtat 14640

gctggataaa agcaacttgc tggaatactt cagccttgca ctgtctatca gggatctcaa 14700

ggtatgggca tttgctttaa caccatatca aatctcagca cttggagtgt aacagatggc 14760

aacaagacct acacttgcgc gaatttgggc gcttagcggg gcacgaaaag accctgggag 14820

tctgaaatac agtaccggct ggattgggga aataccgacc tttgaggtac tgaacttcca 14880

gatgaaccgg attgacactg cgctgctggc cttcgccgag cgtggtgtac ctgaatgggg 14940

cccggacatc gagtacgtac taggtgctca agcatttgcg tccgacggca agatctacgt 15000

gtctaaggtg gcggcgccta gcagggcact gtcaccgcag aacaacctga ctgagtggga 15060

ggaatcctca gcgcagttca ccatggcaca acacgcggcc atggtagcca agttcgatgc 15120

gcacatcgct cgaatggaca accctcatgg ggttactgct gctcaggctg gtacgtacac 15180

ccaggcccag atcaacgaca agatcgcggt ggtaaaccag aagatcacct cgcacacgtc 15240

agacatgggc aacccacacg gcactactgc gtcacagatt ggcgctgtac cgatcactgg 15300

tggcacatac actgggcccg tggtctttgc gaatgcggag accaagatca acccaggtgc 15360

aggtgaccat gctgtatttg ccgacgcaac tgccgtggga ctcaggtaca acacgatcaa 15420

gatcggtatc gagaagagct ctggtcgggc attcatgcag aacggcgcaa gtaagcaata 15480

cctgctgaac gagccagagt acgttgagct gcggaagacg gtagagaaga actacgcagt 15540

gccgactcca gacctggaac tggacttgct gagtgacatt cacatcaagc aaggaatggg 15600

tttctcggac tttgtgcgtg actccagtcg gacttacacg gacaaatctg gcatactgcg 15660

gacagcagct gccggaatgc caaggcatga acagcaaggt ctgctcatcg aagccaacta 15720

tcgtgagtac ctgtcgattg acgcgccgaa taacttcgcg ggcttcgtag attctacgat 15780

gctgattgaa gggatcatcc gggaattccc gccaggtatc tcaaccatcc tgcagaccga 15840

cagttcaggg cgagacgact tcgtatacgt gtcttccgct ggagtggcaa ctttccgtat 15900

tactgactct ggtggtcaag tgcggaactt tgttatcggc aatatccaag ctggtaaact 15960

gttccgtatt gtggtagtcc tgaccgccac acgatacgcc acgtacctcg atggggtagc 16020

gggtgcagct ggggcaatct cgttcgcacc gacactggag tacagctctg tgcgagtagg 16080

cgggacggca atcaacccca cagcctggtg gtgccgcaag ttctcggtat gggcacgtgc 16140

tctgacgccg gaacaaatca cgactcttta aggagcttaa atgaagggat attccttcag 16200

cgagttggta aagctcgcag aatcgatggt gccgcttaac ggcgggacca tgacaggtaa 16260

cctgacggct ccgaagttct tcatgtctgc tgcacaggat atggcagcta atgctgctgt 16320

acggaaggac tacctggagt cggtgcttga tggctcaggc tacaagaccg taacgatcac 16380

agccacgcat actgtgacac ctggccaata catcccgatt actatccgaa gtctcggagc 16440

tcagggtggt gagatcggga tttcgacggc tactggtggt agcgccagtc cgatgaactg 16500

caacacgttc gttggtaaag tgttcgctgg tggctggtca gaacaaggct cgtgggtgtc 16560

tggaatcttc aacatctact cagtgtctga gcgagccatc gctggcttct ccgctggtgc 16620

tgagatctca ggtgccttcg cggcgtacat cgagtctcag gcgttccctg tgaagattcg 16680

tgtacctcgc gacgcagtgg tcgattacaa cggaggtgct gttactatcg gcacatctac 16740

cttccccgta cgtacagggc cgaactacgt agcaggtacc aagacagttg agctgattga 16800

cttcaacaaa ggatctgggt attacgatag ccagccgtat ctgggccgta gcgtgaacgc 16860

cttgtacggc aacttcgatg atttgaagat cactgctgca cagggtactg ctgctgaatc 16920

tgctgtacgt agagactacc ttctatcctc gttgagcacc aagttcgaca agacaggcgg 16980

gactatcacc ggaagtgtag tatcgcaagg cagcatcgaa gccgcgagct acatgcagat 17040

ccggggtgcc tctaacccga tgttggaaat gcaccagcca ggtaactcag caatcctgat 17100

ctacaagcag accaacacaa cgaacatcag attcggacaa ggtaacgggg cggggggcga 17160

ggcaaaaggg tatgcgtcgc tcggtaactc aggttttgaa ttgctagtgg ggaacctgaa 17220

gacctactat aacaacgatc ggggatggac cgacattggc acagcgatgc tgtacggtaa 17280

cgaggtgcaa gtaagcgaag gatcgctgcg aggcttcctg agtcaacggc agacgttctc 17340

tggctaccac tcgattgagt acggttatgg agcactctcc tctgcgaacc cagaatctgg 17400

ggccaatgtg ttctgggcca ctgacggtgg tacatggaca aggcaatggt tcttgtacaa 17460

taatgggatg ttgcggccgc ctacttcagc tggatggtat atcaacccga acggtgactt 17520

atactcacca agacttggca ccacggttgt ggactgggcc gccgccaacc tgtcggctgt 17580

aaaccatacc cacctggcgc agattgcgaa ctacaacatc gtgcaagggc agcacagcgt 17640

gatcgggtcg tacatctttg cagcgttgaa cccagcccag aacgcgtcgt cctgcaaccc 17700

aggcagcaca gtggcaggcg cccacctgta ccctgcagct tgcgcggaat ggagtcagaa 17760

cagagcttgg aacttccaag gctcctggat gtgtatgggt tttgtagacc cgaacaccga 17820

taaccgttgg gatgaccgtg ccaccctgtg gttccgagtt gcttaaggag caataatgca 17880

agaggaagaa gaaaaggtag aattgcctgt aggtgaagct atccacaaac acgaagggca 17940

gcagatctgc gtgctggaag tgaaagacct ggagtgggct aaccacgaac actcctcggt 18000

caatctcctg gtgcgcttca gccactggcc ggagcacttc ctgccgtaca cgctgaacaa 18060

gtacgactcc actgagcacg gccaagaact ctacgcagca tttattgctg gtaagcttgg 18120

cgagcctgct aagtgcactg caccgacccc ggagcagcaa gctgagatga tgttcccagc 18180

ctacaagaag tcgcagcaag acaagctgga gcgtagaatc gcgccgctgg ctcgggctgt 18240

agctattggt attgccactg aagaagagac cgagcagctc aaagagctcg aaatcaaatc 18300

cgtgaagatg atgcgtgcag tcagtatgca ggacgaagta ttccagtgag gtagtaatgg 18360

aagaacttaa gttcagccag atgcagcttg cagacccact tgcgggtagc gagatggtgc 18420

caattctgca aagcggcgtc aacaagctta ttccagcaga ggtcctgaaa ggtcagaacg 18480

gcaaggacgg tgcagatggc aaggatggca ccaacggcat cgacggcaga gacggtgttg 18540

atggtaccaa cggtactaac ggagttgacg gtgatcaagg ccctccgggg cctcctggaa 18600

cacccgcgcc gatcgtagat gactattacc tggacattta cgattgggct ggcgaccagg 18660

ctatcgctga cggcgcattc ctgaactttg ccagcctgcc actggtgaag aaagcgggca 18720

gcactgccgg aactactatt gccgccaatg cagtaaagtt ccctaccaag aacaaaccgt 18780

catccgtagt gtttgtgctg cgaattaccg gcaccatcgg tggctctacc tcccaagcaa 18840

gggaatggcg tatccaaatg cgccgtcctg acgggacagt tgttggttct gtcgccgact 18900

taaagatcac cggcacaact attgtcaatc gtgatgtcac tatcactagc cacacttggg 18960

acgcgaacga cccgttctcg gtgacaggta tccagctggg cttgcacaac gtgtcaaccc 19020

aggcgatgac tatcacggcg ctcagtatcc gtgtgcatcg caacgtcaat ccggagtgat 19080

catgacaatt actgaatgga tggagaactt tgcagctgcc gtagcatctc gacagtacaa 19140

catctacaac acccagacca tcaaggcact gaaacaagct gccgatagcc tgtctgctga 19200

agaatttgtc gtcaaggcaa agcagctgtt cctcccgtac gagcaaagta acatcacctg 19260

ctcgtccctc accggccact accacccgca ctccgcgaga gcactgtggt tgactctgga 19320

caccatcgaa atccctgtag cagggcgtat ggtctccgaa gacggtgcgc cgctgataag 19380

cgaagatgag aagcaactgg tagcagaagg caacacacct taggataacc tatgactacc 19440

ccagttgctg tcggagaagt tgttgcacgg agtgccacct tagagaaggg cggtctcttt 19500

gtagtacttc tcgtagtcct gtatatgggc tacaacatct ttcaaaccac cagtgagtct 19560

gacaagaacg agcaaaagct tattctcact caactcacta ccatcgccca acagcagatc 19620

