CN112662634A - Drug-resistant Klebsiella pneumoniae RDP-KP-20004 and application thereof - Google Patents

Abstract

The invention discloses a drug-resistant Klebsiella pneumoniae RDP-KP-20004 and application thereof, wherein the preservation number of the phage is CGMCC No.21407, and the whole gene is sequenced and then is found to have no virulence gene and lysogenic gene, and the drug-resistant Klebsiella pneumoniae RDP-KP-20004 provided by the invention has higher temperature tolerance and wider acid-base tolerance range, has strong cracking effect on pathogenic Klebsiella pneumoniae BK-20004 in a poultry breeding environment, and provides a phage source for the industrial production of phage for the prevention and treatment of the pathogenic Klebsiella pneumoniae BK-20004 in the poultry breeding environment; the Klebsiella pneumoniae BK-20004 provided by the invention has good advantages in treating diseases infected with Klebsiella pneumoniae, and has high titer and good prevention and treatment effects; has good application prospect in preventing and controlling the pathogenic proteus of poultry.

Description

Drug-resistant Klebsiella pneumoniae RDP-KP-20004 and application thereof

Technical Field

The invention relates to the technical field of biology, in particular to a drug-resistant Klebsiella pneumoniae RDP-KP-20004 and application thereof in pathogenic Klebsiella pneumoniae infection of poultry animals.

Background

The discovery and application of antibiotics are a great revolution of human beings. Has played an important role since the world, has special curative effect on bacterial infection, and promotes the rapid leap of the medical level of human beings. However, the irregular use of antibiotics, resulting in bacterial resistance and even multidrug resistance, is becoming an increasingly prominent problem. Medical researchers have indicated that approximately more than 50% of the antibiotics are abused worldwide each year, and that this proportion is even close to 80% in china. Besides the medical field, the situation of overuse of antibiotics in the planting and breeding industries is also serious, which directly affects the food safety and threatens the health of the whole human.

Bacteriophages, also known as bacterioviruses, are microorganisms that proliferate in the body of bacteria, releasing themselves into the environment by lysing the bacteria. Phages were discovered and attracted extensive attention in 1896, but with the subsequent discovery and application of antibiotics, phage research was in a state of stasis. Until the 80 s of the 20 th century, researchers began to focus on the study of bacteriophages due to the emergence of multiple drug-resistant bacteria. Compared with antibiotic treatment, the phage treatment has the advantages of low metabolic efficiency, low minimum dosage for killing bacteria, high specificity, high efficient sterilization effect, high screening speed and the like. In recent years, the research on bacteriophage has been greatly developed, and the application research range of bacteriophage is wide at home and abroad, including the aspects of prevention and treatment of human diseases, prevention and treatment of animal diseases, elimination of pathogenic bacteria in food, prevention and treatment of bacterial infection of crops, cracking of bacterial biofilm and the like.

Klebsiella pneumoniae is a gram-negative bacterium, widely exists in water and soil, and can cause various histopathological changes in intestinal tracts and respiratory tracts of human beings and animals. Antibiotics such as beta-lactam and quinolone are common clinical antibacterial drugs. In the face of the extensive use of these antibacterial drugs, klebsiella pneumoniae develops multiple resistance mechanisms.

Disclosure of Invention

The invention aims to provide a drug-resistant Klebsiella pneumoniae RDP-KP-20004 strain, and aims to solve the problems that poultry in a farm are not treated timely due to Klebsiella pneumoniae infection, the poultry is in an acute disease, and the drug use is ineffective due to drug resistance of pathogenic bacteria.

In order to achieve the purpose, the invention adopts the following technical scheme:

a drug-resistant Klebsiella pneumoniae RDP-KP-20004, which has a polyhedral three-dimensional symmetrical head, is wrapped by nucleic acid, has a diameter of about 70nm, has a tail with a length of about 170nm, has a tail sheath, and has a neck connected with the head and the tail, and is a tenoviridae, the phage is preserved in China general microbiological culture Collection center (CGMCC) 12, 17 days 2020, with the preservation addresses as follows: no. 3 of Xilu No. 1 of Beijing, Chaoyang, and the preservation number is CGMCC No. 21407.

Further, the phage can maintain good activity at pH 5-9, and its titer is 1 × 10⁶Above pfu/mL, at pH 7, the titer is highest, suitable for growth in neutral environment; at pH 12 and pH 3, the phage still survived, which was acid-base tolerant over a wide range.

Further, the phage has stable titer at 35-55 deg.C, and is maintained at 10 ℃¹⁰More than pfu/mL, still has biological activity after being acted for 30min at the high temperature of 80 ℃, and has better tolerance to temperature.

Furthermore, the bacteriophage RDP-KP-20004 can efficiently infect Klebsiella pneumoniae, and the optimal multiplicity of infection is 0.1.

The invention also provides the application of the Klebsiella pneumoniae RDP-KP-20004 in the preparation of medicines for preventing and/or treating diseases caused by Klebsiella pneumoniae.

The invention also provides application of the Klebsiella pneumoniae bacteriophage RDP-KP-20004 in preparing a feed for preventing and/or treating diseases caused by Klebsiella pneumoniae.

The invention also provides a sterilization composition for preventing and treating klebsiella pneumoniae, which comprises an effective amount of klebsiella pneumoniae RDP-KP-20004.

Further, the antibiotic for resisting Klebsiella pneumoniae is also included.

The invention also provides an additive for preventing and/or treating diseases caused by Klebsiella pneumoniae, and the effective component of the additive comprises Klebsiella pneumoniae RDP-KP-20004.

The invention also provides a cleaning agent or a disinfectant, which comprises the Klebsiella pneumoniae RDP-KP-20004.

The invention has the beneficial effects that:

(1) the Klebsiella pneumoniae BK-20004 is separated from the liver of a sick chicken in a breeding farm in Qingdao, namely Moita, and pathogenicity and drug resistance of the BK-20004 are determined through experiments, the BK-20004 is taken as a host, a Klebsiella pneumoniae phage is separated from the environment, the phage RDP-KP-20004 has strong cracking effect on the pathogenic Klebsiella pneumoniae BK-20004 in a poultry breeding environment, and a phage source is provided for the industrial production of phage for preventing and treating the pathogenic Klebsiella pneumoniae BK-20004 in the poultry breeding environment;

(2) the Klebsiella pneumoniae bacteriophage RDP-KP-20004 provided by the invention has good advantages in treating diseases infected with Klebsiella pneumoniae, and has high titer and good prevention and treatment effects.

(3) The Klebsiella pneumoniae RDP-KP-20004 provided by the invention has good application prospect in prevention and control of pathogenic proteobacteria of poultry.

Drawings

The above and other features, properties and advantages of the present invention will become more apparent from the following description of the embodiments with reference to the accompanying drawings:

FIG. 1 is an electron micrograph of bacteriophage RDP-KP-20004 according to the present invention;

FIG. 2 is a schematic representation of a one-step production curve for bacteriophage RDP-KP-20004 of the present invention;

FIG. 3 is a schematic diagram of the acid-base stability experiment of bacteriophage RDP-KP-20004 according to the present invention;

FIG. 4 is a schematic diagram of the thermal stability experiment of bacteriophage RDP-KP-20004 according to the present invention;

FIG. 5 is a schematic diagram of the genetic stability experiment of bacteriophage RDP-KP-20004 according to the present invention;

FIG. 6 is a schematic representation of the lysis curve of bacteriophage RDP-KP-20004 of the present invention against host bacterium BK-20004;

Detailed Description

The invention is further illustrated by the following figures and examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.

Example 1 isolation and preservation of Klebsiella pneumoniae BK-20004

Sampling from a horizontal breeding farm of the Qingdao, namely the inkland, taking the liver of a sick chicken in an aseptic operation, marking on a selective culture medium, culturing for 18-24h at 37 ℃, and forming thick, grey-white, round-convex and smooth bacterial colonies with wet surfaces on an LB (Luria Brookadside) culture medium by the pathogenic bacteria; forming thick and abundant pink viscous colony on the Macconkey agar culture medium, and having diffusivity; a white viscous precipitate formed in LB broth, which became uniformly cloudy after shaking; and selecting typical colonies, continuously streaking and purifying for 3-5 times until the colony morphology is consistent, then selecting a single colony, inoculating the single colony into 5mL of LB broth culture medium, and carrying out shaking culture at 37 ℃ and 200rpm for 12h to obtain a uniform and turbid bacterial suspension. And determining the pathogenic klebsiella pneumoniae by 16sRNA molecular identification and serotype identification, naming one strain as BK-20004, streaking the identified strain on an LB culture medium plate, culturing for 12h, scraping all colonies into 30% glycerol-LB liquid by using an inoculating ring, and storing in a refrigerator at-80 ℃.

Example 2 host bacteria BK-20004 drug susceptibility testing

The sensitivity test of six clinical commonly used antibiotics of ampicillin, ceftazidime, cefoperazone, kanamycin, cefepime and ciprofloxacin is carried out by performing standard M100-S17CLSI according to an antibiotic sensitivity test, and Escherichia coli ATCC 25922 is taken as a quality control strain for a resistance spectrum test. The specific method comprises the following steps:

(1) preparation of MHA medium: melting the sterilized MHA culture medium, cooling to about 50 ℃, pouring into a flat plate with the thickness of about 4mm, and standing for half an hour to solidify the culture medium.

(2) Preparing a bacterial liquid: 10. mu.L of the host BK-20004 and the quality-controlled strain were pipetted separately into 5mL of liquid LB and cultured at 37 ℃ for 12 hours with shaking at 200 rpm.

(3) Taking 6-6.5mL of liquid agar powder which is heated and melted, adding the liquid agar powder into 10mL of sterilized EP tube, sucking 200 mu L of bacterial liquid to be detected, adding the bacterial liquid into the EP tube, reversing the liquid agar powder, fully shaking the liquid agar powder evenly, directly pouring the liquid agar powder onto the surface of a culture medium on an MHA plate, covering a cover on the culture medium, horizontally placing the liquid agar powder in an incubator until the bacterial liquid is completely absorbed by the culture medium.

(4) The position for placing the antibiotic paper sheet is marked on the back of the MHA flat plate, and the antibiotic name is marked. The ultraclean workbench is subjected to ultraviolet disinfection, the tip part is shot by using outer flame of an alcohol lamp, the sterilized small paper sheets (circular filter paper) are clamped by using a buffer clamp and tiled on the surface of an MHA culture medium, 1 mu L of corresponding antibiotics are added on each small paper sheet, standing is carried out, and the antibiotics can be completely immersed into the paper sheets (in order to accurately observe results, the distance between the antibiotics in the paper sheets is required to be more than or equal to 24mm, and the paper sheets can be slightly pressed by using the clamp to ensure that the antibiotics are tightly attached to the culture medium). Parallel experiments were set up for each group.

(5) The MHA plate was placed upside down in an incubator at 37 ℃ and cultured for about 12 hours.

(6) After the culture is finished, the diameter of the inhibition zone is measured by a vernier caliper, the sensitivity of bacteria to antibiotics is judged according to the diameter, and the judgment standard is executed according to the diameter of the inhibition zone of the enterobacteriaceae bacteria (the judgment standard is shown in table 1, and the drug resistance experiment result is shown in table 2).

TABLE 1 judgment Standard of the diameter of the zone of inhibition of Enterobacteriaceae bacteria

TABLE 2 host bacteria resistance test results

As shown in Table 2, Klebsiella pneumoniae BK-20004 has strong drug resistance to clinically used cephalosporins, and has low sensitivity to kanamycin and ciprofloxacin drugs.

Example 3 isolation and characterization of phage RDP-KP-20004

(1) And (3) treatment of pathological materials: taking sick chickens from a farm, dissecting to obtain livers, shearing 2g of liver tissues under aseptic operation, adding 2mL of sterilized LB liquid, fully grinding the livers, centrifuging at 10000rpm for 30min, taking supernate, filtering the supernate with a 0.22-micron filter, and keeping the filtrate for later use;

(2) preparing phage enrichment solution: adding 0.2mL of the bacterial suspension and 1mL of the filtrate into 5mL of LB broth, performing shaking culture at 37 ℃ and 100rpm overnight, then centrifuging at 10000rpm for 5min, taking the supernatant, and filtering the supernatant through a 0.22 mu m filter to obtain a filtrate for later use;

(3) phage separation: separating phage by double-layer plate method, mixing 0.1mL of phage enrichment filtrate with 0.2mL of host Klebsiella bacterial suspension uniformly, water bathing at 37 ℃ for 10min, spreading double plates, culturing in 37 ℃ incubator for 6-8h, soaking the transparent spots in 1mL of SM buffer solution at 4 ℃ for 12h, taking 0.1mL of leachate and 0.2mL of bacterial suspension, water bathing at 37 ℃ for 10min, spreading double plates, culturing in 37 ℃ incubator for 4-6h, purifying, and purifying 5 times according to the steps to obtain the product. The plaque diameter formed by RDP-KP-20004 on the double-plate is 3mm-5 mm.

