Bacillus cereus phage dz1 and application thereof

Abstract

The application discloses a bacillus cereus bacteriophage DZ1 which is not easy to produce resistance and an application thereof. The bacillus cereus bacteriophage DZ1 has a preservation number of GDMCC No: 63387-B1. The bacteriophage DZ1 can specifically lyse the bacillus cereus, and is not easy to make the host produce bacteriophage resistance in the use process, avoids the phenomenon of tolerance to the bacteriophage due to strain mutation in the subsequent process, and can better remove the bacillus cereus. Meanwhile, the bacteriophage DZ1 has good tolerance to high temperature and acid and alkali; in the removal of the bacillus cereus with rice matrix and milk as food objects, the bacteriophage DZ1 shows good bacteriostatic effect. The bacteriophage DZ1 and a gene product coded thereby can be used for preventing and controlling the pollution of the bacillus cereus in food, clinic and environment.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a Bacillus cereus phage DZ1 that is not prone to developing resistance and its applications. Background Technology

[0002] Bacillus cereus is a Gram-positive bacterium that can cause foodborne illnesses. It is widely distributed in nature and food, and its ability to produce persistent spores makes it highly susceptible to food contamination. Bacillus cereus produces a series of virulence factors that, when ingested, can cause gastrointestinal infections, leading to abdominal pain, diarrhea, and in severe cases, death. The *China Health Statistics Yearbook* indicates that Bacillus cereus, along with Salmonella, Vibrio parahaemolyticus, Staphylococcus aureus, and Escherichia coli, are major microorganisms causing foodborne microbial contamination in my country. Furthermore, existing research has found that foodborne Bacillus cereus can develop resistance to antibiotics such as erythromycin and tetracycline. Therefore, using antibiotics to control Bacillus cereus is extremely challenging, and the development of new control methods is urgently needed.

[0003] Bacteriophages are one of the effective tools for addressing bacterial antibiotic resistance. A bacteriophage is a virus capable of infecting bacteria, fungi, and other microorganisms. It affects the normal metabolic processes of the host bacteria, multiplies and amplifies within the host using its genetic material, ultimately leading to bacterial lysis and death. Therefore, the use of bacteriophages and their derivatives has broad application prospects in the control of pathogenic microorganisms. In livestock and poultry farming, there are already bacteriophage products for preventing and treating enteritis caused by *Clostridium perfringens* in breeding rabbits and for purifying the *Clostridium perfringens* environment. Clinically, bacteriophage perfusion and flushing therapy are also used to treat urinary tract infections caused by pan-drug-resistant *Klebsiella pneumoniae*. Compared with other physical and chemical sterilization methods, the use of bacteriophages in food has advantages such as high specificity, safety, reliability, and no chemical residues. Currently, many bacteriophage preparations have been approved for use in various fields internationally; for example, the U.S. Food and Drug Administration approved List Shield in 2006. TM The formulation is used as a food additive to control Listeria contamination in meat products; the E. coli O157:H7 targeted phage formulation has also been approved for use by the USDA organic certification for the prevention and purification of E. coli O157:H7 in livestock farming environments.

[0004] Meanwhile, the application of bacteriophages also faces challenges such as insufficient bacteriophage resources, a limited number of phages capable of sustainably inhibiting host bacterial growth for extended periods, and a low likelihood of inducing phage tolerance in the host. Furthermore, bacterial mutations caused by bacteriophages can lead to phage resistance, ultimately resulting in secondary contamination of samples. Summary of the Invention

[0005] The first objective of this invention is to provide a strain of Bacillus cereus bacteriophage DZ1, with accession number GDMCC No: 63387-B1.

[0006] A second objective of this invention is to provide the application of the above-mentioned Bacillus cereus phage DZ1 in the preparation of antibacterial agents.

[0007] Preferably, the antibacterial activity is the inhibition of Bacillus cereus in individuals, food, or the environment.

[0008] Preferably, the antibacterial activity is to inhibit Bacillus cereus in food, and the food is rice or milk.

[0009] A third objective of this invention is to provide an antibacterial agent containing the aforementioned Bacillus cereus phage DZ1 as an active ingredient.

[0010] The fourth objective of this invention is to provide primers for detecting the above-mentioned Bacillus cereus phage DZ1, comprising: forward primer sequence: 5'-AGTCAGCATGTTACAAGCCA-3'; reverse primer sequence: 5'-CGAAATGGTGAAAGCGAAAATGG-3'.

[0011] The fifth objective of this invention is to provide a method for detecting the above-mentioned Bacillus cereus phage DZ1, which involves detection using the molecular target gp6 of the above-mentioned Bacillus cereus phage DZ1 or detection using the above-mentioned primers.

[0012] Preferably, the nucleotide sequence of the molecular target gp6 is shown in SEQ ID NO.1.

[0013] Preferably, the detection using the primers described above includes the following steps: extracting DNA from the sample, and then performing PCR amplification using the primers described above. If a band size of 208 bp is obtained, then it contains Bacillus cereus phage DZ1; otherwise, it does not contain it.