actatgcagc agtccctgac taacactcag gcgtccgtcc aggctagcca ggatgcaacc 19680

cgtgatgtac tgcgcaagat ggaacaaatc gaaagcacca tgcttcgcta cgtttattac 19740

aacgacgcga acggcacctt ggtcatacga actccagatg ctgttggtgg ttctgaaatc 19800

aagctgaaaa ccgaaccgaa aggagcacga taatgccgct gctgaaagca atcctctggc 19860

cgatcctcaa aaccgtcggc accaaccttc tcatcaacct gatgtctgag ggcatcaagg 19920

agctcgagaa gcgccccaac tcctctgcta ccaaagaaga cgtcaaagtc gtcgaaggta 19980

tcaagaagct ggtgaagtga tatgagccag tctcgccctc acttcaaaga gtctgagtgg 20040

aactgcaagt gtaccacctg taacaaggcg gtgccgcaca agatggacca ggcagtaatg 20100

gataaggtgg agcagctccg agtcctggct ggtcgtcctc tgagcctgtc ctccgcgtat 20160

cgctgcgcca agcacccttc cgaagcaaag aaggcaactc ctggtcagca taacaagggt 20220

ttggcagttg acgtggccgt cgctgacggt gctcaacgca tgcagatcat caagctcggt 20280

ctcgaactcg gtgccaccgg tattggcgta gcaaactcgt ttgtgcacct tgactggcgg 20340

aaatccacac ctgtcgtttg gacttattcc taagtctttt cgatagaaat tgtaacaaaa 20400

tgacccagaa atgggtcgtt ttctaaggaa atcaatgcaa aagtacatga aactcctggg 20460

aatcttgatg cttttgcata tatttcctca gaaagcgatg gcttcgaaag gccatcatta 20520

tgacaacact cggtgttacg cccaggcggc ctaccacgaa gcccgttcca ttgacagaac 20580

ggccatgagg cttgtccaga gcgttgtata caacagagtg aagatgaagg gataccctag 20640

ggatgcctgt ggcgtggtat ggcagagagc ccaattctcg tggactctgc gccatccgca 20700

ctcaatcggt gccctgccga cagccctacc gcccgctgac ggggcggctc tcaaagaggc 20760

actctccctg gcccgctctc tgcgcgccgg tgactggcgt ccctccataa cggcagacca 20820

tttcttgtcg ccagacgcgc tctctggggc caagacgggc aggccgcgat tcccagcgtg 20880

ggcgcggtgc ccagcggggc agcctgaccg ctgtatgtca taccaggggc taaagctcct 20940

agcgccggac ttcacatatc gtggtatctg gttttatact atcaactaag agataacgca 21000

aatgaaagaa caagtagtct acatcgtgat gtatgaagat aatggccaga agtatgttgg 21060

caatccgtac ctcaatccta tcgatgcaaa tgaagaagca gaagagcgcc gccaagtact 21120

gaagaatgca ggagcatctt gggtagtaca gcgcaagatg cggagtcaag aatgcagtta 21180

ggatcggtcg acctcgtcag agggagatag atagccttat taagattcta ttattgagta 21240

accaatgaga ttcttaataa gttagttaat aagattcttc gtgagtcact aatgcgaaga 21300

atgcgaagaa gaattccagt cgaccttggt ggagtaactt agtggcacag acgtagattc 21360

cttatgagaa aaaatttttg gacccatggt acctttatgg aaattatgac gaaaaaattt 21420

tttggccaag aatagggtgg gggaaggatg ttgttgaatc ttgacttgat gggacccgtt 21480

cccgacttga ttccaacgta cttctatcat actcaataag gaatactact atgtctatca 21540

tcaccatgaa caacactgct gctgccacct tcactgacac catggttgaa gagcttgaga 21600

tggttctcat ctccatgctt gactctggta ctatcactga agagcaagct atgaccctca 21660

atgagtgtcg tgatcttcgt tcactgcgta tcaatgctct gaacatgggt cttggtgaag 21720

agattgtcca tgttgttaag gtaatcactg gtgagtatgt caaccagcct ttggttgatg 21780

gtgctaatgt tgatatcagt gagttcttta agtgtaatac ccctgaggag gctaaagcac 21840

tccaatcttg ggctagtgtg tttgatgttg ttatccctac taccatgtca gttgtcgagg 21900

agaaacctat gaagaccgcc aaagacgtaa gccgtgagtt caaagccatc gtggaagctc 21960

gtgatgccat cgaatccagt cagattcgct tggatgacat tgagctcaag agtgccggct 22020

ctatcgctac tgctctcgtt catgcattgg agaaggatgg gttctatacc cagtacttgc 22080

cgctggatga gaatggtcag cagactgata gtcactatca ctttgaggcc aaggcaccga 22140

acttcgacaa tcgttctgtt gctctgcttg ctgtccagct cgctggtgtt ctgccgtaca 22200

ttgacgaaga atacgacttc gaccatgttg tgcaaacagc tgtgttccgt cgtcgcttcc 22260

gtgacattgc tcgtgctaac actttcatcc ctggcgctta ccacaatatg atggatgctg 22320

ctggtactac tcactggcat gagaagctgt ccaagggttt ggagctagct atcaaggctg 22380

gtctgctgga agagcgttgc gaagatggta tcacctacat caagcacact aagaagtaca 22440

ttggttcttg catctctcgc actagcgtga tgcatcagtc tgaagccatc accactgaaa 22500

ctcgtcgcaa agagcgtgtc aaaggtcgct ctaatgctcg caaggatggc acttctaagg 22560

aagttcgtga ggcactggaa ttcatcgagt cccaagctca atgtgttaac cgttggttgc 22620

tggatgcaat caatggcctg attggttact gcactactaa caatctggct gtgccttctg 22680

tgttcaatga atctcgtcat gtgattcatg gctccaacga gctggctaat gttgctgagt 22740

tgttctctga gtacttccat gaccttcgtg gtcgtatgta ccagtttgct cactgtggtc 22800

ctaacccgca agcatctgat atggctaagg cattgtgcta ccacactgtt gttaaccctg 22860

ttgctaaggg tagtgagcag tatcacatgt tcctgaacga aatgttcggt gaagtgatcg 22920

gtgatgctga tagtgtgtgg gccactgagg cttacattcg tcgtactgct gaagagcctg 22980

tcaaagctct ggtacatgca ttcaatacca acaatggtga gcttccgttc aagaagttct 23040

tctcctacat ggacatgtgc cgtacttggg tgagcttcga agacactggt atgggcgaga 23100

gccgtcttgg ctttggtcct gatgctaagt gctctggtgc tcaaatcttc tcaatccttg 23160

ctggttgtaa gaccatggca gaagcatgtg gcctcatcac tggctatgag cagaagcctg 23220

ctgacccgta caacatgtca gctcaggaag ttaacaacat cactcgtact atgattgacg 23280

ctggtcttat cccgactcgc acaatcactc gtaatgagat taagactccg ttcatggcca 23340

tccaatatgg tggtggtgtt ccgtcactgc gcttcaagaa gttcgagcct actatggaag 23400

ctcttggtat cgccagtgat cgtcgtaatg agttctgcca agatgttgtt atcgagggta 23460

tcaacaatgc aatgggtcca gtcattggct cgttcattga agttcttcgt aagtctgctg 23520

aggcctactg cgaagagcac aatgttgatt acttcgagca tcgtcacatt gatggcttcc 23580

tgtgcaccaa gaagggtgaa gctcaagtcc gtatgacttc tgagccgttc atcatcaact 23640

atggtgctga tggtaagggt gttatctttg gtagcaaaga gaaggctact ggttggttgg 23700

tagagtctcg tacttctggc atcctccagc gtcgtaactt ctgctactac ttcccggtac 23760

acttcatcca aggcctggat gctgtgatgg cacgtaagat tgctcttggt gctaagaaag 23820

ctggtcttcg tggttactcc actatccatg accagttccg tgtatgtctg gaagatgctc 23880

ctcgtcttcg tagtgaggtt gttcctcagg tatacgttga catgttcatc aacaatgacc 23940

ctgtaactca tctggccaac cagatgggca ttgagatgaa gtggggtaac ccgctcaagc 24000

aacgtgtgca ggtgctgact gaagaaatcc tgttcagcaa ggatgcctat tacttcgaat 24060

aatcgaacaa gggaatagtc tgttggtaac tcgaagccat tagcaatagt ggcttctatg 24120

gtactagcag cgctcgctta gagccactct taccattgat atgaagtagt gcccaccgtt 24180

gccacagcaa ccacacctct agatgcttcg ccacgggagc ctctcaaaga gaagaccagg 24240

ctctagcgag ccacaagacg ctagcaatcg gctctacgag ccaaaagctt ctgacgatag 24300

ctctgtcgag ccacaagtca ctagcgacct tccattgata tcctcagcgt atagttggtg 24360

gatgcaaggg ttattgctat agtctattgc caatataata actaaccacc ataaccatgg 24420

ttacggccct ggcgggccaa aagaagatga ggcattggta ggctaaccga tgtatacgca 24480

caggcccaac cacccaacga agcagcaacc gtgtaggttc acttctacgg gttagtaagt 24540

cacggcgtct cctcatcgaa actagggcat cgtacccgtg taatgaagac aatgaagact 24600