The method for preserving the bacteriophage RDP-KP-20004 comprises the following steps: phage multiplication fluid was mixed with 60% glycerol according to 1:1, and storing in a refrigerator at-80 ℃ or liquid nitrogen.

EXAMPLE 4 Electron microscopy of phages

Dropping 20 μ L of liquid containing crude phage particles on a copper net, settling naturally for 15min, sucking off the excess liquid from the side with filter paper, adding a drop of 2% phosphotungstic acid (PTA) on the copper net to stain the phage for 10min, sucking off the staining solution from the side with filter paper, drying the sample, and observing the form of the phage with an electron microscope, as shown in FIG. 1.

The bacteriophage RDP-KP-20004 has a polyhedral head part with three-dimensional symmetry, which is wrapped by nucleic acid, has a diameter of about 70nm, a tail part with a length of about 170nm, and a tail sheath, wherein the neck part is connected with the head part and the tail part. According to the ninth report of the organization of viral classifications by International taxonomy of viruses, this phage was classified in the Myoviridae family of the order Naphilae.

Example 5 phage optimal multiplicity of infection assay

The phage multiplication solution and the host were added to the LB broth at a ratio of the multiplicity of infection of 100, 10, 1, 0.1, 0.01, and the total volume of the culture system was ensured to be the same. After shaking culture at 37 ℃ and 200rpm for 8h, centrifuging at 12000r/min at normal temperature for 5min, and spreading the supernatant on a double-plate to determine the titer. See table 3 for details.

TABLE 3 determination of optimal multiplicity of infection

The data in Table 3 show that when the multiplicity of infection is 0.1, the proliferation fluid is relatively clear and the titer is highest after 8h of culture, which indicates that the optimum multiplicity of infection of RDP-KP-20004 is 0.1.

EXAMPLE 6 one step growth Curve determination

Taking Klebsiella pneumoniae suspension in logarithmic growth phase, inoculating Klebsiella pneumoniae BK-20004 and bacteriophage RDP-KP-20004 according to the optimal complex number of infection, carrying out water bath for 10min at 37 ℃, centrifuging for 5min at 12000r/min at normal temperature, removing supernatant, removing free bacteriophage which is not adsorbed on a host, and washing twice by using LB broth culture medium. The pellet was resuspended in LB broth at 37 ℃ and quickly placed on a shaker at 37 ℃ and 200r/min for shaking culture and timing. Sampling and counting every 5min for the first 30min, counting every 20min for 30-60min, counting every 30min for 60-300min, and drawing a growth curve by taking time as a horizontal coordinate and taking a bacteriophage titer logarithmic value as a vertical coordinate, as shown in figure 2. And obtaining the incubation period and the lysis period of the phage, and calculating the average lysis amount. Mean lysis is end phage titer/initial host bacteria concentration.

As can be seen from FIG. 2, the titer does not change significantly within 60min after the phage infects the host bacteria, indicating that the incubation period is about 60min, and the titer of the phage increases significantly within 60-120min after infection, then becomes stable, indicating that the phage lysis period is about 60 min. By calculation, the phage lysis amount is about 160 PFU/infected cell, which shows that the phage has extremely strong lysis and replication capacity.

Example 7pH stability assay

Preparation 10⁵cfu/mL of host bacteria liquid in exponential phase, and adjusting the concentration of the phage liquid according to the optimal complex number of infection. Adjusting the pH values of LB liquid culture medium to be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12 respectively, adding phage liquid into the LB liquid culture medium with the same volume and different pH values, uniformly mixing, standing in an incubator at 37 ℃ for 2 hours, mixing 100 mu L of mixed liquid with the same volume of bacterial liquid, measuring the titer by a double-layer plate method, drawing a pH-titer curve and determining the pH range of the highest point, wherein three pH values are parallel. As shown in fig. 3.

As shown in FIG. 3, the phage RDP-KP-20004 can maintain a good activity at pH5.0-9.0, and maintain a high titer, and the phage titer is highest at pH 7.0. The titer of the phage is obviously reduced along with the increase or decrease of the pH value; the result shows that although the bacteriophage RDP-KP-20004 can not tolerate strong acid and strong base, the bacteriophage RDP-KP-20004 can tolerate weak acid and weak base well.

EXAMPLE 8 optimum growth temperature determination

Preparation 10⁵Diluting phage stock solution to a proper concentration according to the optimal infection complex number of cfu/mL host bacterial solution, mixing 100 mu L of phage stock solution with the bacterial solution with the same volume, pouring a double-layer flat plate, culturing at 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃ and 80 ℃ for 30min and 60min respectively, measuring phage titer by a double-layer plate method, drawing a temperature-titer curve for three parallel temperatures, and determining the temperature range of the highest point. As shown in fig. 4.

As can be seen from FIG. 4, the phage was stable in titer at a temperature range of 35-55 ℃ and maintained at 10¹⁰The pfu/mL is above, still has biological activity after being acted for 30min at the high temperature of 80 ℃, and shows that the bacteriophage can tolerate higher temperature in a short time.

Example 9 genetic stability test

Continuously passaging RDP-KP-20004 for 29 times, adding equal amounts of RDP-KP-20004 and KP-20004 into each generation of culture medium, culturing for 6h for each generation, pouring the double plates, and measuring the titer. As shown in fig. 5.

As can be seen from FIG. 5, the phage RDP-KP-20004 always maintains higher titer in the process of 29 passages without obvious fluctuation, which indicates that the phage can be stably inherited

Example 10 phage vs host lysis curves

The lysis curve of the phage on the host is obtained by adding the phage to the bacterial suspension, and the OD600 value is changed due to the lysis effect of the phage on the host. The host bacterial suspension cultured for 6 hours was inoculated into 100mL of LB medium at a ratio of 1:100, to which phage-proliferating solution was added at the optimum multiplicity of infection, while 3 replicates of each group were set with no phage added as a control, and the results were averaged. The results are shown in FIG. 6.

As can be seen from FIG. 6, the OD600 of the host without adding phage gradually increases within 40min and exponentially increases within 60-120min and becomes stable after 240min, while the host with adding phage gradually increases OD600 at the stage of adsorption and infection of the host within the first 40min, and the increase of phage also shows exponential form with the exponential increase of host bacteria, and the OD600 does not obviously increase within 100min but reaches the peak with the time increase, and the host bacteria without cracking continue to propagate and the OD600 gradually increases.

It should be noted that the specific embodiments are merely representative examples of the present invention, and it is obvious that the technical solution of the present invention is not limited to the above-mentioned examples, and many variations are possible. Those skilled in the art, having the benefit of this disclosure and the benefit of this written description, will appreciate that other embodiments can be devised which do not depart from the specific details disclosed herein.

Sequence listing

<110> Ray union (Qingdao) bioengineering Co., Ltd

<120> drug-resistant Klebsiella pneumoniae RDP-KP-20004 and application thereof

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<170> SIPOSequenceListing 1.0

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atcatcggaa cttttaacag tatgttgatg atgcagcgcc taaaagatgc taaagtatcg 4980