[0014] The sixth objective of this invention is to provide a qPCR detection method for quantifying the above-mentioned Bacillus cereus phage DZ1, which includes the following steps:

[0015] a. Use the primers described above to plot the standard curve for Bacillus cereus phage DZ1;

[0016] b. Use the primers described above to perform qPCR amplification on the sample, obtain the ct value of the sample, and calculate the content of Bacillus cereus phage DZ1 in the sample according to the standard curve.

[0017] Advantages of this invention:

[0018] The bacteriophage DZ1 of this invention has a short latency period of 10 minutes.

[0019] The phage DZ1 of this invention exhibits good antibacterial effects and is not prone to inducing phage tolerance. It effectively inhibits the growth of host bacteria when the MOI is between 0.01 and 1000, and no resistant strains are developed within 18 hours.

[0020] The bacteriophage DZ1 of the present invention has good thermal stability and pH stability, and can remain stable at temperatures between 4 and 55°C and pH values ​​between 5 and 9.

[0021] The phage DZ1 of the present invention has a relatively wide host range and can lyse 21 different ST types of Bacillus cereus, including the vomitogenic Bacillus cereus ST26.

[0022] The bacteriophage DZ1 of this invention has a good antibacterial effect against Bacillus cereus in rice matrix and milk, and is not prone to producing resistant strains.

[0023] The virulent bacteriophage DZ1 provided by this invention can specifically lyse Bacillus cereus and is unlikely to induce phage resistance in the host during use, avoiding subsequent phage tolerance due to strain mutation. This allows for better eradication of Bacillus cereus. Phages that are unlikely to induce phage resistance in the host can inhibit bacterial reproduction for a long time, effectively suppressing the host's growth and providing better protection against Bacillus cereus contamination. It has good potential in practical product applications.

[0024] In addition, phage DZ1 is resistant to high temperature and acid / alkali conditions, has strong environmental adaptability, is not easily inactivated during application, and has a good inhibitory effect on target bacteria in rice substrate and milk.

[0025] In conclusion, bacteriophage DZ1 shows promise as an antibacterial agent for controlling Bacillus cereus contamination in food. Bacteriophage DZ1 and its encoded gene product can be used to control Bacillus cereus contamination in food, clinical settings, and the environment.

[0026] Bacillus cereus bacteriophage DZ1 was deposited on April 23, 2023, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong Province, 510070, China. The accession number is GDMCC No. 63387-B1. Attached Figure Description

[0027] Figure 1This is a schematic diagram of a plate containing phage DZ1 lysing the host.

[0028] Figure 2 This is a transmission electron microscope image of bacteriophage DZ1.

[0029] Figure 3 The figures show the results of validating the characteristic sequence of phage DZ1 and detecting phage DZ1 using a high-resolution melting method. A is an agarose gel electrophoresis image comparing the characteristic sequence of DZ1 with other known phages; B is the melting curve; C is the normalized melting peak diagram; and D is the standard curve between the ct value and the DNA concentration of phage DZ1.

[0030] Figure 4 This is a schematic diagram of the one-step growth curve of bacteriophage DZ1.

[0031] Figure 5 This is a schematic diagram of the killing curve of bacteriophage DZ1.

[0032] Figure 6 This is a schematic diagram illustrating the thermal stability of bacteriophage DZ1.

[0033] Figure 7 This is a schematic diagram illustrating the acid-base stability of bacteriophage DZ1.

[0034] Figure 8 This is a schematic diagram illustrating the effect of phage DZ1 on inhibiting the growth of Bacillus cereus in rice substrate (A) and milk (B). Detailed Implementation

[0035] To further illustrate the technical solutions and implementation effects of the present invention, the present invention will be further described below in conjunction with embodiments and accompanying drawings.

[0036] Example 1: Isolation, purification and proliferation of bacteriophage DZ1

[0037] 1. Sample source and processing

[0038] The samples used for phage isolation were sourced from surface water in Guangzhou, Guangdong Province. The collected water samples were filtered through gauze and then centrifuged at 10000×g for 20 min to separate impurities. The supernatant was filtered through a 0.45 μm filter membrane to remove some environmental bacteria. Magnesium sulfate was added to a final concentration of 50 mM, and the mixture was allowed to stand for 30 min to enrich the phages. The phages were then filtered through a 0.22 μm filter membrane to adsorb onto the membrane. The filter membrane was then sonicated for 5 min in a phage buffer solution (3% beef extract (w / v), 3% Tween 80 (w / v), 50 mM NaCl, with water as the solvent). The treated sample was then filtered through a 0.22 μm filter membrane to remove bacteria, and the filtrate was stored at 4°C for later use.

[0039] 2. Isolation and purification of bacteriophages

[0040] Add 30 μL of the initial Bacillus cereus 2432-3 culture medium (containing 1 mM CaCl2) and 100 μL of the preservation filtrate to 3 mL TSB medium (containing 1 mM CaCl2). Incubate overnight on a shaker at 37 °C and 200 rpm. Then centrifuge the bacterial culture at 12000 × g for 1 minute. Filter the supernatant through a 0.22 μm filter membrane to obtain the unpurified phage filtrate, which is stored at 4 °C.