tgtatgatgt gctgttccac ctacggaaca atcaccgtgg aacgtgagat actaccgtag 24660

agcacatatt tctcctgatt cttattaagg ccgtgtcggc ccaagggtca cgataaatat 24720

tatcatcatt tcaagaatcc ttgcatagca atactacttg tatagatggt atgactgtgt 24780

agagaagaat aatcgtatga gacgtagtat gagacgtccg tatgatgtgc gagtcgaaga 24840

ctcttagtaa gtacacccac agtatgagac gtaagtgggg tgtgtgtggg gacgtggaag 24900

gcaagcggaa ctcgagcgcc acaagacacc agcccctacg tcattcacag acacaggtat 24960

gatacatgta tgacctacat aggtatggtt attacgttac ttactaagag cctacggctc 25020

ccatagggct cttcgagccg aaagtctgtg ataggcatat ggctctacga gccttttgta 25080

attgatgatg ataatattat tatcattacg tatgtctatt attaattatc atgatgctaa 25140

tgtcgcttgc gatttgtagc ctaacctagc ccacaggagg aaccatggca aaggtgctaa 25200

actacgctcg agtacgtcgc atccctgagg gtgcggtata cattggtcgt ggaatgaagg 25260

atatcccaga gaatcctttg ttcaagaacc cgttcaagat gcacaacgag tcacaacgcg 25320

acttggtgtg tgaacagtac aagcaacacc tatggaagca aatacaagct ggcgttgtaa 25380

ccaaagagca cctcatggct cttgatggca aagacttagt gtgctggtgc gctccaaaag 25440

catgccatgg tgacactatc ctgaaggcca tcgagtgggc caaggtgaac atgtgagaac 25500

atcatacagt caccacgtgt acaagaacgg ctcatcgatg gtgaagtcga tatgtcacgt 25560

caccatgcaa ctgacaacgt cgttcaacag gcaagagcgc aggtggaact acaagtgggt 25620

aagaatgcga actgctaaag ttgttcgttg tgcaaagaca ggcgtagtat tatggagtta 25680

ctccgttaac cacgctatgg gaattagtat tgacgatatg ggagaatccg atgccagcta 25740

aaataagtcg tgcagacata ctgagagaca tccgtggtat ctctccagca aagctgggca 25800

agttgcgggt aatcgtggag gagcaataca cgcctcggga ctggttggcg caacgttacc 25860

aagtatccct ggtagtatac cgagggcaaa acaagaatat aattgcccgt gaaagtgatc 25920

tggtaagtcc cgcgcatgtc atccaaggct ggatgttggc ttgtggactc accgccgatg 25980

atgttgaact gaatatcccc aagtaagcaa taggaggctt tatgcccaag ttcatgagta 26040

tcaaagacat gcaacgcctt gtgaaaccag cgaagtacta ctactcgttg acaccacaag 26100

gtcgtgcgaa aatccacgct cagcgtatcg agaacgctat gatggtgcga ggtgactaac 26160

atgtgttggt tatcatttgg ttgtttggtg ttattggtag ttattattta cctttgtggt 26220

gatgataatc tctccgataa caattatcct gagagccctt gggataaata aggctcttcg 26280

agccgcaaga gtatgctcca caccgtcatt gcagtatcgg gtaccggcta tcaagccttg 26340

actgatgcgt gagtctcttc gggtggagca cccattcctt tatggagtta acaatgaaat 26400

atttacttgg tctggtaata gtactctttg cgattgcgtt agtctcggtt atattgatga 26460

ccgtggcaat tgccttcggg taccaaggca tatcgggcga acaatgggct gcggctgcaa 26520

tcctcacgtt catcctttgg atcaccggaa acatcgtagc gagtgtatgc taatggctaa 26580

atttattgca cctgagaaag gcggcttcct gtcctactac gggctgtcga agtaccaact 26640

caactccatc ctgcgtacca atggctacgt gcaggacatg cgattctact ccgacgagga 26700

taactcgttc gtgctggagc gtgcgggtaa tcccaagaca ttcgcaagac tgttctggac 26760

cagcgtaagc ggctggacac tgcacgtcaa gctggaagac gacccgctgt ggatcgtccg 26820

tagcgaatag ctaactaggt gccattcatt gagtggcatc tgtggtagtt atttctgact 26880

atcattttat tggagaactt tatgtacacc aacgaaatgc ttgaacaagt tttaacggca 26940

atgctagact ctggttcaat ccatgaggac acctgccacg ctctgctcgc tactaacacc 27000

aagatgattc cccgtcttgg caagtttgtt gagcaaggca ctctgtcaat caagactgcc 27060

aattccatca tggctggtct gcgtcagctg agggccaaca gatgagtagc cacctagttg 27120

atgtggacta ctcgcaaatc gaaagccgtc tactggcctc caatcctgag tggttcaaag 27180

accagtattt cgcgcattac agagatgatg cgaagtactc ggttaaactg cgttacgaca 27240

tgactccagc agcagatact catatcgttg actggggcca cctcatgcgc atggttcaac 27300

cgattacagc aactggcgtc aacgagcttg caagcttgaa ggccaaccag attgctcaca 27360

ccaatcagta cacgattgtg agggtccaat gacaatgata ttccgagtcg caatagtcgg 27420

cggccgtgac ttcactgaca tgactaaact gactgcatgc tgtgaccaca tgctgagcaa 27480

taaggtaaac ttaggctacc gaatagtcat aatcaatggt actgccaaag gtgctgatga 27540

actcggaggc cgatacgccg acttgcggaa ctacactcaa gagaagttta tacctgagtg 27600

ggaaaagctg ggcaagcgtg ctggcatggt aaggaacact gacatggcca accgtgcgga 27660

tgctatcata gcattctggg atggcgtgtc aaagggcact cgaaacatga tccagacctg 27720

ccaaaggctg ggtaaaccga ttcaagttta caaatactaa ggagccaaga atggctattg 27780

aagttgtaga atacgtacgt cgtgtcatcc tgcgtaacaa ggtgcgtcaa ttcaccctga 27840

accagctggc agacaatttt aagtgtggtc agttgaaggc caaggtcatc tactacgcgg 27900

tagtgactgg ctgcaactct ggcaccatta ccaagctgac catcagcaac gatgactacg 27960

ctcgtctgtg ggcgagggac cgcgacatcc tcaacgtgat caaggagctg taatcatgcg 28020

cgacctgaca tcctgggaag taatcacagt tgtgatctcc gcattcgctt tgctggtgtg 28080

gctctggttc gacctcaaga agttcctgct caagccacgg gaaattccgc agtacatgga 28140

agccaccctc ggcttctctg acgcgttcgt tccgtccaag cacaccgtgt ttgccatcct 28200

tgactggaga catggtgtgg tctcgatgtc attgaagttc gccggcctcg gcagcgccca 28260

cgacatggag atggctagct gtcatgaaca caatttcggt tacactcatc tcgcagaaca 28320

attcgtgagc aacgcgatta tgtgcagcat gagtgtagat ggtatcaatg aacagcaagc 28380

cctcgctgag gtgcgtagct ggaacatcgt agtaagctgt actggtcggg cgcacaatgt 28440

attcccgagc atcaacggcc agcatcctgc ggcactcatg caagatgcac tgcggcatgt 28500

tcgcaatggt tggagtcaat ccaatgcgag tcgtaatttt aatactgctg gctagcctgt 28560

cagcatgcca tcctatattg gccaacatgg acaaaggtga ttgcaccaag agtgcacttg 28620

ctgaactcca agaaggccac aagcggatcg ctgccggtaa aggtggcgat cttggtaaga 28680

aaatcgctaa agacgcgttc gaccgttaca tcgaagagaa gtggttgaac tgcaaaattc 28740

aggagaatca gaatgatcag tcaagagaaa ctcgccaagc tcactaacac tgctgagctg 28800

gccaagaaag tttgtgcaat cgtcaacaca ctggccgtgg gcgaagcctg ggtggccggt 28860

ggtatcgtgc gtgacctgtg ggccgcgcag accaaaggtg ttgacctcga aggcagcggc 28920

gacatcgacg tcctgctgat caactgcccc aacgacgacc tgaaccagct gggtgctttc 28980

atctctgctg ctctggacat caagggcatc aaggcgtact gctcccgtgt ggactatagc 29040

tacgaagcca cagaagatta tccggatgca gatgaccgtc tgtacggcgt gatgcagttc 29100

agcatctaca gcagcggggc agcggatgca agcaagatcg aaatggacat cctggtgtac 29160

gaagaccgtt tcgagacgtt ggaagacgtc atggacacct tcaactgcaa catcaacaag 29220

ttctggttcg atgaagaagg caacggcaac accaccttct ctccggagca gcccttccag 29280

taccgcgaag gtgaccgtga gaagaacgaa gaacgctacg agaagtgcgt gatgcgttac 29340

aacatggtgg ccaaggcagc tcacccgtac cagcgtatcg atgagcacga gggcatgacc 29400

gccgagcaag tcgacagcca attgccctgc tgaggtgtaa catgattgta cctcaaaaag 29460

ggtatgtgta cagcatctac cacagtcgtc gtggctggga gaagctgacc gtctgtgaga 29520

cgtctggaaa cactgtgaag atggtgttcg aggaagggtc gatgaaaggg actgtgcggc 29580