cattactttt ggttcgggat gatggacgac cgcgagcgcg agagccatat caagctagaa 5040

ggtaagcgac gccccgttaa tggtgacggc attttccccg gtgaagagta cggttgccgt 5100

tgttgggcgg ttccagattt taacaatgta gaggtatcat gatgaaaaga gttcaaaggt 5160

tcgacacggt aaagatgaag gcccgattcg atgagaacgg ctttctggtt gatactccga 5220

tcgtggcgcg tatcggtgcg cagacgtacc agacgccaac cgggccgcgc gtcgagttcc 5280

gcccgcgttc ggaagtgttt gatgctgaat cactggcttc ataccagggc aagccgatca 5340

ctttgggtca caagatggtg aacgcgcaga acgcaaaagg cctggtcgtt ggctcgtgtt 5400

ccggcgctgg caaagaagag gggatcggcg ttcttgttcc ggtgatgatt tacgacggcg 5460

agtcaatcga gcaagccaaa aagcgcgtag cggctgagtt atccgtgggc tacacttcgg 5520

ttgatatcga tcgcaaaggt tggggtaata acgcaactgg cgaatattat ttcgatgaag 5580

accttccgga aaacttcgaa gagatgaaaa atgattccgt ctctgattgg gttcgctttg 5640

atgcggtgca aacgaagatt cgcgtgaatc acgtcgcgct tgttttccgt ggtcgtgccg 5700

ggattgcgaa attaaatctt gatagcgaac aagaattccc ctatgatgac gactcaaacc 5760

acaaaggagc taaaacaatg atcattaaaa ttgacggcgt agatgttgaa gtggccgata 5820

acgtaggcgc ttacattgcc aaactagacg cgcaagttgc atcagcaacc agtcaggtaa 5880

ctagcatcac cgcagagcgt gacgcgcttc aggccaaagt tgatggcatt gaagatgaag 5940

ttgccgcccg cgttgctaaa atcaaagccg acgaagacgc aaaacagaaa gttatcgcgg 6000

tggtttctgc cgctggcgtc aagtgcgacg gcctggatgt taaggcgatg aaggttgctt 6060

acatcaaaga ggtagacggt cgtgatctgt ctgacaaaga agattcgtac atcgacgctt 6120

cttttgactt tatcgccaac tctgataaga tggctggcaa tcgctccaaa gtcttcggca 6180

aaaaagaaga tggcgagcaa aaagacaaag gcggcttacc gaaacttgac ggcaccgaaa 6240

tcatcgatcc gcaggcaaaa ttccgccgct aataatttgc ggccttcggg ccgctaccag 6300

aacaaataaa caggagattc aaaatggcac agattccagc ttcttattcc cgcaagcgcg 6360

atattgccgt agcggggcag atcgctgata cgtcgcttta taatatcgac ggcacttgtg 6420

ttgctgaagg tgatatcaaa gctggcgtac tggtggcttc cactggcgca gtttccaatg 6480

gccacaaggt cgttaaacca gcaaccgcag cgagcgacgt cattgtaggc atcgcgcagt 6540

tctcgcaagc ctactcgcct gaaggcaagt atgacgatga gagcgcagtt aacgttatga 6600

ctcatggccg catttgggcg atcgctgacg caactgttac agaagcagat tgcgcgttcg 6660

gttctttcgt tacctttacc gcaactggca ccgtagcgaa gggtgatgcc ggggttatca 6720

aaactagcta caaacacact ggcgaataca ccaaaaacgc agatggcact gttctggtga 6780

aagttcaggt gcttcagggc gcggtggctc cggcggctgc ggctggcggc ggcgctggtg 6840

cataataaat ggtggggctt cggccccatt tttgcaccag aaaaaaaact ttgacggctt 6900

aacgattcgt gatattcttc atctcgttaa gctaaataca caaacaggag ttttcagatg 6960

actatgaaat tagatgcatt cgaacagagt gccattaagg tcgcaatgca gggcatgggc 7020

gtcgacgccg caaaactgga tgcttacggt atctggactg ttaagcagat gactcaatta 7080

ctgaatcgcc agtatgagca ggcatacccg cagaccagcg cacttgagct tttcccggta 7140

actaccgaga tctcgccaac cgcccgccgc tttgagtggc tcgaattcga tggcgtaact 7200

tctgcgaaaa ttatcgccga ttacaccgac gatctgccga ccgttgaagc gatggcgaaa 7260

gagaagtcag ggaaagtttt ccgcctgggt aacgcgtggt ttatttccat cgacgaaatt 7320

aaaactggcg cggcgctggg ttccagcctg agcgatcgca aggcaactct ggcccgcgaa 7380

ggtcatgaga cgctcgttaa tgatctggtg ttcaaaggct ccgctcctca cggcatcgtg 7440

agcgttttcg accatccgaa cattaaccgc atgaccgcca gcgcggcttg gggcgatgac 7500

gcagcagcgg ctgaaaaggc attcgaagat ctggaagacc tgctaaacat gatcgaagaa 7560

actacgctgg gccgccatca cgcgaccaac atcgtaattc ctccgtctaa gcgtcgtctt 7620

ctgacgaaaa agatgccgga cactagcggc gactcttatc tgacctggtt caccaagaac 7680

cacccgaaca tcaccattac ggcgatggcg gagctggaag atattgacgg cgcaggcacc 7740

aaaggcgtgc tggcatacga aaaagaccca atgaacatga gcatcgagat ccctgagcgg 7800

ttcaacatgc tgccgatgca gccgaaagac ctgcatttca aagttccgtg cacctccaaa 7860

tgcactggtc tcatcgttta ccgcccgctg actattgcga ttctcaccgg gatttaatca 7920

aaaagcgcct tcgggcgctt ttttattgca ttgcatccta caatgtgctt taatttgaaa 7980

cctaaagtaa accaatggag cattaacaat ggccagtaaa aaagaaaccg tagaaaccgt 8040

cgaaaccgta gaaaccacca gtgccgaaca ggcgtcgcag gctgttcaac tacaaaacgt 8100

tggtgcatgt gcaattcgct ataaaggcaa aaattacgtc tatgagcagg tttttgaagt 8160

gccggaaaac gaaatcgacc gcttccgcca cgaaatcttc aaaggccgtg tcgaattcta 8220

cgacaacccg aaacgcacgc gcgaatacat cgcagcagtg aaggcgaaag cgaaagagat 8280

cgtgcagcct aaaagcgcgg aataacaaaa accaacaaag ggcgcttcgg cgtccttttt 8340

catatcagga gatcgacaat gaattacacg attcaagatg tgatcgacaa gatgcgcagc 8400

cttgcgcctc cgcttaaaaa agtacccgac gaactgctaa ccgcatgggt cgtgcttgcc 8460

gaggagttcg tttgcaaatc aaggttcggg gattccattg ttacagcgat cgcacttatg 8520

accatgcatc taatgttttt ggatggcgca atgaagcaag aaggcgaaag cctggagtct 8580

tacacgcagc gagtggcatc gttcaccctg accggggagt tttcccaaac attcgatcgc 8640

gtatcggcgt caagcgacaa cgaaatgctt tctacgccgt ggggcaaaat gtactggcgt 8700

atgctcaaga tgcgcggcgg cggcttcggc cttcttaccg ccgggaacgt tcgccgctgc 8760

ggaattggta ggtgatcgct atgaactaca aacaaatcca ggcacgcgca agcgcgggca 8820

ttaagttttt cagtgacgct gacggcgtat tcaacaagta cacgaaaggc gctggcggcg 8880

gaatcgatcc ggaaaccggg gaagatatca ttcctggtga ggtggtaacg acaatcaagg 8940

gcgcgatcag ggatgtaaat gaccgtgaca ttaacggcga aaccatcctc gctggcgaca 9000

agcgcgggtt cttcactcac gatgtgccaa tcatggaggg tgacgaaatc gaagtagacg 9060

gcgagcgcta ccgcgtggtt aatgcccgtc cggtaaaacc aacgggaacc gttgtcgcct 9120

accgtccagt tttacgcagg gtggcgactt atggctaatt atacgatccg ggagttcaca 9180

ggggctattg atgcatggtg taaggccgct ggtgatgcgc tggaggacgt tgtaaggttt 9240

acgtgtgaag atattcaccg cgaccttgta atgcgttctc cggtggatac aggtcgcttc 9300

cgtggtaact ggcaaatcac ctttaaccgt gcagcgcttt acgcgattaa cgcatatgac 9360

caaactggcg aaaagacgat ccagaacggc aaagccaaca ttgcactata cgcaaaaggg 9420

gctgggatca cttcgatctg gttcagtaac atgctaatct atgcgaacgc gctggaatac 9480

ggccattcaa agcaggctcc caatggcgtt atgggcgttg ttgcgatccg gttgggcgtt 9540

tatgttactg aagcaatcaa gcgagcgagg gcgaaaaatg cattatgata tggcgttaaa 9600

atgcaaggcg gcagtggcta aatttgccgc cgagaacggg ttgaggttgg cgggggataa 9660

cgttgacttt atcccaccga gaggcgggga aacctacctt aaagcctcct acgtcgaggc 9720

ggattcaaga tcggttgacc tgtcaaggaa atgccgagtc tatctggcga tggttcagat 9780

tgacgttatc tttaagcctg ggatcggaac cgaccgcgcg aggattatcg cccaacgcgt 9840

tgcaaaatcc ttccctgaag gaaagattgt tgatcgtgac ggtaagttat atgtgagcga 9900

gtgggcggag gtacacggcg tgcagaagca tgaggctggt tggttctttc cggttcggtt 9960

cacggtaaga tgcgagagca ttgaggaaaa cggttatcca tcaacctgac gggtcttaga 10020

ggtgcttata atttcttgcc agcctggaaa tatataggca taatggcgtt gttaaacttt 10080

catcaaaaca ggagtattca acatgcattt accaaacggt gcaaaggtct tctttgagaa 10140

ggcccgcggt gcggaggttc cgtttaccgc aatgaccaac gacgcgaaaa acccaacaat 10200

cacggtggca gatggtgcgc tcgacgttaa cgatatcgta atctttaccg attgcacttg 10260

gagcgacttt gtaaataaag tggcccgcgt caagtcggta tcggctggtg tggcaacgct 10320

ggaagagttc gacacctccg acactaacaa gtatcctggc ggcgcggcca ctggtagcgt 10380

gagcgtgatc tctgattggg tcgaattgcc ttgcattcag gatttaggca aagacggcaa 10440

cgaacagcag ttctataact atcagtgcct tggcgatgag cgcgagcaat ccgaacctac 10500

ttacaagtcg gcggtgacgc ttaactacac gtttgcgcac gattacagta acgcgatcta 10560

ccctgtgttg cgttcagccg acgccagcaa gcaggcgaaa gcgatgtaca tgtatatccc 10620

gcgagcttcc gaggttcgtt actggtctgg aattgcatct ttcgatgaca ttccttccac 10680

ggctgttaac gagatggaaa cggtaacgct taacattgcg cttaaaggtg cgcacgtctt 10740

ccttccggtt gtcgcgtaat taaatggcgg ggcttgtgcc tcgccttttt ttgtgcataa 10800

tagcgaataa cacaaaccaa tcaggagtta acaaaatggc taaattcaaa attcaaattg 10860

gcggcaacct cccttctttc aagctgcccg taaccttcac ttgcccggat ggcaaagaag 10920

caaccatcac catgaccgta aaacaccgct ccaccgatga gatgaaagac ttctatgaga 10980

gcgaagataa agcgccaaag ggtaacgccg agtttatccg cttcatggct gaaggttggg 11040

atctggatga cgaattcagc gatgaaaaca tttcctggct ttgcgctcac ttcccggcat 11100

ttgtcatggc actgccacaa acttacatgg ccgcacttgc gggccaccgt gcaaaagttt 11160

aaggcgggct gtttatctca cgcttcagcc tgagctaacc gatcgccagc ttgcggagta 11220

cgggttaagg cgatcggact atgaagcaga tcttgaaacg atatattttg atgagcagac 11280

cgcccaaagc tggcagctat tccaggccat gcaaacgcaa tggcgaatcg ggatgaatgg 11340

cccgacgggg cttgactata atacgttgcc actgctgttc gaattgtata aaatcgacaa 11400

tcgagaagcg gcattacttg acttgcaaat cttagagggt gaatacctga aggagattta 11460

caagaaatcc aaataagcgc ctacgggcgc ttttttcata tgggggctaa acatggctga 11520

taaagtagct gggttgacgt ttggcgttga cgtttcgcag gttgacaaag cggtacgatc 11580

actcgcagaa ctgaaaaacc aaagccagca aacgggcgct ggcctacagt cacttgcgga 11640

cgctgaaagg cgggccacgg cgcagaccga ggaaatgaac cgcgcgttgc agcgccagaa 11700

gcaagagacg gacaaatcaa aaaccagctt tagcaggatc gcaagcgcca tcgatcccac 11760

gatctcaaaa atggccaact tgcgcaaagc gacggaagaa cttgataaag cgtgggcttt 11820

ggggcttgtt ccagataagg aatttttccg ccttggtgct atcatcgaat ctactaccaa 11880

ccaactccgg cggcagcagg cggcgctaac cgaagaaggt cgcgcagcaa tcgcagaggc 11940

ggaggcaaag cagaaggcgg caaacgccgg gcgtgatttt gtcgccagtc tgaaacagca 12000

ggctgaatcc atcgggaaaa cacgcgccga actgctggaa atgaaggcgg cgcaattggg 12060

cgtgtcggca gaagcggcac cgttcattaa tgccctgaag cagcaggagc aagcattaaa 12120

gaaacagcaa aacgctatgg gcctcgctgg catttctgcc gggcaatata aaatggccat 12180

gcgccagtta ccgatgcaga tcacggatat tgtaacgtcc cttgcgtccg ggatgccaat 12240

ctggttggtt gctgtgcagc agggcgggca aatcaaggat agcttcggcg gcttgagcaa 12300

tacgtttaag gtattgttga gctacatcaa cccgctaacg gttggtgctg cgtcgttggg 12360

tatcgccctg gcggcaatcg ctaaagctgg ttatgactcc tggagtgcgc aacgagaact 12420

ggcgaatgcg ctggtgctga ctggtggcta tgctgcgacc accaccgggc agatcacagc 12480

cttaaccgaa gagattaaca aaaactcatc ggctacgatt ggcagcattc aggaaatcgc 12540

aacatcgcta gctagttctg gtaagtacac aatcaaccag attaagcaga tcacgaagac 12600

tacggaggaa tggtcggcgc agacgggtga gagcgagaag acgatcacag ggtattttga 12660

ttcgatcgta aaagaccctg ttaaggggct tgccgatctc aatgagagct ttaattttct 12720

gaaagagggc cagctaacct acatcgaatc tttgcgtaaa accaaaggcg agacggcagc 12780

agtagatgcg gcgacaaaac tgtttgctga aacgatggat aagcgcctta aagatatcgc 12840

ggacagcgca acgccgctag aaacgatgtg gacggatatc aaaaaatggg ccgctgactc 12900

atggaaatgg gtaggcgatc acacggtcgg ggcgctaaac cttatcgttg atacggtttc 12960

cgcgatcatt aacacgatca ggaagctgat tgccgacggc gacgccatga tcgcgcagtt 13020

catcgttgac gcaggccgga aactgcaaaa aattcccggg atgggcgact ttggcaatga 13080

atttctggcg cagcaggagc agctaatcaa ggactcgaaa gccaaatccg acacgcttgc 13140

aaaaaccatt gcggaacagc aggcaaggat cgcaaaaggc gagatgggtt acattgatgc 13200

cgcgaaggat aaaacccttt ctggtgggca cagcagcaaa acgaaggatc gcgttaacca 13260

ggaagaaaaa gatatcctga aaaaccgcaa cgccaggaag cagcaggtag acgcgggcgt 13320

taaaattgat gagcagtacc aggctgaact gctatcgcta caggcgcagc taaaggtttt 13380

gcagcagcat aaagggcttg acgacaagat cagccagcag cgcaaagact acttcgagac 13440

ggtcgccaaa tttcaggtgc tggaagaggc aagccagaag cgaaaactga accaaagcga 13500

acagcagatg ctggcgaaca aaaagaatat cctgtacatg gcggaacaaa aggccatcgt 13560

gggagatcag attgttcagc agcagcgact gaacgccttg cttgacaagt ctaccaagta 13620

tcaaaaccag atggcggaga aaaccaaagc gctacaggat accgccggaa tgggtagcaa 13680

ggagcaggag agataccggg ccaatgcgca gatggcggct gactggaaaa acagcggcgg 13740

atctttaagt gaccctggat ttaaagccat gcaggatgca agtaataaat tctacgcaca 13800

gcaagatgcg caaatgatga actggaaggc cgggtttact cacgcgtggg ctgacattgg 13860

cgacgaagtt aatgacgtat acaccaacat cggggatatt actaaaaacg catttagcgg 13920

aatggcgagc gtgctgactg attttgtcat gactggtaag gccagcttta gcgactttgc 13980

aaagagcgtg atcactgaca ttaccaacat gcttatcaag atggcgctgt ttaactcgct 14040

ttctgctgcg tttggtggtg gtggcggcac gttcagcttc gccaacatgt tcagcaaggg 14100

attttccggc ggtggttaca ctggcgacgg agggaagtat gagccaaaag gcgttgttca 14160

cggcggggaa ttcgtattca ccaaagaggc gacgcaaagg ttaggcccgg aaaacctgta 14220

ccgactcatg cgcggctatg caagtggcgg cctggttggc ggtaacgcaa gttcagtatc 14280

tggagtaacc aacggcggca atgttgcggc gtcggctgcg atggtgttca ctatcggtga 14340

cattaacatt acgatgggct ccggtcagga tagtaagggc atggagcaag gcgtgaggca 14400

gatcgtgaac gatatgttca ccgaggcttt gagccaaaac gggcgcattg cgaagttcgt 14460

caatgagaaa acgaggaggt agtagtggat tcctttaaat ggtgtacaca aattcaagga 14520

ggggcggcaa aggtcgccat ctccaacaac attcgatcga ttgttttcgg aaacggctac 14580

atccaaacgg catcgagcgg cattaacaca aagcgcagga cggttccgat cgtttatggc 14640

ggttcagatt gggaggaggt ttataacttc tgtcaggatc acgtaacaaa gccttttgtg 14700

tggaaggcac cggatggaag aatgggcgta ttcgtcgtaa ctgccgactc cgttaacctt 14760

gcgccgcagg gtggcggggt gtttgaggta acggctgaat tcgccgaacg cttcacttca 14820

gccggataat caaaaagcgc cctttacggg tgcttttttt tggcctatga tctggagtca 14880

attagaggag ggcttacgat gacagccaat gtttcaaaag agtttgcgaa ctgcttacaa 14940

aaacttttcc ccggcgagat cctaacgctg atcgatatcg acgccacaaa gttcggcggg 15000

caggtctacc gattccataa cgagaacgtc gcctatacaa ccgaggagct tttggcagcg 15060

gttaacggcg ggacgcttaa gccaaagatg atcacgtttc gcggcgagca gtacggcccg 15120

cgcccgttcg gcctgggcgg gatcgcaatg tcgagcgatg gcacagtgga aaagccaacg 15180

ctgacggtta gcaatatcga tgcacaagcg agtgctctta ttcgcgccta caacggcctc 15240

atgcaagcca aagttacggt atgggttttg gtcaaggaat tgctacaaaa cgacggcagc 15300

gttaaagagg gcgattttag gcgatttgtc tactacatcg agcgcccaaa acaggtcgac 15360

ccgcaaaagg caacgttcga attaacatcc gtgtttgata tggatggatt aatgatcccg 15420

gcacgcctaa cgcaaaccgt ttgttattgg gcgcagcgtg ggtggtacaa gtctggcaaa 15480

ggctgcgact acaacgggca gaacggatac ttcgacaagt tagggaacag ggttgacgat 15540

ccgtcgcggg atgtttgcgg tggcctggtg tcttcttgca ggcttcgttt tggtaatgaa 15600

ccgttgagtt tcggcggttg tgcgacagca actttgaaga gtggtagcta atatgttgac 15660

tccgaaaatt aaaatgcaga tcatgcagca cgcgaaggaa gtctacccgc acgaatgcgc 15720

cgggctggtg acgcaaaaat cacgcgtgca aaaatatcac cgactcgaca acgtttcgcc 15780

agatcctgag aacgagtcta tgccggacga aacgcagtat gcgatggcag caatggatgg 15840

ggaaccgatc gccttcgttc attcccacac tggcgacggt gcaaccacaa ttccgagcgc 15900

cactgattta tgcttctgtg atgagtctgg cttgtcgtgg gttatcgtct ccatcccgga 15960

aggtgatatg cgaatcattg agccgaagcg ccgcccgctg attggtcgcc cctgggcttt 16020

gggtgcttat gattgctatg gccttattat ggattttcac aagcgccacg gcgtcacgct 16080

aaaagatcgg cgtgttccgt tcgaatggtg gaagccggaa tacaaagaga atctttacca 16140

ggactactgg caagaggacg ggttcattga aaacactggc gagcctgaag ttggcgatat 16200

gatcattttt cagcttcagg cggagaagtg gaatcacgcg gggatttacg ttggaaataa 16260

caacatcctt catcacgcat atggcaagct gtctcgccgg gatatctatt ctggatggta 16320

cgagcagcac aaggttttaa tttgcaggca taaggagcta aaacatggca tcacatacaa 16380

agacgattaa actatctggt tccctgggcc gtcggttcgg tgtcttccac aaacttgcgg 16440

ttgattcagt cgctgagtgt atccgggcgc tgtcttacca ggttgaaggg tttaagccgt 16500

tcatgcagag caaagttggt tcaaacatgc gcttcggtat catcgcagac ggaaaaccaa 16560

tcagcacgga cgactttgct actttcgccg tggcaaggga gatcagaatc atcccgatcc 16620

caagagccag aaagaacggc gggttgttgc aggtcgttat cggagcggcg attatggttg 16680

cggccttctt tactggcggc ggttcgctgg cggctatggg cgctttttcg tcggcggctt 16740

ttatggctgg cggctcaatg gttttgggtg gcgtaatgca gatgatcgca ccgcagatgg 16800

gcggcaacat gcgtgcaagc gaatcacctg agaacaagcc atcttatgcg ttcggagggc 16860

cgattaacac cacggcggcg ggttatccaa tccagttgcc atacggttac agattggctg 16920

gcggcgcgtt gttcggttca ggatcttacg cagaagacaa caactaatta agccattcgc 16980

tttttagcct gggggcgtag cctccgggct ttttgtcgtg tacaattgcg agactattaa 17040

caggaggcta aacgatgact aatatcaagg cccgcaaggg tggttcaagc aagccacgta 17100

ctcccgtaga aatgccagac aacctgatct caaaagataa gatcaagttg ttgcttgccg 17160

tttcggatgg cgaggtggtt aacgacttca gcctgaagca gttgcatttt ggaggcgtcc 17220

cggttcagaa cgaggatgga acattcaact atgagggcgt gattgcagag ttccgccccg 17280

gcacgcaaac gcaggactac attcagggct tcagcgagtc aagtgctgaa ttccaggttg 17340

ctcgtgaagt cactcacaac acgccttata cgcttaccgt atcgaacaaa aatctttctg 17400

ctattcgctt tcgcctgtta tggccgcgcg tgctgactca aaaagataac ggcgatatgg 17460

tcggatcggt tgttgagtac aagatcgaga tggcggtaga tggtgcaagt tatcagacct 17520

acataactgg caagattgac ggtaagaaca cgactggcgg ttacgatcgg agcattcgcg 17580

ttaacctgcc gcaaaacttc acgtcgcagg tgcttatccg cgttagtcga gtaacgccgg 17640

acgctgacgg ggtgaaagtt gtcgacgctt tccgggttga atcctacgct gaagttattg 17700

atgcaaaatt ccgctacccg ttaacggcca tgctttacgt tgagttcgat agcgatctgt 17760

tccagaacca gatccccact atctcactca aaaagaaatg gaagattatc caggttccga 17820

gaaactacga tccgattaat cgcacgtact ccggaacgtg ggacggtgtt ttcaagtggg 17880

cgtggagcaa taacccggcc tgggttcttt atgatctgat catgaatcag cgctatggtt 17940

tagaccagcg tgagttagga atcccggttg ataagtggtc gctgtatgag gtggcgcaat 18000

attgtgatga acttgttcct gacaatcgcg gcgggatgga accgcgctat ttaatggatg 18060

taattgttca gtcgcaggtt gaggcgttcc agttggtaag ggatatttgt tccgcattcc 18120

gtggaatgac gttctacaac ggtgaaagcc tatcgatcat cgtcgataag ccgcgcgatc 18180

ctgtgtacct gtttacggct gataacgtcg ttgatggcgt tttcgttcgg acgttcccaa 18240

gcgaaaagac gatgtacacg tcgtgcaacg tcatgttcga cgacgaagaa aaccagtacg 18300

aacaggatgt tgaaccagtg ttcaacccgg acgcagcaat gcgatttggc cacaacccga 18360

ccagcattac agcgatcgga tgcaccagaa ggacggaggc gaaccgccgt gggcgttgga 18420

ttctgcaaac gaacctaagc gccacaaccg tttcgttttc tactggcctg gaaggtatga 18480

ttccttcttg cggcgatgtt atttacgttg cagatccgca ctggcaatcg gcctttaacc 18540

tggtgctatc aggacgcgtt atggaagtat ctggcgtgca ggtgttcctg gcctaccgct 18600

gcgacgcgaa ggctggcgat actctgatcc tgaataccga cgacggcaag cctgtgcgcc 18660

gcacaatcgc cagcgtttcg gctgacggta aaaccctaac gctaaacgtg ggatttaatt 18720

ttgacgttgc gccggacagt gtattcctga tcgagagcga tcagcttgca gcggaacagt 18780

atgtagtaac ccggattgaa aagggtagtg atgacgacga attcaccttt gccatcacgg 18840

ctacgcagta cgatccgaac aagtatgacg cgatcgacaa cggggtaatt accgatagcc 18900

gaccaacttc ggttgtagac ccggattcat tgggcgctcc gaaagactta acgattagct 18960

cgttttctcg cattgttcag ggaatgagcg tcgaaacgat ggtgatcggc tggtctgctg 19020

tacagtatgc caaactgtac gaggtacaat ggcgcaagga tggaggtaac tggaacaacg 19080

ttccgcgcac tgcgacaacg caggttgata tcgaaggcat ctatgctggc gagtaccagg 19140

cccgcgttag gtgcattagc ggcgggaatg tagcgtctcc gtggtcttct ttggctagtg 19200

catcgctgac cgggaaagtc ggagcgccaa aaggcccgat taaccttttt gcgtctgaca 19260

acgagatctt cggcattcgc gttaagtggg ccatgccaga aggggcggaa gacacggcat 19320

acattgagct ttaccagtcg caaagcggaa ccgatcagga cgcaagcctg cttaccctga 19380

ttccctaccc ggcggctgaa tactggcatt caattttgcc tgctggctac gttaacttct 19440

ataaggctcg aagcgtagac cgtatcggca acgtttcagc gtggactgat tacgctcgcg 19500

gcatgtcgtc tactgacgtt aacgccatca cggatgtaat cttggatgag atcctcgaca 19560

gcgacgcgat gaaagaactt caggtgagtg cgcaggatag tgcggcaaaa ctcaatgaat 19620

acgctaacag cataatccaa aacgcattag cgaatgatgg agatgttaga atcatgagaa 19680

aggagaatgg aaagaggaaa gccgaaatta aacatgcaga agttctcata gcaaatgaga 19740

cggaagccag ggtgcagcag gttaatcaga tctcggcaga gttcaacgaa aatctcaatg 19800

ctggattaac tcaagttaac gaggcgctgg ccaatgaaac tgaggctaga gttacttcgg 19860

aagaggcgct ttcagcaagg atcggagaga actccgcagc gctagatcag aaactcgact 19920

cgtgggcaga cgttaatggc gttggttcca tgtatacgat gaagctgggc ttgaagtaca 19980

acggccagga atataattcc gggatggctc tacagcttac cgcgcaaggg agcaacgttg 20040

tttcgcaggt gcttttcatt gctgatagat ttgctattat ccgtaatgca gcatccggcg 20100

cgtacacgct accgtttgtt gtgcagaatg accaggtgtt catgaataac gcgctgattc 20160