ggatgagcat cggctgctgg aatgaccttg tcaagggcaa tcgcctccaa gagggttgga 29640

ccagcaccac gccgactgtt gaccagaagc tggcggaaat cctgccggta gaagtccaca 29700

tcgaacattc ccatgaaaga ctggaagcaa tcgctggcat accgggcgca atccgccatg 29760

ctggtgagac ccgtgatgaa ttcctgagcc gccttgagcg aacacttaaa cctaaggaga 29820

aacccttgaa acttcgtatc gtagatatct actgttttga caccaccacc aaggacctcg 29880

acgctgccaa tcgcgtgatc gacaaaatcg aaggccacgt gaccgacaag agcgacaacg 29940

acactgttgt tgacctgctg gctgccggaa agttcgtagg cgccctgagc gcccacaacg 30000

agagagcggt tgacgaagag tcgaagcacc tcaagttctc cgacatggaa atccgtgtcg 30060

tagagcgtgc tgcggtgtaa ggctcttcga gccaaaagtt agtgctaaag tcgaagccat 30120

tcaagtagtg gcttctatcg ttatcactac aacaaaggag tgagaatgac cactagagat 30180

tttgatttca ccagtggaat gccgcccggt actaacttgc gtgtgaagtg gccagtcacc 30240

gctgcacaag taagaaccac cggtacgctc tgccattttc gttctccgca agggcacgaa 30300

ttcatgcgca tggttatcgc tagcgccggc caatacctca cgtgggtaga cggtctggag 30360

cctgaatggg ccatgccgta tgcaggtgac gacatgtggg cgactttcaa tgacgggttc 30420

ccccgtcgag tgtgacacag cctaaggaga atccaatggc gaaagattac ctttgcggtg 30480

tagcagcagg tgttgattta gccgtcaacc gtgctgtgcg ccgtgcagta acccacgaga 30540

agggcatcaa gttcatcttc cgtgactcga tgaccggcca agatcgtgag aaactcattc 30600

ccgtaactgt ggatgaatac tgcgactgga ttaatggtag cctgattcag gacgccatgc 30660

cgcatatcca ccctgatgac cgtgagttgt ttctcactgg gatggtgtgg taatgaatac 30720

ctggttcgta ctgttcatca cactgaccta cggcggcagc catgagttcc gtactccgta 30780

tcaagactac cgctcctgcg aagaggcggg ccaacaagtc tacaacatgc ggtacgtcaa 30840

ggacttcatc tgtttgccag taaagcgcta acgcgccaaa agagagtgac taagtaactc 30900

atcaactacc tgctaaggag gatgatatgt agtcactcat gttcaatcaa agggccttcg 30960

gcccacaaga aattcctgga gcaagtatgc aagctatgtc tacccaggcg agcagcatgc 31020

ctgctccgcc gattttcgcc gtcatcccgc gtgtacctta tgacgaggta tgcccgtgct 31080

gtggcactgt tgtgcatcat ggttactgtg gtgcatgtga taaaaccatt gaaggataag 31140

gacatgcatg gtgtcaatgt ctctcgcgtt ttgggctccg gtgcaattga ttttgctact 31200

gaccctctgc gtattggtgt ttgtaaaggc tccccaatgg gccgcctcat tcaccgcagt 31260

aattcccgca aaaccgtggt gcgacgtgtt cagtcgtctc ccgctcatgc gtacaactac 31320

accatcatgg gctaacccaa caggcctctg cgagcaaatc cggattcgga aacagacgtg 31380

gaaaagttga aagttagccg atggtgtagg cactacaacg ggctccggaa cactaccgga 31440

gtcctacaat tagtactctt gcgctggacg cttggagata acgctgaaag aaccaaatgc 31500

gatttggtag aataggtata atgccaagta tgataaagcg tgagagtcct aattatactg 31560

agtgggttaa aggatggcac ttgtagacag ttcgtctaga ggcccaggac aggcgctgcg 31620

cgatgcgtca aacacaggtt caactcctgt actcctatga gtgtcattgc tttaaccctc 31680

ccatcaacgc aacaaagaga atcactctat gcgtaaagaa gacaaagtag tatccctgtc 31740

caccgctact cgtgccgagc gtaagcatgc caagcacgta gcgcgtaccg aagcccgtcg 31800

tgttgccaag tgcaagccga actggcagaa ggagtggaag aaccacatgc tgcaacagcg 31860

cggcatgaac aagatcatgt ccctgatcgg tcgcggcatc gccgtacaga aggtcaagaa 31920

cgctaaggca ggtgtgtaat gagcaacgtc atcaactacg aactcgttcg cgccggtaag 31980

gtgatcttct cctccacccg taagctggac accatctatg ctcaccagaa gcaccgccgc 32040

agcgctggtg tgagctaccg tgaaaccaag gccaagtaag gtatacatat gccctattta 32100

ccgattcatc ctcgtcgtgc gctccgtgtg ggcgacgagg taattcacgt tggtgaaggc 32160

gtgacctacc ggggtcgatt gggccgcgtg gtatcagtat caggtagcaa ggccagtggc 32220

caagtgaaga tactctggga ccacaagatg caggcgctgg actacagcat aggctggtgt 32280

atcaggatgc ttggcgtgca cgtgaactgc ctgcgagtca attaccaaga cttccatggc 32340

aagacttacg tggacaagta tctgcactgg gcgttccaga agggcgacgt caccaaaggc 32400

ttcttcctga ctccgcatca cacgatgaag gccaaggaag acgcgaagct gcgtgccgaa 32460

gaacagtgcc gcgtccagtg ctgcaaagtt gacgtcacga gcgactacca gcgtggtctg 32520

atggcccaac tgaagcgcga ttggtccatc ctccagcatc ctggcctgta caagctcatc 32580

ccgactaccg atggtctcgg gtatcgagta gaggccatcc ctgccaagcc ggcacaacct 32640

gtacaggaga agcctgtgtc caacgtagta taccctgtga ttgccgagag ccgcgagaac 32700

ggtcaaatca ttcacattgc atccgagaaa gagctccaga agctcggtgt cggtagctat 32760

acgctgtact cgaagtcctg caacatcgca gttgctccgg tacagcagaa ggaaatcact 32820

ttcctgtaag cataactcga gagtcctgac cgcctttgaa cagggctctc tatggtatgt 32880

ttaccagata gttttcgtca gtaacacgcg tctgttttta tcatcaaata aatgcaaacg 32940

atgatctgat gttggctgcc gcctgataag cgaaacgcca cgaggtattc tagttcctcg 33000

ccaagcaatc tagcggagtt cctcactctg attaatacag agggcataac tcgagtgtcc 33060

tgcacgaata cagcagggca ttctctggta tgtaaccaat tcggcacttc gtgccacaag 33120

gagttgatca tgtcttacga tcgtcgtaac gaccccaacg atggaatagt gtctgccgag 33180

atgtctctgt gggacttcct cggaggacta ctcctttcat gcgtactagt attccttatt 33240

gtagggtact gcgcaaaata aactgtgtat gtcaacttgg agaaaccact atgacaacca 33300

acctgcaaac cccgaccctc cagcaccccg acctggctcc gcaacaaccg cagaccagtc 33360

agtaccctgc tgagatcgcc aatttcactc tgctcgctgc ccgtaaccgt accagtgaac 33420

gctcaccgca ctacttcgga tccttcaaga tccagggtgt ctggtaccag gtctccacct 33480

ggattgtgtt ctcgcgtagc ggtggcgagc agcaactgtc caacagcatc cgtccgtgca 33540

ctccggaaga agctgccaag catgaagctc gtgaccagca attccaagcc ggtcagcaag 33600

cacgtgctca gcagaccctg ggccaggtgc ctaatccgat gcatgctccg gccaacaccg 33660

gcacaccggc tgtggacccc aaccaccctt tctgacaaag gatcctgccg tgaacaagtc 33720

actgcgtaaa ctggcgagcc tcttcggggg ctccaatatc ccccaggctg gccgtaacgg 33780

tcggcttgac atggaccgcc atctgatcaa agagaccatc ttccgccaat gcgatgccca 33840

catcgacttg aagcggatta ctcgtggtac ctttatacac gtggtgaagg caggtttcaa 33900

atcctctgtc gtccgcctgt tcgaccatga gatcgtggtc agcaacaagt acatcaagca 33960

gtcttaagga gaagccgtgt ccaactttac ggatgaccaa gagatagtcc tcgagattat 34020

caagcagtgg ttacgcaatc aggacgcggc gtttcttgcc cgtgaagaac tcgttgtctt 34080

ctggggaccg attgatggca acttgcgtaa gcaagggtgg aataagctga agctcaacga 34140

ggctgtcagc gtaatccgaa ccacgaaagt ccctgtcggt gtgatgaaac actgcacagc 34200

tgacttgatg cgagcggctg ctcaagaaga aggtcgtgcg tatatcgctg gagtgactgt 34260

tgtagggggc gctgtgagcc cggaatactt caactttcac tctaacaagc ggtctgtatg 34320

cgagaccatc gagcaccgtg ttgccattca tctgatctgt gagcttgaag ccctggacca 34380

gaatgtcatg tggcatcagg tagcatacct gtatgagcag tgcctcaagt acatgaagct 34440

tgaggttccc aactgcatgc agcgtaacaa tttcatccgt tacggcttac aggagtccgc 34500

cttcattgag aagcgtaact ccaagcggta caacggaaga tacgtcatcc gcgagggtgg 34560

caaattgcgg caactgattt gtatcaagct gccgcataag tccggtatca aagaagactg 34620

gactaaagac gagatgaggg gcatcgtcct ccgctcggtt caaccgctgt tgaatccccc 34680

tcgtggatca cagtaatcca atggcttcgt agagaagcttttccacatca agagagatct 34740

ttcatgacta tcaagtccgt acaactgttc gttaccaccg atggcaagca attcgaagac 34800

caggctaccg ccgaagcgca ccaggtttcc ctcgacaacg ctgccgtagt tggcaaagtt 34860

gccacctcct tcgtcaacgt cgccgttgct ccgaacgcca aggttcccgg cctgaacggc 34920

cgcacccgtg tgttcaacca gaacgtcgcc agccaggttg tatccttcct gatcgcccaa 34980

ggcgtgatcg ttgctgccga cctggaagcc ttcgaggcca tcgagccgag cgaagagctg 35040

gctgcccgcc tggaagctga ccgcctcgaa gccgagaaga aggctgccga agccaaggcc 35100

aaagctccct ccaaaggtga aaccgctccg gccaccaccg gtgaagaaac caaagaagaa 35160

gtcgctgaag acctgttcta attctgatcc ggcgcgctgg catcgcggcc agtagtcatc 35220

gactacaggt aactgccaaa tctagcccct tgctgaaatg acgcgagggg ctattttttc 35280

ccaccgacca tcgagtcatt cagaatgcac tcaatggtga gtgtattccc aatgatacgg 35340

agattccaat ggctaagccc tcgactaagt atccccgtcc tgctgagatc ctgatggctc 35400

tggcggcggc taatatcgac caccggttca tcaccgtacc caatccgcag cacatcgaac 35460

aggagttggc acggctgatt cccgacccgc gtattgctct gcaagtgtct cgcctgatgc 35520

aagagaacca gaacggcttc atccaggcgc tgctgcacac cggcaaccag gccctccaag 35580

ccatgggcta ccaagagacc gacgtcacca aagacgctgg tggtgtagcc ggtcgtgagc 35640

tggttgagca agtaggacgt gctatgcacg ctgctgctga gctgaagaag gctgaagagg 35700

ctgctaaagc gcctgctgcc aaggagtaat catggctggc acttgcgtag agcggatgaa 35760

acatgatgac tcgaagtgca acagcacctc gaagtcgctc cagatattcc tcaacgatga 35820

tgataccttc tcggggttct gtttccactg tcacaagctg gtgcctaacc cctatggtga 35880

caatcctcca gacccgaaag atatccatgt caagacaccg gaagaaatcg cggaagagat 35940

tgctgaggtc aggcagtgca aaacaatgcc tctcaagcat cgcggcatcg acccagaaga 36000

ctggagttac ttcggtgtcc ggctcctcac gagcacaact gacggcacta caccgtatgc 36060

tatcgcgcat ccttatacca agaatgcaaa ggtagtaggc ttcaagatca agctgatcgc 36120

acgcaagacc atgtggaacg tgggtgacgt taaaggcgcc gacctgtatg gttgggaacg 36180

ggcgaagcgt gtaggcggtg gtgtactcta catcaccgag ggtgaagagg atgcaattgc 36240

gctccgcaaa atcctccgca tgacctccac caatagctcc tacgactacc ctgtggtgtc 36300

gttgcctgcc ggtaccaatt ctgtggccat gtgtctggga cgtatgtccg aggaaatcaa 36360

cacacgcttc aagcgtatcg tgctggtgta cgacgacgat gagcccggcc gtaaggctgt 36420

gcaagaaact cgcaagatct tcccgcatgc tgagtctgct atgctcccag agaaggacgc 36480

caacgagtgt ctgatcaagg ggcgtatcaa ggccgccaag gatgctgtag tgttccagat 36540

ggctaagccg tcacaggcgg ctatccaggt gctcagtgcc gatgacgtca tggacgagat 36600

tatgcaagac ccagagtggg gcttgtctta tccgtgggac gacctgatga agaccacgta 36660

cggccagcgg aagaaagagc tcatctccat cggcggcggt actggttgtg gtaagaccct 36720

gattggccat gagatggctg cgtggaacgc tcgtactcac gggtggcgca ccctgatgat 36780

catgatggaa gagtcaccag cagagacgtt taagaacgtc gcgggtaaga tcgacaatgt 36840

gccgtaccac atccctgtca aagatggcga agagccatac gacaaggaaa ggctgcggtc 36900

caccatcgac tacctcaagg ctttcataag cacctgggat atctcgacca ttgaagaccc 36960

cgagaccact tgggctcaaa tcactcaggt catccggact caaggccacc tgtacgactg 37020

tatcatggtg gacaatgcta caacgctgtc ggaaggtctg tcggcatcag agcgtaacga 37080

cttcctgggt aaggtgaaca acgagtttgc caagctcgcg gagaagtttg acttcgaagc 37140

aatcatgttt agccacctga atgcaccgcc gtccaaccaa cgttcccacg agaacggtgg 37200

taaggtaacg gaagcccagt tcactggctc tcgtgcagcg atgcgttact cacacatgat 37260

cttcggcttc gagcggaaca agtctgcgga agacccagac tgctcatact tcgtggtcct 37320

caagaaccgg aagttcggcc gtactggtcg cttcaagacg tactactcac aacgtactgg 37380

tcgcctgcaa cagcgcaact gggacgatga gtcgtaccaa gacaaggcta cggcaagcac 37440

caaagcctcg taaggagaat ccatgctggt agtacctgat gcgtgggaac tgattctcaa 37500

gactcagaaa gacgggtctg agaagactgt taccctgcaa aagtttgtag aagtaactgt 37560

ccctcgcaat agctactccg ctaaagcagt ggacaacctg atggctgttc gtgagcgact 37620

caagaacggc gaagaagtag atggtgccga gatcaacctg aagggagtga ccttccggag 37680

ggcaccatga ccgaccaaga attcttccga cgttctgtgc tgatgttcaa gagcacatac 37740

cctgaaatcg gggacaagac agccattgca cgcgcgcaag agctgactga cgacgtagtc 37800

gaacatgagc agaaaagcgc catggctcgc tacaaggctc cggaaaagta cgagcggatg 37860

ggcaagggca gcatcgtaaa gaaggcctgc gggttctgca tcggcaccgg tgaactgcga 37920

ggtggtagcc aatcgttcgc ctgcccgcattgcaatggta cggggaaaga gcaatgactc 37980

atcccatgta cctgctctcc gagccgaaga acatgttcgc ggcagacatt gaaaccactg 38040

gcctgctgga agacctctat acccaagagg caccgaagct ccataacttt ggtgccaagg 38100

ctatggatgg tactgagttc ctgttctccg aggcctggaa cgtcctgaac cacaaagcct 38160

gcgcggacat ccgaccgctg tcagagctcc aagcattcct ggatactggc ccttgcctca 38220

tcatgcacaa cggtatgtgc tacgatggtg aggcattgat cttcttcggg tacgacatct 38280

ctaaggttcg tattctggat acgctctacc tctcttggta ccttgagccg ctgcgtaatc 38340

gctacggcct tgccgagtac ggggaagaat ttggtatacc gaagccggtc attgacgact 38400

ggcagaacct ctcccaagaa gaatacaacc gacgggtcat gcaagactgt cggattcaac 38460

tccaactgtg gaagcgtctg cgtgctatgc tgtatcgtct ctatgatgat attgaagctg 38520

aggcatggaa ggcgctgaac cacgtgtgtg tgataaaggg ccgccatttg cggaaccaac 38580

agcgcacaaa atggaccgta aatattcctg ctgcaaaaga gctgtccgag cgactgcacg 38640

cagacaagga agaccggttc gtggagctga tcgaggttat gcctcgtaag ccagtgttca 38700

agagcaagaa gcgctgcgcc aaaccatgga agatgaacgg cgcactgtca tctcacggtg 38760

aaggctggtt gaagttctgt atgctgcacg gcatcgactt caatgacctg accgagttcg 38820

agtaccaaga cggggacgtc gagggcaacc cgaacgctcc acagcaggtc aaggactggc 38880

tgttcagtct cggctgggaa ccgcagacct tcgattacaa gaaggatgat gctggtaacg 38940

agcgagtgat accccaaatc aacgtcaaga atactggcgg ggagatggac ccaggtatcg 39000

agttgcttat cgaggaccat ccagaagttg caagactgaa gggtcttggt atcgttaagc 39060

accgtatcgg cattgttgac ggctggattc gtgacgccat tgatggcaag gtcatggcac 39120

gtgctgctgg cttcactaac actctgcgtc tcaggcacgc tgaggtggtg aacgtcccgt 39180

cagaacgggt accctacggc aaggagctgc gctctctgct gatggcgcct gatgacaacc 39240

atgagctact aggctctgac ttgtcgtcgc tggaagaccg gtgtaagcac cactaccaga 39300

tcccgtacga ccctgagtac gtgaagagcc aacaaactcg tggttacgac cctcacctgc 39360

gtatcgcacg tctgggtggt ctgtgctccg aggatgacga gaagttctac attgactaca 39420

cgcttcgtga agaggatggc ctgccaatta ccgaggaaga caccaagcgg ttcaagccgc 39480

tcaagggtat ccgaggtaag ggtaagccaa tcaactatgg atgccaatac ggccagcggc 39540