aggatggctc aatcaccaac gcgaagatcg gtaacgtcat tcagtcaaac aactatgttt 20220

caggtagtcg aggatggtca atagacaaaa acggaacctc tgagttaaat gaggtaacag 20280

tgagggggac aatttacgct actgatggtt ggtttaaggg tactgtttat gcagagcata 20340

ttgaaggtga cgtgatgatc gcagaatcaa acacgatacc attcaagaaa tattcagatg 20400

ttgatagcgg agattatgaa atactgaaaa ttcatggaga aaactttgat aggagtatcg 20460

atacgaacct gattatacaa gtgaattcta cattcaggaa tacttttaga gtaattgttc 20520

agactccagg gaagggggaa atagaattct acagtataga cactggaaat gacggaggta 20580

caagagcata cgcgctacgt ggcttcgatt tgccagcagc tggaagaggg caagagaata 20640

ggatcatcgt tagggtaact caaagcggga ggggtagcag cataaaaaca ttcactccaa 20700

gggttgagag agataatgga agcactggac aagaaatgtc tgatggagtc atatataaca 20760

ctggttttat cagagagagg gcatacatat cggcgtacag gaaaggaaat aggattgtag 20820

tctaattaca aagcgcccta tgggcgcttt tctttatcaa gtgatgttca taaaaagaaa 20880

ggggccgaat ggcccctttt tcttatgttg tattgatata cttatatttc atatcgcaaa 20940

aaggtggatg tttttttcgt caactcctgt tgtttcggcc tgcgcaaacc gttgcaggcc 21000

gtagttggta tgaatagttg cttattacat taattaggaa tcctattcaa aaaacatcaa 21060

ggaagcaaag taacatcaac aaacctgcca acaccatcaa cgttagttag ttgaactgta 21120

acagttgatg catctgcatt gacggaaact gcaactgtta caccagtagc acctgttact 21180

ccccacaatc cagtaggtga cgagaattcc tttccctgca gagtggatac agccgcagtc 21240

gcgctaattc ttcttataag aatttcaaca tcataagtac tgtaaactcc ggacgatgtc 21300

atagcttcaa tcagaatttt tctgcgcttt ggtcttggta ttgggtttga cgttgggtca 21360

gccaaagcag ggtgtgaaaa cgtaagtgta agtgtagagc cagcccctga tgacaaaacc 21420

tgatttgttt ttgccccacc aatattctgt cgtccagtgg tcatctggta caacccattt 21480

gttaccgtat cagaaacacc aaggttgttt ctgaaaagtt tattgtcaat ggtaatagat 21540

tctcgcaatg cagatactgt gtagccagtt aaatcgcacc cagtagcttg aaccgtcata 21600

tcagttgctg ttcctccggt ggtaatgaca cgttcaggtg tcaaggttcc agtaccgcca 21660

tcatcaaccc cttttgttgt cttaacatta gaaagcatca gatatgagcc acttccttcg 21720

atataaaagt gggtgttgta taggacgccc tcagcagaca gtctagcagg ccgctgtgag 21780

aataagttac cgataatagc tccaccacca tttctaatgg caaagtttgc agaaccgttc 21840

ctgtcaacaa gctctccggt tataatgttg cctacagatc cataaatatt atacccaacg 21900

ccatcgttcc actcgttacg cacgtttacg aatatgttgt tattcgcccc ttcttgtaaa 21960

tctacaccgt ggctagattg ggtatttatt actacatcaa taatctttga gtctataagg 22020

tttcttatgc caatagcatt ggcgcagatt gagcttctac ctacctgaat tgatgtatat 22080

ccataactgc cgacgccata caggaaacga tacaggccac agttatacag gatcccacca 22140

cgcaactgat ttttgttgtt gatgaatggt ctggccttgt ctcttccatc aaatatgcag 22200

ccgtaaattc ggaaaataga tccggggcca aagatatagt cggcccccaa agccttacgg 22260

attaccgtac ccttccctac gaaagaagag tcaccagaga ctgtgtatgg cttgctgcat 22320

tcgccgtaaa tcgtaaaacc tcctggcaaa attacagaac catttaaggt gtagatccct 22380

tttacggtat gggggaacct gacagatttg taaccagcat caatgatggc ttgcaatgcg 22440

taatcaacgg caacctctgc tcccggtgta ttaagcattg tattaatatc agcctttgtc 22500

attttgctga atatgtttcc ttctgctagt tgaattagcg tagcattacc aatattaacc 22560

ttgccaccac catcaggctg cgccagcgcc accagcgttg actgctcgcc agcgtcatta 22620

atgatctgct gaacttgatc tcttgcagtc tccgcagcca cctctgaagc cttggagtta 22680

gtctctgaag tttttgccgc attttcagaa gcggcagcag cagcagcact tgattccgca 22740

gcttcagagt ccgccttaat ttggttagca agattttgca gcgcttcgaa atcgaaatcc 22800

ttaaagaact cgacagcttc tgcaattacg gtttcttgcg actgatagta gcgcagagtt 22860

tccgcaacat cttgcgccag gccgtcaacg gttagcgagt cgcttaacag gatcgcgtaa 22920

tcgctagacg ctacgacagc gccttttgtg gtgatggctt taatttcagt atcgctaacc 22980

accttgttta cgacggccat ttgaatcggt gacgacaaaa acataatcgt cgcaccgggg 23040

cgaatcagcg aaagcgatga ttgccatttt gtgccagtcc cggtaacggt tccgtctgcg 23100

gccatagccg ccttgccttc tctgtatagt gccattttaa aagcctctta tgttggttga 23160

gtaacgcaga taatagcatc aatgaacaga taaaaaaagg agcctttcgg ctcctttagt 23220

tgtcaaatta gaacgggata tcatcatcga aatccatccc cgtattcccg ccgctgtttt 23280

gcggttttgg cggttgctga actttaggtt gttgaggctg gccccacccg gattgctgat 23340

tgccgccgtt ttgcgaaggc tcgcgctggc tgaattcgag ttgtggcata atcatttcat 23400

tgtggctgta aattgtgccg ttgtgctcgc ggttcacgat ctgaagcgtc cggcaggtga 23460

cgctgatcac cttatccatt tgcagcgctt cgtcgtacca cttaatcatg ttttctttgg 23520

caaagaaaac ggcgcggtag ttcgtgtaaa ccgtttcgtc ctggccatca cgattgcgga 23580

tcttcattcg ctccgacagg tctacggcgt acattttcca cggcccgtta ttgttactgc 23640

cttccttgac atacggttcc tttcggataa cacctgttac aacatgcatt gtcgttccta 23700

tggggcggtt tcccgcccgg ttaaattagt tgaaagatga aatatcttgc gcttctggtt 23760

caggctttga ttctaccttt tccggctccc gtttcgcaac ctcttgcggt ttaccagggt 23820

taaagccgtt agccggggtg actttcagtt ctgcctggcg tttggtgata tcgtcttccg 23880

tcattttcca ttccgcaggc gttaacgttt gtttcgccag cttatagatc tcacgaagcg 23940

attcgagatc ttcgcacgcg tcaatgcgtt ttttgaagtc tttcggcgtc attttcgtaa 24000

tttctgcatc atcatccgcc tgcttgatgc cgagagctgc ggccagtgca taacggcggg 24060

cgtaagatgt tgttgagccg tatgcttgtt cgacggtttt actgatcggc atattgtact 24120

gaaacgccat gaactcgccg ctttcgtgca aaaacatagt ttcgaggtgc ataacctttt 24180

cggtgctggt atccatcatg gattgaatga ccataatttt gttcttctcc agcgccgggg 24240

aaatagcgtc gaggatatca ccaagattcg cgtaggtgtt cccaagatgg ttgttcttcc 24300

cgcttttctt cgcggccacg aagccagatt ttgccttgat taatgcggct gcgatggtgg 24360

taaatttttc agatgtacgc atgataaagt ttccttttcc tgattggtaa tgcgcactat 24420

atcacaagtg cgcaccagtg tttagctatt tgtgccgtat acgtccggga acatgtattt 24480

cacaaactgc ggagtaggca aaacaacttc cgccgcgttt gactcatatg atggccatga 24540

atcgtgcttc acgcactcag catactgatg aatcacgctt tgatactgct tgcgaccgat 24600

ctcgatctgc tggctggtca gggtgaacgc cagcggagca aacggtgatt ttttctcctg 24660

cgttagaagt ctgacaacta ctgggcgttt ttcgttgtaa gtcttcacga acagatcgcg 24720

ctgcaatgcc atcttgagat agtagcccaa attgaaggcg agtcgcccga aatcgtccgg 24780

cttggaggat tgagtggttt tgtagtcggt aatcaccacg acctcgaaaa cctcatccgg 24840

gttgaacccc cactccttga ttagttctgg atcggaaaca acgtcaacat gatcgagtcg 24900

aaccttgacc ttgacgccga agatctcacc gaagattgac aattcacgct gtgcggtagg 24960

cgattcgata catgcggcgt gtcgcgggtt ggccaacatc acgcttcgca tttgaacaac 25020

ggcatcgaaa tcaacatcct taactagctt gcgcccggag ttcatcgcgg cgctttcgtc 25080

gcaaagttca atcgcccacc aaacatttac gtcaatcccg gcgcgatatg ccatttccag 25140

aagttccggg taatccttgt ttgacgtccc aatcaggcca cacgctttca gcttcgcaga 25200

cagtgccgac ttcgacgtaa tcagatcttt aacctcgccc ggagaagtcg cccgcaggta 25260

ctcgccatta aattttgctg tctcaagcat acatgtatgc gaacaggttc cgaacgccag 25320

cgcggctgtt tcctcacgca ctttgaattt ccagtgcgcc ggggatgttg cgtaaatctc 25380

gccgaggctt gagccgctaa cgtactccgc gcaccaggag ttaggatcgt gatattgctc 25440

gttagtcaat tcactgctgg tgtatgccct gaaaattgct tcagccattg atattgctcc 25500

atttgtggtt tcgttgcgtt aagtatacgc atgacgattc ccggcgcaag tcaaaaagtg 25560

ctatccgcag ttggttaaaa aacgagcgaa atttacgtaa gatttagtaa gatgcatctt 25620

acgtggtttt ttcaatacat ttcataaagt taacgcaaat cggtaagatg gtaagatccc 25680

tataggtaaa tatccatgaa aaatctggcg cgaaatcaag cgaaaaaaga cacgtacccc 25740

ggagaaatct taccaagatt gatagataca gatagagtaa taataataat attgttattt 25800

atcatatact tactatctat atattgctgt taattggtta aattttgcgc gaaatttacg 25860

taagattcat cttactaaat cttacctaaa gtggttcgac cagttgcaag ctactgaatt 25920

taaggcataa aaaaaggtaa gactgatttt ctcaatctta cctaatttct ggtaaatatt 25980

taatcagaga agagatgatg ccttgtataa cttacgctca aagccgagct taaagttgtc 26040

accgttcgga acgataactt taagatcccg atcgtcagca gccgccagca tatcccgatc 26100

tccacgacgg caaactaccc gcatttctcg cttaccttct ccgcccttgc atttatacct 26160

gtacgccacg atctcgacat tggaagggat gatgcaggcc cacacatcac acttgaatga 26220

actcgcaata ttgaagtgca tcgcctcaat ccaggagcga gcaaggtaaa tcggcccgtt 26280

accgtcgtcg ctctgattgg tcactatcac cgatccgaag gtcagatcgc cagctaacat 26340

cttctcgcgc ccctcctcat caatgaacag catgttgcaa tactcatcat ccagcccatc 26400

ttcatgcacg agttgcatcg gtagcgcgtg aattagctcc tgtctgccgt tctcgtgagt 26460

ttttactcca acctgatatg atttgatatg ctcattttca atgccctcat agagcgttac 26520

aggcgtgcta tcgacggcct ccgttctgtt taaaactgcc agcactcttt catgatctgc 26580

catcttgccg tagtcatacc cgttatcacg agctacctgc ttgtttctct tgaccacgta 26640

ttcttgcgga accttgccga gatagcgccc aagaatgttg atgcactcgc tatacggctg 26700

gccgcttaac ttcattaacc agccgatccc cttatcagca ccgcagccgc cacagtatgc 26760

gccgccgtcg cctcgcgttt ctaatttgtc agtccagcgg aatctgtctt tgccgccgca 26820

gttcgggcag tcctgatgct tgccgttgaa gtatcgagag tggatgccgc aaatgttctg 26880

caacgcttcg cgccacatgc cagccatgta cggcaaaacc tctttttcat cgtaaaaatc 26940

cacgtcgtca cctccaaata aaaaacgcct acacgagaat gataacccgg caggcgttta 27000

gtgtttagac aaattgtgct atcggactac gcggagcatt tcccggcggt cgcatcggcg 27060

cgtaactggc ttgccgttgc tgtcaaacct taaatctggt cggcagaatg aggcgcggaa 27120

acctttgcaa ttgttccggc ggtagctctt gtgtacgaga taagcgccat cggctgagat 27180

catgccgcgc ttacgccact gctgaacaac ctggatgcta acccccaact cttttgccgt 27240

tccagcgatg ccaccgaagg catcaataac aagctccatc cgcgcagtca acccggcgcg 27300

aacctcatcc ttcagcacgt agtaaccagt cggtcgcttg cgtttcttct tatctttccc 27360

gcgcgatgtt ccgttattgc cgttcaaggt tcgcttatcc acctttgcca tttgttccat 27420

aatttaaccc tcatagcatt tttttgttaa acatgataaa atgttccctg tattatacac 27480

gcaactatgc gaatgacaaa ttaggattgc ccatgctcac aattgaacaa caaattgaag 27540

cctacgcaga caagattccg ctaatacaaa agcggttcac cgtcggaaat atcgttcctt 27600

acccgtatca ggcggttgcg tatattgaga ccgcgaagcg gatcgcaaaa tatgaacatc 27660

ctttttacat taaggcttcg gtttccgccg ggaaaaccat catgatcgcc atgctcgcgg 27720

cgcagtgcaa ggcaatgaac ttacccatga tggttcttgc tcgccaggcc gagatcgtga 27780

agcaggattc cgaggagatc agtaacctcg atgttcccaa ctccgtttat tgcgccgggt 27840

taggcacgaa ggccgcatac ttcccgattg tcgtcggatc tgaagggacg gtggttaatg 27900

gcctgtttaa aatgcttggc gactacgtgc cttcagttct ggccattgac gaatgccacc 27960

aggttgactg gcaagatctg gcggaagcga tcgccaacaa tgaatcgttc gagtacatga 28020

gtagaccgaa ggataagccg tatcgcgtga acggggagct ggtcgatgcc gaccacccat 28080

acgacgaaaa attcgacgac gtggaattcg gcggcgggcg cactcaatac accatcgtaa 28140

tcattgagtt aatgcgccgt tgccttcaga agacggggcg cgagcttcgc attgttggct 28200

atacgggttc ggagtttcgc ggcgtcattc cgatcttaca ggaagacaag agccagccgg 28260

gattctggcg cgagcagatc accgacatta acacaaacta tcttgtcgag ttcggatcgg 28320

ttgttcccac cattttcggc gataccgagg cagatgggtt ggggtatgat ctttcagaat 28380

tccacggctc cagtcaggac ggcacgcagg attttagcgc ggaagacttg cgcaagatgg 28440

agaagaaaat ccatgaatcc ggcgaaatga cgaagctgat tatgcaaaag gtcgtggagc 28500

gtgcgaaaac cagaaacggc gtcctgataa cttgcgctgg acagcggcat tgcaaggagg 28560

cggcgagcta tctaccgccg gacgcaacat acgcgatcat caccgaaaag acgaactcaa 28620

agaaacgcgg agaaattttg gataaggcga atcgcgggga gattaaatac atcttccagg 28680

tgatggccct aaccactggc gttaacgttc cgttttggga tttttcggtg atattgcgca 28740

agatagggtc gcttacgttg cttattcagc ttttggggcg tggaatgcga cttctaaagg 28800

actggcaaaa gcagccgcct tactcgtggg tgaaagaaga tcaccttgtc tgggattttg 28860

ccggaacaat ggatgacctt ggccagttgt atttcgatcc gattttggag caggcgcaat 28920

accaaagacg taagagcagc aagaacggcc cgaaaatttg cccggtatgc aagggagaaa 28980

atagcgagta cgcccgccga tgcatccaca aagacagcaa cggtaatcgt tgcgaatact 29040

tctggatctc gcagcgctgc gaagaccaga aagacccgcg aacagggaag attaaagtaa 29100

aggggtgtta cgctgaaaac gatattgttg ctcgccagtg cagatgctgt ggggtgcagc 29160

ttaaagaccc caatgacaat ctcaccggga agcactacac gcaaaatgac tggtatgatg 29220

ttgtcgggtt cgatatcgga ttgactcgca atcagtccgg gatcatcttc aattacgtgc 29280

tactgaacca tgacggcgag cggttcaccg caagggagaa gttctttccg gaatcagaga 29340

atcagatttg cggcaagtta tggcggcaaa aggcagtctt ccagcacgtt gacgacgcgg 29400

taatgcgcgg caagttgggc gggatgaaaa atgcgcgaaa aatccttgag agtgcgcatt 29460

acttccgagc gccgaagcgc gtaacgcatc gcgttaacgg taagaaggaa gatattattt 29520

cacgcaaaga ttttggagac gcagaatgat cacagataaa ggtgattatc tcgaatacta 29580

cggcgggcct gtaaaggctt gcccgcttga gaaaatagac cagatgaaca gcgtttcgtg 29640

gcttcggcac gaataccctg attatctgtt ttggcatacg gttaatgagg gaagcaagca 29700

caaagcgagc gcggttatcg atcaccaaat ggggttgttg aagggtgtta gcgacattct 29760

catcctgatt ggtttcggtg gcaaataccc gttcgcagcc attgagctaa agcgccaggg 29820

taaggcgcag gcttcgccag tgagcaagga gcaaagggaa ttccttgctg ccgtccggcg 29880

tcgtggcgga ttcgccgccg tggcctatgg cttcgagcaa ttcaagatcg ctttctgcga 29940

tgccatcaaa tagcactttt tgttaaaact gcccggcgaa agccgggtat catttcccca 30000

tcgaaactaa gaatggagtg ttgaaaatga aaaagatgct ggctttagtt gttctgtctc 30060

ttggtcttat tggttgcagc gaaaaaccga aaacatatga ttgcggtggt gaagcgtttg 30120

aggtaactag caactatatg aaagttgtca agggtgagaa ctctggcgtt ataattgatg 30180

gcgctggcga aaatcaatat aaactgctta ctccttttgg gtacgctcat tatgcagtca 30240

acaaaaacac tattgatgtt agtgttggcg cttttcacaa taccttaacc tgcgaggtta 30300

aataataatg gcaaaagata tcgcagacaa agaaaatcat gacgcgttca ccacatttga 30360

gcagattgag cgagaaacgt tcatcggtaa cgctcttgcc actggtggac actatcaggc 30420

cgtcaggccc gacaagtttt accaggtaac aggcaaccga tacgtcggga gcaaaacgcc 30480

ggatattgtg cgagataagt gggcaaccga tcgcagcctg atcgcataca tggaagagcg 30540

ttacggccct tacgatcttg acgccgccgc agaccaaagc aacgcagttt gcccgaagtt 30600

ctacgacgaa aaaacggatt gccttaaacg ctggtggggt aaaaacaagc acgtatggct 30660

gaatccgcct tactcgtttc cagatccgtt cgtgctcaaa gccattgagc aaatggagca 30720

cgataaccag atcgacattc tgctacccgg cgacaattct acggcctggt tccgtgacgc 30780

gcagaagatg gccgccgaaa ttatctggat tgttgccgat gttgaagagg atgaggacgg 30840

gaaccagtta agccgatccg gtcgcctcgc attcatcaat ggattaagcg ggaagccagt 30900

cgacaacaac aataaaggaa gtgttatttt catcatgcgc aagctcaagc cgggagagga 30960

gcaaaagacg ctttacattc cggtaagcga gatttgcccg tcattagcta aaaagcgtat 31020

gcgcaaacgt gggatctgaa aaatggaaca ggtagaatct ttctccgagt atcttcggat 31080

tgtggttgaa ttgctggaca agtacggctt cattggtacg gatgaggaaa agttagcctt 31140

cgctgacacc attgacggaa cctacatgga gttcatggac aacggaaccc cggtcgctga 31200

ctggccagaa attcttgaac gagaattgat tgagtttaaa tcgcatgaag gcgctgagta 31260

tttcgcaaaa cagcactaat tgctaaataa tgcccgccgc gtgcgggtat tattactcca 31320

tcaaccaatc aggagcaaac gccatgaaaa ccaaaaccat tgcagatact atcaagatcg 31380

caccagcaaa agcgcaagtt gtatcgcgcc acctggttaa catgtctcgc ctgtgcatgg 31440

ccgactacat ggcgaaccct tcagagaatg gccttgatgg tgcaattggc gagatttatt 31500

ttcgcgccgg gtacggcctg gaaagtgtgg ccatgtatga gcaaatggcc gagggttttt 31560

gcatttacgg tgacgaatga tgattgtaga gactggtcgc gctgccgtat ggcagcacgc 31620

aaaagaggct ggaataagtg atgatatcgt gaagatcgca aagtatttcg atatcaaaga 31680

tatatcaatt atctttggtg ggaagctcac ctatctacac gaacgccctg tgaagcgcac 31740

gcgaatagca gtggcaacgc gagcggaggc ggacgcgctg aagatgttca tccacgagtc 31800

gaagcagcag aagaaatatt acaagtagcg ggagtgtcga gaatgcgata tattgcgatc 31860

ttctttactg cgatcctgtt cacgatcgca atcattaact atgcaattca atttggataa 31920

attatgtcac ctaaaatcac agacgaagaa tttttagccg cccgcgagga aggcaaaacc 31980

taccgcgaga tcgcggaaga gttcggcatg aacattcgaa gcgttgaacg tcgcggagtt 32040

cgcctggcgc gacaaggaca cctacacgga aacgcccacg ttgcgaagca tatcccggac 32100

ggcttcggcg tcaaaggcac gtcgacgatg attcgcgcgg acggctccga ggtcgttcgg 32160

tgggttaagt cggaagtaga ccgcgatcgc atggttgcgc ttatggaggc ggcgcaggca 32220

gccttctgcg aagaccttcc gcgagccgaa ccgcaaccgc tggatgagtc gaagttctac 32280

attgaagatc agcttgcact gtacccgatc ttcgacctgc atattggggc aatggcgcat 32340

aagcatgaat gcggcgagaa ctacgatacc agcacggcag agaaggttct aaaccgcttc 32400

tttgattatt ccgtttcggt ggctccgcaa tcacaaaagg cggttttgtt ggtcggtggt 32460

gacttccttc acagtgacgg actggacgca gtaaccccgg caagcggtca cgttctcgat 32520

caggacagcc gatacgcaaa acttgtttat gtggccattc gctcgctgcg tcgcgccgtg 32580

tcgctactgc ttaacaatca tgcggaagtt gaagtgcagg tgattgaagg caaccacgac 32640

caggccggga tgatttggct acgcgcagcg ttggcggcgt tctatgagaa tgaaccgcgc 32700

gttttcgttg atgttagtcc ggcgatcctg catcgcacct tgtggggcaa aaccatgctg 32760

ggctatacgc atggccacac gatgaaaaag ccggaaacgc gccttgctgc gatggctacc 32820

gacttccgta aggagttcgg ccagtgcgac tacatttaca cgcattccgg ccactggcat 32880

caccagactg taacggaaca ctcgttaggc attgacgaag tgcatggcca gttaggcgca 32940

aaagatgcct acgccgcacg cggcggatgg cgttcatacc gccaggctgc ggtgattctg 33000

tacagcaaag aatatggcga agtaggccgc tttatctacc gtccgaacat gtaaccacaa 33060

cggccccgcg aggggccaac aaggaaaacc gatgaataga aatatctgca ttttcgatct 33120

cgatggcacg ctttccgacg gaacccaccg cttgcacctg ctgccgaaaa aagatctcca 33180

ccttacagaa agttggagcg aatttaatgg cgcgtcaatt ggcgacagcc caatccaaag 33240

cactattgac gtggcgaatg cgctttatcg atccggaatg accgttatca tcctgactgg 33300

ccgatccgat gaggtgaaga ccgaaacaat gatttggctt gaccgctacg gggtgaaata 33360

tgacagccta atcatgcgcc gcgccagcga taaccgtaaa gacacggtaa tcaaggagga 33420

ggagttacgc aaaatcggac ttgatcgcat tgttgcagcg tgggatgatt cccccaatgt 33480

tattgcgcac ttgcgcggcc tggggatcac gacttaccag gtctgcgact acggcgacaa 33540

tcttcacgat catttaaaat cacacggagt agacaaatga aaaatgtaat tatcctcaac 33600

ggagcgccgg gcatcggaaa ggacactatc gcggaaatca tctcgcggaa gtggcaatac 33660

aagaacctta gcttcaaaca gccgatgttt gccattgctc gtgctgtgct gggatcggct 33720

gattttgcac gctttactgc ccgataccac gaccgcaagc acaaagaagt gaaatgcgat 33780

tttttgggcg accgttctcc gcgtgaattc atgattcaca ttagcgaaaa tttcgtcaag 33840

ccgaccctgg gcaaaaatca gttcggcaag ttgctttgcg attcggcgct aacttcgccg 33900

tttaactgca tcgtcagcga cggcggattc gatgaggagg tggagcacgt cgcagcgcat 33960

gaggcgctta acgtgtttgt cgtccgcctt catcgtgacg gcatgacctt tgagggtgat 34020

agccgaaagc atattcgacg cccggatctt atttgcgaca cttaccacga actcgatttt 34080

gatatgacca ctggcgagcc ggaagacgac gcgcaaaaaa tccttgatat ggtgtcagat 34140

gttgcattaa aattataaag ttaatgcctt tattatcatc accttaacta ttgggaacct 34200

tgatgggttc cctttttttt gttcttaatt tggcctaatg catatatcat caccttacca 34260

tttaactaac agaggttgca tatcatgcga gaattcatca acgcggcaac caatagcagc 34320

ggtggcgttg ccctcgcggg atctgcaacc gggcaattaa tcattgctgc cattggctta 34380

tttttcatgg ttctattcgg ctccttcggc gcgtggttgc gctggcgaga ttcaaaggcg 34440

cttcgtgaag cgctggaagc cggggatatc aaaacggcgg tgaagatcag gagtaaataa 34500

catggggatt aaaacgcggg ttacattcgc ggcggcggtg gcgatcgcgg tcgcgttcct 34560

ccccgaagtg gaggacacga aatacaatgt ttatatggat atcgctggcg tcccgacagt 34620

atgcgaaggc atcacaggcc cggacgttat caaaggcaaa acctataccc ggtcagagtg 34680

cgacgcgctt ttaaccaagc atatccaggt ggcgaagcga accgttgaca gcaaaatcaa 34740

agtcgatgtt ccggacacct tcagggcgtc gatgtacagc tttacgttca acgctggcgg 34800

cggcgcatat tctggcagca ccatgctgaa attaacgaac catggccgat tgcgcgaggc 34860

gtgcgagcag ctatatcgct ggacgtacta ccgcaacccg aagacgggta aaatggagaa 34920

gtcaaaaggc ttgtataatc gccgggttca ggaatatcaa ctatgcatta aggatctgaa 34980

atgagcacat taaactttca acgagcgctg gccatcggct ttatcgtgtg ggcggctgcc 35040

gtcgtttccg gttgcgcgtc aagcgtccca atcctttccg atctggttgg tagcaagccg 35100

gatatgacgg cgcaagtcgg cgcggagaac gttaaacagg cggttggcgt gaccaacaaa 35160

acggatacgt cgagcaagca ggagaccacg ttcaaagagt cggcggtagg taaggttgat 35220

acgtcgaaca agaaatcggt gacgacctcc agcattcacg ccaaccagat tacggcggac 35280