cagcaggtat atctcgtgca tctaagatgc cgattgagat ggctgagctg ctgtacaatg 39600

catactggga acttaactgg tccatcaatg cagtggcgga ctccatggag atcaagtcgt 39660

gtaacggtgt gaactggcag cggaaccctg tgtcagggat ctggtaccac ctcaagacca 39720

acaaagaccg gttctcaaca ctgtgccaag gtactggtgc gtacgcattc gacctgtggg 39780

ttgagtctat cttctgcatc tgcgaagaac gctggaagcg tgagcctctg ctgaacggcc 39840

agttccacga tgaactgatc ctgacagtca agaagggcct gcaagacctg tggcgtggtg 39900

tagtccgtga gggtatcacc agagcgaacg gcctgctgaa gatgcgtcgt gaaattgatt 39960

gcgatatcca gtttgatact gtgtacgcgg gaatccactg atgtctcgat tcgaaaagat 40020

ggtcgccggt aacaaggcga tcttcgaata ctgcgaacgt attcgacaaa tgaaccaaca 40080

agaggagaag cctaatggct ttgtttgata aagcaactga gaagtctgct ggtggtaacg 40140

gccagaagat ggaagccccg ctgctggcgc tgggtggcta ccctgccgta ctgatccgta 40200

tcgtcgacct cggcgagcag cccggctccg ctcagtaccc cgacccgtcc tacaagatgg 40260

cgctggtatt cgagtgcctg gacgagttca tggtggacga agaaggcaaa tccctgccgg 40320

acgttccgcg tgagttcgac atggaagtca gctacaacct tgacggctac atgtcgccga 40380

aggccaagct gcacggtgtc atgaccgcgc tggaaggctt cgacaagtcc ctgagcgagt 40440

tgctgggcac tgtctgcacc atcaacctga tccagaaggc caccaaggcc gacgcgacca 40500

agaagtacaa cgccatctcc ggtgttgctg cgatgcgtga gaaggacaag gaacgttacc 40560

ccggcccgtt caagctggag cagtgggtgt tcaacctaga cgccaacacc accaaggaag 40620

acttcgagaa gcagtcctct cgcggtggtc agtacagcca gcagtccaag atcaagggcg 40680

cactgtccct gcacaaagac gctccgcaac tcgcggccgc tctgggtgtg gaagctccta 40740

agatgatcga ccacggtgcc gacaagccgg taggcgaagc cggtgctgcc ttcgacgaag 40800

aagagcagaa caagcagatc gaagctgcta tgggtggcgc agcagtgcct acggaaggcg 40860

ctggtgcgga accgaacccg ttcgattaat cggagacgac atttatggca attccaacaa 40920

tgtcgaaatc cgttgtcgac gagcccgaag acggtcgcta cgactacgaa gagctcctga 40980

tcgacggtga cttgttggtg ttttcctcct gtgcggctgt tgaatacggc cgcactccgg 41040

aagaatacac tctgcaagaa atcctgacca acatcgaagg tcggatcatg gcgatgaagc 41100

gtcgcctcaa ggctcgtaaa gtacgtatct tcttcacggc ggaagacaac ttccgttaca 41160

agatcatgaa ggcgtacaag gccaaccgtg aaggcgcgtg gctgccggaa tccttgaaga 41220

atgcaaaggc ccatatcacc accatgttca atggcgagcg tgaaccaggt ctggaagcgg 41280

atgacctcat gggcatctac cagaagattg atggcaccac catcgttgca accatcgaca 41340

aggacatccc gcagattccg ggcatgcact accgctggga gacccagcac aaaggcgagg 41400

ccgtctttga ggtgtctggc tacggtactc tcgtgaaaga aatccacaac aagaagacca 41460

agatcagtgg cacgggtgct cggtttttct gttaccagct gctcatcggt gatccgactg 41520

acggcgtaat gggttgtggt aagctggaag agtcggtcta caagactggc gcgaaggctg 41580

gcgagagcta caccaagcgt tccggtgtcg gccctgtcgg tgcctacgac ctcctggagc 41640

atgcgatcac gtatcaacgc tgcatggaga tcgtcattgg tcagtacaag catacctttg 41700

gtgacgcttg ggaagaacaa cttctcgaga atggccggtg cctgtacatg acccgtaaga 41760

tcaacgacaa aggtcaattc cagctgtggc atttccgtgc cgaagagttg cctaactctt 41820

ggtacgaccc tgctactgct gctgtagtac gtttcgaagg ctagttaata tgagcgcata 41880

cggttactgc aaaagcgaga aagcttatgt cgcttggttg agatctgcac tcagacgagt 41940

gtggtcgaaa cacccagtga agctagagat gctcaagaag aatcgtgtgc gctctgttaa 42000

tccgaagaca agtcgaatgt gctttatgat cacgtgcaag acgtgtggta aagaccatcc 42060

gttgtcggat atcgaagtga accacaagaa tactgtggga actctatcac tggagaactt 42120

tggcatttat tgcgaaagac tcctgctggt agaggagaaa gacctagaac ttctgtgcaa 42180

gacctgccac gaagttgtca cctatcaaga aagatctggg atgacactcg aggaagctgc 42240

gattgagaag agagttattg ctttcttcaa caagtatcca gcaggtgaac agaagagacg 42300

gttcgaattg ggaggcatgg tgcctgccaa gaccgtcgcc gagagaagaa ttcaactgag 42360

ggagtacctt cgtgaacaag ctagaaatgc tggaactggc ggctgagaac gccagataca 42420

acaacgctgg gaagaagtgg cggaagatca gacatccgaa tcaccctgac gaccattaca 42480

caccacatct gatactgcgc atgaagcaga tggatgggtc ctggacaccg ggaatcgtgt 42540

acattaacag cgctggcgaa acattctgcc gtgctgctga caacttcgag aagttctcgg 42600

aggtgtaaga tgcatgagac cattcaaacc atgaacgcat tgcgtagcgg gatggttatg 42660

cgttatcacg cgcagccaga tgtaccgtgc cagtccaacg ccgagcacat gtggggcgtt 42720

gcagtactga tgctgaagtt ctacggcata gaaatgagcg gagaagctct tgcagcagcc 42780

ctcactcatg actgtggtga agccgatgtt ggtgacattc catcaccgac aaagtacaag 42840

attccggaaa tccgtgatct gatcaagcgg ctggaacacg agtctctcaa tgaactcggt 42900

ttcaacttcg aagctgacct ccatcagtca gagaagcaag cccttaagat ctgcgatgtg 42960

ctggaggggt tgcactacac cgcgaagcat taccacctga ctggtggtgc tcagggtggt 43020

caatgcctcc agaactgggt ggacctagcg aagacactgc cgctgacaag gccgcaacgt 43080

gacttcatct acgtgtgcct ggcaggcccg tcacgcagag tagtatcata gtagtgatca 43140

ccgacgtgtt ctaccaaggc gcagatgatg gcatcccagg ccactggtgt gctcagtact 43200

tcaaaggagg ctgggcgcag taccaccgca gcacggacta cgacgagctg gaacaatgga 43260

tcaaggagaa cgccaccaat gcggatgcct ggatttaaag tacaccaaga agtgaagaac 43320

acgttgtgga ttgctgcgat tgtggcaggc atagtgttcc actgctgggc aatggcgaac 43380

agcggagtac tactctgctg gttctaccaa gctaaactga gagaattctg atgcaacaaa 43440

gctcactgag tatctccatc cagaaagcac tccatctgca tctggaagaa gagaagaaat 43500

ccccgaagca cctcggtcac gagtacgcga tcttcgagaa gtacgatggt tggtacatgt 43560

acgttgactg tatcgacggc gtgtggcaag gtatccgctc caagacaggc cgccttctgc 43620

cctccatgca gtggtacaac aacctgctcg aacaaggccc gaagcctaat caagacctgc 43680

gcctgatctt tgaagcagtt atccctggga tgatcttcaa ggacgccaac ggccgcttca 43740

accagaccaa ggagcagctt ggtaacgtag tctgctggtg ccatgatgtg ctgtttaaga 43800

aatacccgca caccatcttc tccaagcgct acgaccgtct catctccgtg ctgaaccacc 43860

tggactatag ctggctgcgc cgtgcgccga ttctggctgt gacccgtgac gttgccgtat 43920

ggcatgagca ctacgagcgt atcatctcca ctccggacgc gtacggccaa aaaggcgaag 43980

gcgttatcgg gaagaagtgg gacgagccgt accacgaagg caagcgtgac tactccttga 44040

tgaagatcaa gtgcgagctc actctggacg ttgaggtgat cgctgtcggc cctggtgcga 44100

aaggctccaa gtacgaaggc agccttggct tcatcgtggt tcgtgagaag agcggccaga 44160

ctcaccaggt atctggtatg tcggatgctg aacgagttga gtggtggaac aatccccagc 44220

tcatcgttcg caaggttgtt gaaatccaag ccatgaagcg cttgccgaac gggtcactcc 44280

gcgaagctcg ttacaaggcc gtccgccacg acaagactct ggcagacatc gattaaggag 44340

aaccgaatgc aaagagtacc tactgtagga gtggccttca acggccctcc tggcattggc 44400

aaagacacgc tggtgaagct catcaacgag gtactgttcg acaagattcc tgtagggacg 44460

cttgccaaca tgatccgtca ggatgcagct gcgtactacg gcatcgagaa cttcttcgag 44520