aagatcgaga tccggaatga tgaaagcgga agcctgattc cgtggctgat tggtggtgtt 35340

ggggtagtaa tgctggcgat cggggtgttc agcctttggc gggaaagaaa aaacaaaggg 35400

gcgtaatgcc cctttttcta tatgtaccgc ttgacgtgca atagcgctac tccgtcttca 35460

tcgttaagcc catgctcaac cgtgttggtt gcggccattc cttgatagag caagatgagc 35520

gcggcacgca agtaattttc tggtgtgatc tgtttcacgc aaacaagcct gtgaacctcc 35580

gttatcagat cttccacctg gtttcccgaa aagctgttca tcactgagtt ggtcaaggtg 35640

catcatctcc cacatatatc tgttatccat cccatcgaac gaccggaaat caaaacctat 35700

ttccctgtta tccggccccg tacaccacac agcgccgttt tttccgtcaa ggtatccatt 35760

cgtatagctt cgcgctaaaa actccttaga gaccattgag gcgaacatac gttgcgatac 35820

gtttgcggct tttgccaggc gaggcgcttc gcggtgcata tagacgaact tcgcaaaatc 35880

ctgccgggtg aattcccggc tagattcgca gaacttgtaa atgtcaagaa tgaacatcaa 35940

ttacctcatg aacgacgggt tgataaagac tttggaatcc atcacgcaga taaagcctaa 36000

ttcctccatc ttcggcaata agcgttcctt aatctttttg ctaaccccgg cctggccttt 36060

gaagatcttc aggttgcggc aggcgttata aatgccctgg acggtcataa cacctttcgc 36120

ctgtttgcaa cgactggcga taacgtcata cagcgcttta atttccgctc cctcaccagc 36180

aaagccggaa gagtcagccg acgacaaata cgttttgctc aattcatgga acatgatgat 36240

cgcttcgtca atggtcgccg tgtcgatctt cttcgagcgc ttcccgcctg gttgccagtt 36300

ccggatcgtg tgaatcacgg acgccagacg cataacctgc ttatcaaact tacccatcgc 36360

gccgcgaagc attgtatgcg aatacttacc gccgtcgcct aactccggtt ctaattcctg 36420

gcgggccttg ttcagtctcc gcattgccgc gtcagtaacc tgcaacttaa cgtttgattc 36480

gctcataatg tcgtgaatca gccggaagta atccgccttc agtgactggt cgatcggctc 36540

ataggtcgaa ttcccgtttt cgtcgatgaa cacgcgctca cccaaacggg tttgctcacg 36600

aaccaaaagg aaacgctccg agacaccgat cccgcgagaa ccagcctgca taatggcgtc 36660

gatggtttca tcctgtgcaa ttacgcaaat gcagcccaaa gccacgaatg acatattatt 36720

gctaacgtcg gcacgagcga tcgatacgtg gcctttatcc catgctttga gcaccagttc 36780

gctgttcgtc ttcttgccgc catcgttgcc atacgtgatc cccaaaaggc tgttaaccgc 36840

cgtcgcctca tcggaaataa cggcaaagtt tccctggcgg ttgttaatct tcgccaggcc 36900

ttccggggtg gtatcggata ccgggaaagt tagatcacat aatttttcaa gtttctcttc 36960

aagttcgtca cgatcttcga aaagtttcac catatcagat tgcgacaact cacctttaag 37020

cgcctgcttg ttggcggaca acttcgccat gattttctta cgctctttct tgcgcgactc 37080

gttaatgcgc tcgacttcgg cgacgatcgg atcgatggcc agcgagttaa tggcagactt 37140

accagcggaa ggcggctgcg atgtgataac gtaaagcgtt gtcggttgct cgcttccgtg 37200

gtactcgacc caaaatcgac ccatcatcgc ggcggacacg gttccgagaa agtgcatgta 37260

agcggatgat tccgggaact gaacagaacg tgctgcattg agcgccagct tgccgaccac 37320

atcgtaatca ttggcgatcg agattgttgg gtacttatcc gcgtttacgt cgatatcttt 37380

gggctttggc cagaatgaaa cggagtcacg atacccgttc gcacgaatcg caacacgcag 37440

aggactgata ccctcccttt ctgcgatggc gataatatct tgcggtgata cgcggtcatt 37500

taaaaacatg cccttgctcc tgattggtta atcgttcggc taatcatatg ccgtattaaa 37560

tccgagatcc agtgataatc taaacacgct tgaaaaacgc gcttggatta tcggggcgca 37620

tggccccgaa cccgtcacag gtatttagct tcgaaagtcg ttccgtccga tacactgaag 37680

ccaacttcct cgcggtacag cgtccagcgg catccgtcac ggtcgaagat gtatccagca 37740

accgcaccga gcgcacgacc gctttcgacc tggtagcgct tgccgacgct gaaggatttt 37800

ttcatcgggt tgctatggtc aaggccgacg cattttagcg ttttggtttt gagttcgatg 37860

aacgtcgcaa ccgccgctcc gccgtcgcca gcaataaaca actcaccgct aacgcctacc 37920

gacaaaataa cgcgcttctt cttcagttcc acgctgtcat ataccatcat tgacacgttg 37980

ccttcatcgt cgacgcgggc ggaataaagg ttgttttcat ggatgttaac ggcacggctt 38040

gacgtgcatt taatcttgat tgacttcgcc atttattttc tttctcccat ctcgatatac 38100

aggttaacca gatccagaaa gtcagcttta tttctgcaac tcaatttaaa gccaacgtgc 38160

aactcaattt tattttgcat cgcatccagg gttgcgccgc cattattcat cctgagaact 38220

tcgcggcata cttctgcgag tttttgagat atcatttctt caccaccttt atttccatgc 38280

cttcgcaagc ggccttgata tcattaatca gacgattaag ccttaaatca gtcattacat 38340

ggtgatgacg aacatcaata atcccggcaa ctactaacag catgacggca aagataaaac 38400

caccaggccc ggacatacac aaaatagtta acatcatcag gataataaat ttcatttccg 38460

tttactccgt ttcgtttcga tgtggtaatg ctacccgact ttcgccgggt agttttagca 38520

attcgtgcta ttctcgataa ttctcctgga acacggcgcg ggcaaaacct cgcggagtaa 38580

tggagcgtag cattttagtc ctttctgacc ttccgcccaa aaacttccag gcccaaaaga 38640

aattcacgcc ttctactcca tccggcggcg ggagccgttt cggttcgaca aaaccgtttc 38700

cgtgccagat gcacgttttc ttcgtgtagt tgtcgcagtg tggcattttg ggatgccaga 38760

caggctcatt tggtgataca tagccgccga agtcgcgcgg gtgaaaataa aagtctggct 38820

tacgccatag tgtcgacaat ttgccaaccg ggttttcgac catccagggg caaccatact 38880

cattgcccaa ttgctcgacc atcttcgcat catcggcggc tgatagaaca tcctgaatat 38940

tccgggcgtg ctttacccca ctatgcgcta acagcgtgca ggacgggaac gcgaaaatga 39000

agtcaggatc tggaattccg agaatcgatc gcttcacgtc aaaatctttg tcaatccaga 39060

tattaacata ctggatattt gggtgaacca tgcggatatt gtattcgccg tggtcgccgg 39120

agtctgcatt gaaacagtac accttgcatc ccttaatggc ccagggcagg cccatgatgc 39180

cggagccgtc gaacatgcag taaataacct tgtctttcat catcacctca ccagaaaggg 39240

acgcattcgc tgcaaccttc gaagcaggag cagccgcacg tattctcttc ttcatccgcc 39300

aggtcgtcgc acatcggcag gctttcggcg ttcatatcca tatcaccaag cgcatcagca 39360

agcgtcattt cctgattggc aaccatgatg caggcttcac ggttaaggcc agcgtcctga 39420

gccgcaagga agcgactcca gtatgcatcg tcgtaattat cagccattac tttttctccc 39480

aatcatgcac gccagctttc gataaacatg ctattcgatt ccagcgtcag cgtgacgcca 39540

gccagggagg actgaacgag gcgaccgccc ggaaccgtcc cgatgatgta acggcccttc 39600

gacttcgtga tcgtgatctg agtgaagccg gaggcgcggt tcatgcggag aacgacgcgg 39660

ccatgtgcgt taaggacggc gagaatggaa tcagtcttcg gatggttcat tgtgtagctc 39720

cttcgcttgt tggtgtgggg atagtatgcc actatcccca aattctgttt tagcaattcg 39780

tgctattcgg gaacgcaggt cgtgtaatcc tggcagatgt agattgtgcg ctgaacacgc 39840

ggcgcggcat cctccaggct ctgaatctcg atatcgttgc tgtcgacccg gcagcgtgtt 39900

gcaccgaaga tgaaataccc cgtctgcttg tcgttgattt gccacgcctt gctttcggga 39960

accgtattga cgacaaacgg cgtgtcgtcg gctttgaagt cttcagggtg acgcccgtaa 40020

tagtggatga ttgcagcagt tgccgccgcc ttaaactttt ccgtcacgtc acgggcggcg 40080

atctcacgcg agatctggat atcaacatcc ggcgcgccga ccatgtagga aaagctgttc 40140

gaaatggtaa ctgttcgcat tatattttaa cctcgttttg gtggatgaag tcggcgacgt 40200

aatggcattc gatcgggttc tcgatacccg cgagcacctc gcacgacccg ttattcagta 40260

gctggcaatg ctggcactct gcaccgccaa cactggcaat gacaatgacg atcccgttgc 40320

atgaaccgcc gaacattctg tgcgggaagg ggtaagcatc gcaggtgcaa accacctccc 40380

cggcccgccg tttcctcata cctggccctt ctcgatgcag aaaacccaat cacgagcgtc 40440

aaccgcaaag cgtgcaacgc cgtcgagcac cacgaagaca gtaaatccat tgacgccgcg 40500

ccccttctcg atatcggttg cggtacggaa cagatcgctt gcgccatatt gaattttatc 40560

cccgacgact acatccgcga atgctttcat gttgattctc ctgactggtt tcgatgggtt 40620

aactatacca ggcctcgcgg cccggcgttt agcaattcgt gcttatttca ctgctttctt 40680

cgctgccttg tacgcttctt tgaaggccgg aacatcttcg accggaatcc agaagccgga 40740

gccatagcca tcatcatagc aagggcattg gtcgcagtag tcgccccagg ttttctcttc 40800

gccgccgaca gagaagccag tggccatttc gcataatgct tcctcaaccg cttcgatctt 40860

gtctgcgtcg ctatccatga taacgaagtt ccacttgccg ttatactcgc tgcccaggtt 40920

gattgattcg cgttgaagtt tcatagcttt agcccctttg gctcaattcg tttcgttgaa 40980

gtcatcatag caaagtcgat cgggtgggtt ttgacaaaaa atgctattcc tgcgattgct 41040

gattattcca ccgattagga atagtggaat agatggtgat tgcacctgat tccccggtga 41100

ttagttttga cgaatcatga ttgcttttga ttgtttcatt ctgcgcaacg attttgggag 41160

tgtttcagga tttgcaacga attgcataaa tgtatgcacc atgcacgcaa tttgccgatc 41220

gaagccagca aaacgttggg tgtgtttcgc caacatttac gtcgtgtggg aagttgggga 41280

agattgaacc ttacctaaaa tttgcgcaaa aaatgaccaa tataagccca tatatagata 41340

gtaagtatat gataaataat aatattatta ttattactct ttatctttat atatgtttgg 41400

taagatttct ccgggggggg gggggttatg gattcctctt gcgcaagcca taaattttta 41460

gatgttccgc cgcgaaaaat gtaaataggt atatatagtg gatgacagca aatcttacca 41520

aacacgccga aaccctttga cggcaaggcc tgcggcctgt attatcatct tccgaaacat 41580

tacaaaacgc cgtagatgtg ggggaaacac aaatgcaata taaaggctat gagttcgaag 41640

aagtcgacgt gtccgagtgg atgcggtacg acgcagataa gcaggccgct taccatgaat 41700

ggctgcaatc catcacgttt ggtgacacca ccaggccagc aggacggagc caggaagcct 41760

cgaagccgga aggtacattc actggcaacc gctcttccga gatccggcgc atgttcctgg 41820

acggcgggcg gcgtatccag atgacggcgg aggagttcgc ggagaagttc ggcgttcatc 41880

cggtcgacaa ccatttccgc aggccgttgc tgaaacttct ggagccgggc gaggtgttgc 41940

gagtgagcct gggggcgggg ttaatctccg tctttaccga gttcgacgca atgactggct 42000

caaacctgga ggctgaggca tacaggcggc agggcgctat ctctgagcgg gaaagaatcc 42060

tttccatcat caacgaagta atgcctgaag gcatcatgct aaaaggctat aagctggcac 42120

taatcgccaa aatcacagga gaataaccat gaaactatat ccagaccaaa acgaccccgc 42180

cgcaccgatg cgccttggcg acacgccaat cgacgagcag gaggttatcg aagccattgc 42240

cactgcatcg gacgacgaaa atccaaccgc cctggaatgc gttacagccg acttgctgtt 42300

aatgtcattt cttccggaat ctgaatctta cattcagcac atcaccgaga cgggtcaggc 42360

ggtgacgaag ccggaattca tcggtgcgtc gaacgtcctg gcgatgatgg ccgatgaggc 42420

cgacaacatc attagcctgg attatccaca atgagcgcct ttatcctgtt ttccatcggt 42480

gttaattgcg gattgctcct ggcggcattt atcatccacc tgagcaacaa gtagcacttt 42540

ttgttaaaac ccggtcagag cgatcgggta ttatctcttc atcgacaacg aaccgaggaa 42600

agcgaaatga ttactctgat tacctgggaa cacgaaaaca gcaagccgga agtacgcgag 42660

ttcgagaccg tggcggcgtg ctacaacctg gcggcaaacg gtggctttta caaggcgcag 42720

attgttaacg agttcggggt tgtcgactat gaattttaag gcgagcgatg taaaacctgg 42780

cgcggtctac ggatcgcgct tcaacgaccg cctttggcgc tgggatggcg aaactatgtg 42840

gacgaaaggc gaattcgacg tgatttggca tgaatgcggc tggccccatc cgacgatgga 42900

gcgccgggat atcgcttact atctttcaat cggttatatg catgaggtgg tcgatagcac 42960

tttttgctaa aacaccgatg gcgagatccg gtattattta ttcatcgaca acgaaccgag 43020

gtgatgaaga tggataaggt tatcaagatg agcgaagtta agccgggcat gatggttaag 43080

tttgctggca agtttcgtct ggttctcgca gcagatcgca aggcgaatat tctcactatt 43140

cgcgttaatg gaaaggccca actattcgcg ccgcagtccg acattgaagt tgaagttcgt 43200

atcaagtaag caacctggag atgaagatga gcaagaagaa tccatattca accagcaatt 43260

accataacga acacgtcatg ctctggcact ggcagaaact caaagagcgc caccttgcac 43320

gcaagggcaa ataatcgcaa tagcgccgct tcggtggcgc tgtattccaa ccaccagcac 43380

ggcaaacgcc atgaggagtt gtcatgccac gattttcagc aacaaccaaa cttcgcacct 43440

tcgccgggat gctgatccca tactcatcaa ccaaggccgt ccagggcagc gagcacggcg 43500

tatatttcca ctggttaggt aagtggagat tcaccgtcat tcgcggcttt tacgtgacct 43560

gcgaccgcgt ggatatcgaa gaccattccg gcggaaacca ggttcacgag ttcaaaaagc 43620

acgaatagca ctttttgcta aaactcgatc ggggcaatgc gttattatta ccccatcgaa 43680

acagaggagc aaaaacatga ttggcaacca caacaacgaa ctgaacgcag cagcccaccg 43740

tcgcgccgtc gagcagaact tcaacgcgct taaagtagcc tgcgatgaga tgaaagcaat 43800

gcttgaactg ccgtcatggg atccccaact agaagactac tatgacggcc tccgcgtcaa 43860

gcgtgacgat atcatcaacc gcctgcgcct ggccggaatg ttcctgtaag gagaacgcaa 43920

tgattcacga agttaagtcc ggcgagaaag tagtggccac catcacgcag cgccacgttg 43980

tagccttcca gattaacatc cctggcacgt tcgatatgca gtcgcttgac gtgccagttt 44040

gggccaacac ggttgccatt gatgccgatg gctcaatttg ggcctatgag tcgaccgttg 44100

aaaatgttcg cctactgagc tactcgcctg attcatgggt gtataagggg ccgggcggaa 44160

acagaatggt tcaggtgggc gaaatggctc gcttccctga ttgggcaaaa tcgaagattg 44220

accttcgcgg cctgaaatag cacgcattgc taaacgcccg gtcagaggtc gggcataatt 44280

accacgaagt cgattaacga cccaattatg caaggagaca ggataatgca gcaattcaaa 44340

tctcgcggca gggtatacag cctgccagat accgcaactc atgccgcgcc cggcgtatgc 44400

cacggcctat acttcatgga tggcggcaag tggttcttca ttggcgatat ggtaacggat 44460

ggcgtgcagc cgctcctttg cggcaaacat cgcggattct atgatcacga tatcgttgag 44520

ttggagaaga agcgcgatcc gttcggtttt tggaacaaaa tcaaaggggc gatctttaaa 44580

tgaaccaatt cgaattcctg gcgcgggcgc gccgtgcgct tgcaatgtac gccttcagca 44640

agaacatgaa gattatcagt gttgaggttg agttcaacaa tatcagcatt tccggctatg 44700

tgggcggctc attacatgta acaaaattca gctttccgtc gctggccagc ctggagcgtg 44760

aggcgttcgg tttctgaata gcactttttg ctaaacgccc gaccagaaga tcgggcataa 44820

tggccacata ccaaacaacg gagatttacc atgattcgca ctaccaggcc gctacccaat 44880

aacctttacg ccacgattgg gattcgcgcg gtgagactga ttcacgtcaa cgaagttggc 44940

aacgtggcta ttttgtggtt taagcgccac tcaagcgggg ttggcctgaa cattgaaaga 45000

ttcgggcgct ttaacggcaa ctcccaaacc cattctcatt gggcggatgg aacttaccac 45060

tggacgcacg ttaagcacgt caagaacttc gacgccaaat cactgagcga cgacgacgcc 45120

tggaagatgg taacagatct cttcgggtat gatgcctgca tggaggtagt tggatgatac 45180

gtttatcaga cttcgatcgc cactgcttaa ccgggcaatt tggcgaaaag ccagtgttgt 45240

gcaagatctc gaaggttaag ggcgagccgt ttgagcaaat caggacgctt ataagttttg 45300

ccagacgaaa ccgcctttac atccaggcac gcggaggact gttcgcaaag caggttcatt 45360

ttgcgtatgg aacagggttc tacactggcg gagacggggt taacccatca ccgaaaaggg 45420

ttaggctgcg cgagataatc gaacgcggtt acgtttggac taacgggatc tactaatgga 45480

aacggttaaa gtaatttgca cgcacgtcgg ctattcgccg ctatcaacct atttcacccc 45540

tggccgcgag tatgacgcca ggttcggccc tggccttgat gaagtgtgga ttttgcagga 45600

cgaccaggga accacggagg acgaagaatt ctggccagcc tgccgtatgc cggacgaaaa 45660

gatcgcaatg tacgccaata ggccatacga aaacaacgct ttatttgagg tgaaagtatg 45720

aaagtaagca aaaagaaact cgcgctgtat tgggccttct tcctggtcgg cctggtactg 45780

catgaccacg taggctggca ggcgatcggc atgatttgca tcatcattgg ggtatgccgc 45840

ctgtctgaga ttaacgggtt ccgccgtggg tatcgggcag cgttcagcgg cgaaaaagaa 45900

tagcactttt tgttaaaact gccgtaacgt catttgataa agtggcgtta ctgaagtgaa 45960

caatccaatc aggagcaacg caatgaacga caatttaatc gcactgctgg aagaactgaa 46020

ttcctacggc tgcgcataca tcgagctttc gagcgagcag gttattgaag tcgcaatcga 46080

cgatgacgcc gggttcacca gattaagggt ggcggcaggg gccgaagagg aattcgagga 46140

ctgcggcatc cgtgaagtaa ccaaccttct cgaatcccac caggtgaagg agatccgata 46200

atgaaaaccg tcaaacttat ggttgtcaat gcggcccgcg aacgtgaacg ccaccttatc 46260

ggctgcgtat tcgatgccgt cgaggtgcac gaaccgctag gcacagtgca cgtaatccag 46320

ggcggatacc tgaacgaatt catcgtcaac ccgctggaca ccgacgaaga cttgaccatc 46380

cgtgttggtg gcgagccgga aaacgcgctt ttccacctgc ggaaagtatc gaaggcgtcg 46440

cggcgcaagt tgatggttcg cgcactgaag cgcatagcga agcagcgcgg atacaccgac 46500

gcatacatcc gcaagttcgg cacgttcgct ggcgtaggcg attgggcgcg gtcatgggcc 46560

gacttttact tcaacaaatc cggcgaattc tgtgccgcgc atacggactg gctgcatgga 46620

accgaacacg atcccgttaa cggccacgag tggtacgaat tcattgaggc ggaattagac 46680

gccctttgat agcacgaatt gctaaaacga acgtccgggg atggtgcata ctatccccac 46740

actaacaaga gatggagcct aaccatgaac accacacaaa tcatacacct gatcccgtcc 46800

gacaacttcg ccaaagacga cgacccgctg ttagccgccg tcattcgcgc cgggatcgat 46860

tgggcgctga gtgatgcatc gtgcgaatca ttcatgcgtg aaccattctc acgaatcgcg 46920

gaacttgaaa agcaggttgc tttcctgact ggtgaacttc aggcggcgca caccaagatc 46980

gagaaggttc gggaaatcat gcgcagcgaa cctggctatg acgttgaaga tcacgcgcga 47040

gttttgcgca tgatggccga cgcattcgca aaactccagc aatagcacga accacaaaca 47100

ctaaccgggg ttgctgcatg gctaccccac caaagacaag gaaaaacatc atgatttacg 47160

caattattgc cctggctgtt cttgttctgg ccctgtatgc gtgtggctgt ttactcatgc 47220

gagccttcct gaaatctgct gattccaatg acaaagacga tctttatcct gtgttgtgct 47280

ggccctggat tacactttcc gccgttggca gcgtgatcat ttcccgcaat tttaagtggt 47340

gagatggcgc ttgggtgcca aaatggatag cctgccaaat agcacgaatt gctaaaactt 47400

acgcaaggcc atttgatata gtggccttat tgaagcgaac caatcaggag ctaaacaaat 47460

gagcattccg tatccgacca aagacaaatc attcctgggc ctggccgttg ttgagattat 47520

caaccctgac gttaccacct cccaggatga gcctaccctt gcacagtggg ccggattcaa 47580

agcaggtgat cgactgaccg tcgaatgcta tagcgacggc gagaagtggg cgcaggtagg 47640

gcgcgactac cacaccgaag aattcggcca tgttctccct accgactggt tcaaacttaa 47700

cgaaggcgaa tataaggtga tcgaagaatg aacaacaaaa tttgggtgct gacctacact 47760

atcggaacta acgaggggcg caaatcgcgc cgcctcacct gcgacacgaa agcacaggcg 47820

gaaatgcagc aggccgtgct gggcggcgag gtggccgaat acatccgcca gccagaatcc 47880

ttccatgtca attggcctga gaagatggat gttgacgccg tgctgcatga aatgcgccag 47940

gtgcaaaacg accaggcggc gtggaaagac ctgtaccttt gcggcgacgc tgagtcagta 48000

ctggatcact tccgctttgt ccggcaggcg cacgcggaat ggtcagatcg ccagtttggc 48060

aacgttgggc cagtcgggcc actcaaacac ctggcgaaag aggcgatcga agccgccgaa 48120

gcgccggatg acattagcga gttcgccgat atcctgatgt tggtatggga cgcaacgcgc 48180

cgcgccggat tcagtgatga gcagttggtt gaagcggtgg cggaaaagct ggagcggaac 48240

aagcgccggg cgtggagcga ggtcaaagat ggcgaacctt gccaccacgt aaaaaattaa 48300

cgaaatcgaa taccgtttaa agcgcctgta agcgattcta gcgggcgcaa atttcattaa 48360

ggtgattcta tgtctgagtt ctcaaaagtc gaagatatgc cgattggcgc aacggtaact 48420

ggtatccaga tgagtgagtc ggtcgatacg attactccgc tggccttccc ggttacacag 48480

gtcgaaaccg atagcaaaaa aggtttcatc ttcatctaca aaaatttcaa cgcccctctg 48540

cgtgtcgaga tcttcattgc tcgcggaacc tgggtagaat gggagaaggc ttaa 48594

Claims (10)

1. A drug-resistant Klebsiella pneumoniae RDP-KP-20004 is characterized in that the phage has a polyhedral three-dimensional symmetrical head, is wrapped with nucleic acid, has a diameter of about 70nm, has a tail with a length of about 170nm, has a tail sheath, has a neck connected with the head and the tail, is a coccales Myoviridae, and has a preservation number of CGMCC No. 21407.

2. The Klebsiella pneumoniae RDP-KP-20004 of claim 1, wherein the phage maintains good activity at pH 5-9 with a potency of 1 x 10⁶Above pfu/mL, at pH 7, the potency is highest, suitable for birthGrowing in a neutral environment; at pH 12 and pH 3, the phage still survived, which was acid-base tolerant over a wide range.

3. The Klebsiella pneumoniae RDP-KP-20004 of claim 1, wherein the phage has a stable titer at a temperature range of 35-55 ℃ and is maintained at 10¹⁰More than pfu/mL, still has biological activity after being acted for 30min at the high temperature of 80 ℃, and has better tolerance to temperature.

4. The Klebsiella pneumoniae RDP-KP-20004 of claim 1, wherein the RDP-KP-20004 is capable of infecting Klebsiella pneumoniae with high efficiency and the optimal multiplicity of infection is 0.1.

5. Use of the klebsiella pneumoniae RDP-KP-20004 according to claim 1 in the preparation of a medicament for preventing and or treating diseases caused by klebsiella pneumoniae.

6. Use of the klebsiella pneumoniae RDP-KP-20004 according to claim 1 in the preparation of a feed for preventing and or treating diseases caused by klebsiella pneumoniae.

7. A bactericidal composition for the control of klebsiella pneumoniae comprising an effective amount of the klebsiella pneumoniae RDP-KP-20004 of claim 1.

8. The bactericidal composition for controlling klebsiella pneumoniae according to claim 7, further comprising an antibiotic for controlling klebsiella pneumoniae.

9. An additive for preventing and/or treating diseases caused by Klebsiella pneumoniae, which comprises the Klebsiella pneumoniae RDP-KP-20004 defined in claim 1 as an active ingredient.

10. A cleaning or disinfecting agent comprising klebsiella pneumoniae RDP-KP-20004 according to claim 1.

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