ctgtcgacgg accgtgagac caaggacacc aagtgcccag gtttcagcat gacccctagg 44580

cagctgctga ttgattacag tgagaatgtg atcaagaaga atcacggcaa ggatttcttt 44640

gccaagaagt tcgccgagca gatgtctcaa gagcaacagt tcatgatgac cgaccttggc 44700

ttcgttgaag aagctgaagc gctggccgac cgtgtcgacc tgcttatcat cgtgcagctg 44760

gagcatccgg acttcgactt cagccgtgac agccgtgggt acgtgaagct gcatcgcagc 44820

aacgtattca acctgccttt cgtcgtgtct cgtggcgatg cccaatctga cgcccgtcgg 44880

attctcgaag ctatccaaca tgcttatgag gaacgactcg ggttgtaacc aatcaggagt 44940

ccgagttgga ccttagcaaa tacatgaagc ccgttgacat tcgtgtcgta gtaaaaaggg 45000

atggcagcga ggagcctttt gactcctcca aaatcgagcg ttgggcagag tatgcctgca 45060

agcagaaagt gaactggaaa ggcgtagcgg tcaacgtgat ctctcgtctc ccgtccaagg 45120

tcaccacggc tgagattcat cagtcgatga ttgactactg cctggcgcaa gaatctctgc 45180

cgtacagccg tgtggctgct cagctggtat atgctcagct gcgtaagaac atgcgccgcc 45240

acctgagggt tgatgaccgt gacgatatcg gtgacatcgc cttccaactg gcggagctgg 45300

gtctgtggac tcatgagctg tccaatctga tcactgacaa cgaagagctg gtgcagggat 45360

ggtacgacgc catataccct gcgcatctcg agtcctggca agtaatgcaa tgggcggaca 45420

aatatgcgat caagaaagac ggtattccgg ttgaaacacc gcacgtgggc gcgcttggaa 45480

ttgcaattgc aatccatgga gtcacccaag atgcgtttcg actggctcgt gccatcgtgg 45540

aaggtaagat taatctgccc acgcctgtac tcaatggctg tcgtaacgga gactttgact 45600

caatctcgtg ctcggtcatt actggcggcg acaccgttga ctccatcggt gtggcggagc 45660

accttgcgta caagatgact gccaagaagg ctggcatcgg tatcgagatg aacactcgat 45720

ctgccggtga caaggtgaaa ggcgggcgta tcaagcatct gggtaaacac ccgatttacg 45780

ctgccgtcga caagtctgtg aagatgttca cccaagtcag tcgtggtggc tctgcaacca 45840

tgaccttctc ggtgtacgac cctgagatta tgatgctgtt gaagctcaag tcacagcgta 45900

ctcctttgga tgtccgtctg gataagatgg attactcaat ggcctatgac gataacttcc 45960

tcgaagcggt cgtcaagaac actgacgtcc agaccaagag cgttgacggt ggtctcgggc 46020

cggtctaccc tgctcgggaa atcctcaagg tgttcctgca aatgcgtcag gagactggcc 46080

ggctgtactg cttcaaccac tccgaagcga accgccacac gccgttcctg gacgagattc 46140

gcctgagcaa cctctgccaa gagatctgcc tgccgactgc cccgttcacc gacatgctgg 46200

acctttacaa tccggcactg ggctacaagt ccaatggtga gattgcattc tgctctctgg 46260

ctgccatcaa cgtgtacaag gtctccgaag ctgaatatgc tgacatcgcc tatctggcgg 46320

tgaagaccgt ggacaacctc atcacgctgg cccccgctct gagtgcccca gtggccgaga 46380

aactgcaagc gcggcgttcc ctaggcatcg gcattaccgg cctggcaggc tggctaggcg 46440

agcactctct gcgatacacg gatacggcct cgattgagtc acttgcagaa cgccactact 46500

actggtgcct gagtgcctcc caacgcctcg ttctggaagg cagagcgccc gtctctggca 46560

tccgtgagga ctggctaccc atcgatacgg gccacatgac cgttgcgcct cgcatggact 46620

gggaagcact ccgtggcaga cccagacgga actctgtact ggtagcccac atgccgaccg 46680

agtccagcgc cgtgttcagt gacgccccga acggcctcta ccctgtgcgt gacgccgtca 46740

tagcgaaggc ttcgcggtac gggcttatca agtacatcgc cccagccgct tgcacagaga 46800

gagcgtggga cgtgggcaat gatgtcctag cccgtgcgta tggtgccgtt caggcatata 46860

ccgaccaggg catctctgcc gactactacg tcaccccgag caagtacccg aacgggaaag 46920

tgcccatgtc ggtgatgatc aaggagtggg ttattcaagc caaggctggc gtgaagacca 46980

tgtactattc caacaccaac gacttcaacg gcggtgcgtt ctcccagcaa gaagatggct 47040

gcgagggtgc ctgcaagctg taaggaagaa tcccatgcaa attgaattga cagtggacgg 47100

gaagtccttc aaagtagagc ccatgcaggt agaaacccaa gaggagcgat ttgactcgac 47160

tgtcaagtcg tatctgctgg ctgctgcaca gcaaatcatt cggaagtatc tgttagacca 47220

acaataccct atccggtaca accaacctga agtacgcaat ctggaaaaca ttgtggactc 47280

agcgttcctg cgcagtggcg ttcacgtcgc ttcgaccatc accagcaacg ataacgggag 47340

aaccatgaat gtcagtcttc aacgccgcta ataccggtta catcaacggc tatccgaaac 47400

tgttcctcgg cgaacagcgt ggcctgctgg acactatcaa tgtccagtac cctgagctgg 47460

aagctcttta ccaacagcag atgtcccaaa tctggaacga gttcgaagtg gacctgagcc 47520

aggatcgcat ggacatgcaa actgctgctc cgggtcttat cgacctgatg gtaaagacta 47580

tcagctggca gcacctcgcc gactcggtag ccgcgcattc catttccggt ttgctgatgc 47640

cgcatgccac caactccgag atggagagcc tcatcaacgc ttggtcgttg ttcgagacca 47700

tccacgctcg tacctacagt cacatcgtca agcagacctt cgcggatcct ggcaagatgc 47760

tggaagagac ttacgccaat atggacgtac tgactcgttc caacgtgatt gtagacgcct 47820

tccatcgtct ggaagacgcc ggtgcaaaag cagaagaggc cgatatcatt atcgctctga 47880

ctgcgttgtt cgctctggaa gagattggct tcatggcctc atttgctatc actttcggca 47940

tcgccgagaa tgcgaagaag ttcatgggta tcggccagct ggtgaagctg atttgtcgtg 48000

acgaagtcct gcacactcgg tttggcggtg cgattctcaa gatccaccag aacattccga 48060

gcacctctgc gctcatgcag aagggctggt tccgtgacga gctgaaaggt atcctggatg 48120

ctatcgtcaa ccaggagctg accttcaccg actacctctt ctctgagggt cgtgaatgca 48180

ttggcatcaa tgccgatctg gtgaagcagt acgtactgta tatggccaaa cccatgtaca 48240

tcgcgtacgg cctgaccttc gactttgatg ctccgaaaga aaatccgctg ccctacatgg 48300

accactggat tgacggtagc aaagtgcagg tagctccgca agagttacag aacggttcct 48360

accaagttgg cgccatcgtt gatgacactg acgatctgga cctggatgat ttgaacctgg 48420

gttaatcgtg gcaaaagaaa atgtcttcga gccagtagat gagttcgagg acaacttctt 48480

agtcggctgt atgctgcaga actatggcga cctgttaaag gtctccagga cgaagggtgt 48540

gttacgcacc cttccgaatc tgcgggaata cattaaagac aaccagtaca ttcgttacga 48600

atacctggtg gcgcttgaag cagagcgtga tggtcaaggc ctgactggga acgctgttct 48660

caattcactg atcactgcgc agctggctgc atttgaaagc ggtgaccaca agaacaccaa 48720

gatgtactcc gatgagatca agaagcttat cacggagggc atcaagacat ctaccgttgc 48780

tgaggcagtc caacgagcaa tcgctgggaa gccttcgctg gtagaagaaa tcctcaccag 48840

gccgttaagc gtcgaccttg acgcgatcac ggttgaggat gcaaaccgag tacgagagat 48900

cttacttgat cccaacaatg ggttcgagaa gttcagtaag tggtgctttg aaatccagat 48960

gggtttcaag tttcagatgc aggattttca ctcgattata tttgagttct gtcagaagat 49020

cgtcaatggc gaaatcgaca gaggcattgt ctgcatccca cctcgtcact cgaagactca 49080

gatactgagt atcttccttc cgctgtactc gttctgcaat aacccctcca gtcacaatat 49140

aatcacttcg tacgcggacg atgtggtggc ggagtcctcg ggttatatcc ggcagataat 49200

gctggactcg ttgttcatga agatcttccc tgctgtacgt atcgaccaga acaaacgctc 49260

ccttgagcgc tggggtacta cacggtctgg cgtaatgcac gctgtaccaa caggcggtaa 49320

gatgactggt aagggcgctg gctcactgtc accaacctat tcaggatgct tcgtggtcga 49380

cgacgcgatc aaaccgaagg acgcatactc tgcgcctgtt cggaaagaga tcaacgaccg 49440

attcgacaac accttcatgt cgcggcttgc gaacgacggt gaagtccagg acgatgacgg 49500

taatgtcgtc aagtgtccaa gaacacccat ggttatcata atgcagcgag tgcatgatga 49560

cgacctcgtg gggcacatcc ttcgtggtgg ctcctcggat aagtacacat acctgaacat 49620

tccaggtatc gtgacgcctg agtgtggtac cgagaagtgg tacgagaagc tgctgacgaa 49680

gcaagcctac actcacgcgg ttccgtacct ctacgacctg aaacgcggcg aaggtgaatc 49740

tgcgctgtgg ccaagtcgta agagcctcga ttctcttatt gcgatgagaa gcgcgactcc 49800

gtacacattc aactcccagt acatgggtga cccaactgcc agaggcacag gcctgatagt 49860

agaagactgg tggcaggaat ggacggagct tccgttagat gatatcgctc gcgcattcat 49920

gacggcagat acagcctcaa ccaagcagga ctactcagat tactccgtga tctgttattg 49980

gctcctgact aagaaaggca atctgtactt agcggatgtt gagctgggga aatacgagac 50040

accagagttg atgaaagtag tagtcgactt ctggaagaaa cacacgcaat tggatctacg 50100

gtatcctatg ctgctcccga cggcgctgta catggaagat aagtcctctg ggcaattcct 50160

gaaccagcag ttcacgagag acgggcaagt gcgtgttctc ccagttccga aagataaatc 50220

ttcgggtgat aagattgctc gattcctaaa cactgtgccg tacttcgcac agggcaggat 50280

ttatttccct gcagaacaca aacacaaagc acacataatg cgagaaatcc ttggtatgac 50340

tggggaaggt tctggtacag atcacgacga cgttgtagat aacgtgtctg acgccgtagc 50400

cgtagcattc agtgcccaga ctgctaacta tgcagcctgg atgtaaggag aagctaatgg 50460

gtatcaagac ccgcctggac caacggtctg aaggcaacaa ccgtatgatg ctggtcgatt 50520

ccgccactgg tgaaatcgta gccatcatct ccgcagtgtc caacaaagtc gagatcgaag 50580

tcgagaccgc gaagggcctg cacatcgaga agccgaacgg gttctccagc aaacgctaag 50640

gacaatcaat gtctgaacct accaatgcac tggcgaagcg tgaggaagac ttccgcgctc 50700

gtctgatgat gcatgacggt ctagagaact tcatgaccgg tctcggcact ggcaaagaca 50760

agaactcgta caacgagtgg atcaggtctg gccggaaccg tgatcatgaa caactcatcg 50820

tcaggtacag ggaagactgg gttgctcaga aagtttgcaa catcctgccg gaagacatga 50880

ctcgggagtg gcgccgatgc agtacacccg aagccatcga ggctgatgat gaattccata 50940

tcagccaact cttccgtgag gcgtacactt gggctagagt gttcggtact gctgctatta 51000

tccttgacct taagggtact ggccgtgcag agacaccgct cgacctgagc cgcctcaagc 51060

cgggatgcat cagaagtctc caggtagttg atcgtacacg gctgatgccg tcaggtacgg 51120

ttgaaatgaa cccgatggat ccggagtacg gctacccgca gcattacatg ctgggcggct 51180

ccacactgca aatccaccgg actcgtattc tgcgctttga agggactccc ctcaccagat 51240

acgagaactg gaagaaccag tggtactcag actcaaccct gatcccgctg tgtgagacca 51300

tcgacaactt ccacacggct gcgcaatccg cgtctgcctt ggttaacgaa gcaaacgtcg 51360

atgtggtcac tgtccaaggg ctgcagaatc tgttgaccaa ccctgctggc gaggctgctt 51420

tgtataagcg cttccgcatg atgaagcaga tgaagtccaa ccacaacatt ctgctgctgg 51480

acaacaccga agagtacagc cccaagacca tcgcactgaa tggcgtcaag gatctcatct 51540

gggaatactt acgtatcatt gctgctgctt gtggtgtacc tgcaacgaga ttcctctcgg 51600

cgtcgcccga tggcatgaac gcaactggcg agtcagacct caataactat attgacctgc 51660

tgcgccagaa acagaagtct gtctttgaca aacgactgcg tgtcatcgat aagattctcg 51720

ctgctcactt cggcatccag ccgttcaagt atacctggaa ctgcgcattc ccagagtcgt 51780

ccctgcagaa ggaagaacgc cggaacaaac tggccgattc attgcagaag cttgtagttg 51840

gtggcgtcat tactggcgat actgctatca agattatgga agaagagcat actttcggag 51900

acattgatct cgggaaggca ccgccacctc ctcctaaacc gtcaaatggg gaaaactcca 51960

aatgatcaaa atgtctcaac ggatcacgtt ccgagacgaa gctgagcttg agctgacccg 52020

ctttgttgat gaaggctgtg accttccaac tcagcgcaag ttcaaggata gcggtgcgat 52080

gattgctccg gcgaccattg cccgcaccgg catcatgaac tactctgctg gccactgcgg 52140

caagttgttc tccgactggc ctgcgaccaa gatcgtgcga gtgatgactc gtgccgaaga 52200

cctgttccac gccgactccc tcgaactgta ccgttctgcc ccgatcacta tcggccaccc 52260

tgacgaagac gtgtccatcg acaacgccgg ctggctgcag aagggtaacc tcgatggtat 52320

tcctttccag gatggtgagc agctcgccgg tcacgtagtg ctgacccaca aggaagctct 52380

ggacctggtg gatgctggtg taagccaact gtcctccggt cacgacgcta ccctgatccg 52440

cctgtctgac gaagatgctg cgcgactggg ctacgacgcc tacaagacca acatccgtcc 52500

gaaccacgta gccatcgttc cgaaaggccg tgccggctct gcccgcatcg ctgacgaaga 52560

cgaaggcgct ggcaagacca aggaagacga aggtcaggtc aagatgtacg accaggccca 52620

cgtgtccggt ctggaagctc agctggaagc atctgtcact ctggcagacg gcctgaaagc 52680

cgaggtgcaa gagctcaagc tcaagatcac cgacgaagcc atccaggtgt tggtcaacaa 52740

gcgtctggag ttcatgaccg aggtggctca gtttactgat gctgatatct ccggcatgtc 52800

cgagctggat gcgaagaaga ttgctatcaa ggatgcctgt ggtaaagact acagcgaccg 52860

tgacgagcac ttcatcaacg cccgctacag cattctcctg gaagaaggca ccgaatccgg 52920

tggcaccgat atctcccagg ccttccgtga tcacattaag gatccggaaa tcaaagcgaa 52980

gtccgagcct acgcagagcg aatccgcccg ccagcgtatg atcgaccgtc aatccaagta 53040

atcgaggaca caacatgccc gttcaagaat atcacatcaa tacccgtgac gaagtcgaag 53100

gtcaactcta cggcctgcaa cagactcgtg ccgacatcca gactggtttt gccgaagatg 53160

tcagcggcat cccgttcggt aaggccgtta aagtcggtgc tggcccgcgt ggcttcctgc 53220

tgggtgccgc tggcgctcac gtagacggca tcgtcctgcg tcagatcgac cgtgaagctg 53280

ccaagcgccc gtccgacggc accgtagtct tccgcaaggg cgactccctg ccggtgtgct 53340

ccgatggccg tgtggtcgtc aaggtgatgg acgctggtgc catcgttcgt ggtcagaagg 53400

tattcgtgca tcaagccaac ggtaccttcc acgtagctac tggtgctggc ctggtcgaag 53460

cctccaacgt tgtttggggc atcggtaaga ccgccgctgt cggcgacctg gtccacgttg 53520

taatcaccaa cgctgacgta gcctaattgg ctgagccgga gcaatccggc tcttctccct 53580

cgattcaagg aatacaaatg tcccgtaaag taaagttctt cgacgcagag caaggcaaag 53640

aagtagagat cgaactgcgt gctgatatcg ctcgcctgat cgaccaagat gttcgtctga 53700

ccgatgacga cggcatcttc ttccagcgtc aactggaagc cattgaagct cagacctacg 53760

atgtgctcta tccggacctg gaagctcgta gctgcttcaa aaccaacacc ttcggtggtg 53820

ctggcgcccg taccctgacc taccgttctt tcgaccgtat cggtaaggcc caggtcatca 53880

acgctcgcgc taccgacctg ccgaagtccg acatctccgg taaagaatac tccatcactg 53940

tcaagtccgt aggcgtcgct tacgactacg acatcgacga gatcgctgcc gccaagatgg 54000

ctggtatgcc gctggaagct cgtaagacca tggctgctcg ccgtggctac gaggagtaca 54060

tcaactccgt tgtatggcgt ggtgatgctg atgccggtct gggtggcctg ttcaccaaca 54120

ccgacatccg ccgtaccgct gttgcccaag gcactgctgg taccaccggc tggagcacca 54180

agactccgga cgaagtgatc gctgacctga cccacgcttg tggcgacatg tacgcgacca 54240

ccaagaagat ccatgctccg aaagagatct ggatgtcgac cctgaactgg aattacctgt 54300

tctccactcc gcgtagcccg atgtctgaca agaccatcgg tgactacttc tgcgagaaca 54360

accagttcgg catcaagaag gagaacatca agccgctgaa cgagctggct gaaggtatcc 54420

ccatgggcac cggcgcaggc tctcactgtt t 54451