[0041] Unpurified bacteriophages were purified using a double-layer agar plate culture method. 100 μL each of *Bacillus cereus* 2432-3 cultured to the logarithmic growth phase and the unpurified bacteriophage filtrate were added to 4 mL of TSB top agar (containing 0.4% agar and 1 mM CaCl2), mixed thoroughly, and poured into a TSB solid plate (containing 1.5% agar and 1 mM CaCl2). After the top agar solidified, the plate was incubated upside down at 37°C. Once plaques appeared, a single plaque was picked up with an inoculation needle and streaked onto the TSB solid plate. Then, 30 μL of a mixture of overnight activated host *Bacillus cereus* 2432-3 and 4 mL of TSB top agar was poured onto the plate while still hot. After solidification, the plate was incubated upside down at 37°C. This process was repeated 4–5 times until uniformly sized and shaped plaques appeared. Figure 1 This means that purified phage DZ1 was obtained.

[0042] 3. Proliferation of bacteriophage DZ1

[0043] The purified phage DZ1 was propagated. The host bacillus cereus 2432-3 was cultured overnight. 500 μL of the bacterial culture was inoculated into 50 mL of 50 mL TSB medium containing 1 mM CaCl2. After 1 hour of incubation, the phage plaques were picked and co-cultured with the host bacteria. After further incubation for 5–6 hours, the culture was centrifuged at 10000 × g for 10 min. The centrifuged product was then filtered through a 0.22 μm filter to obtain a titer of 1.02 × 10⁻⁶. 9 PFU / mL of phage DZ1.

[0044] Example 2: Morphological observation of bacteriophage DZ1

[0045] Take 10 μL of high-titer DZ1 bacteriophage after proliferation and culture, place it on a copper film with a carbon-plated mesh, let it stand naturally for 15 min, and then absorb the excess liquid with clean filter paper. After drying, stain with 2% tungsten phosphate for 10 min, and after air drying, observe the prepared stained slide under a transmission electron microscope. Figure 2 It was observed that phage DZ1 is a long-tailed phage. The phage DZ1 of this invention belongs to the order Andromedavirus.

[0046] Example 3: Phage DZ1 Genome Analysis

[0047] The genome of phage DZ1 was extracted and analyzed. The DNA of the high-titer phage obtained in Example 1 was extracted using the phenol-chloroform method, and a genomic library was constructed. The phage DNA was then sequenced using Illumina MiSeq. Analysis revealed that phage DZ1 has the following nucleotide sequence:

[0048] TGTTTAGCTGCTTCAATTTCTCGCTTAGTTTCTTCACTCATAGCTGGGAATCCAAGCTCCTCACGTAACCAGTCCTCA

[0049] GTAGGTGCAATTACTCCAGCAGTAATCATTTGAGTGAATACAGTAGCTAGTTTAGTAATGTCCTCGTTAGCTAGTGGCTTG

[0050] AATTGAATACTTGGGTACTCTTTAGCGTTTGGAAAGTTGTAATCTATTAGAGGGCGTACTACTTCCTCCTCAAGCAATGCT

[0051] TTAAGGTCACGCTGTAGAGCCTCTAAACGAATCATGAAAATGTCAAATTGGTTACCTGAGAGCGCATAACTACCAGATTG

[0052] ACCACGAGAAAGACCAAGCAACATAGGTGGTACTAGCATTGACTCCATAATTTTGCGGTCATGGTGCTCAATATAGCCAA

[0053] TAAAATCGACAGGTTGCATCTGAACAGCCTCCACTTTATCGCCACCACTAATAGCTAAACCAGTCATACCGTTAATCTTG

[0054] CTCAGCAATCTTTTCATTTTCCAACGTCGTTAGCATCTGTGGTTGTACCCACTAGCAGCGGTGTGCCATAACGCTCATAA

[0055] GCAATGTTAGCAAAGCGGTACAGTTTGTCTTTAATGAGCCAGTGCTTGTAAGTCGTTCTCAGAGCTGAATTACCGTATAG

[0056] GTTACCAAAGCGCTTGTCATAAGTGTAGAGAATAACCTTGTTAGCTTGAAGACGTATTTCTTGTGAGCCAATGTTCTGCT

[0057] GCACATACTCAAGGTCACCAAACTTGTCTATTTCAGTTTTAACTTATGAGGGTCAAGTACTTTTAGCTTTTTTAAAACAA

[0058] TTTGTCCTTTGGGGTTGTACTCAAAGACTTTTTCTGTTACGCTATAACCATATTCTAAAGCGGAAAGCATTTCTGCAATGA

[0059] CATCCTCCATATTACCACGAATAGACTCAAAGTTTTGTAGTACAAACTCTGCATACTTCCTCGATTCCTCCGAGTTACCTG

[0060] TTACAGTGAACCCTCTCGCTGTACTTGAAAGTTTAATCATGTCTAATGCAGCTTTTACTTGTCCGTCAGTCATCATTTTGT

[0061] CGTAAACATCTAGGTCAAAGTCACTCGGATTGTAGTCTAATTGGTCTGGTCGCTCGTTAGATGTATCCTTGTAGATACCAA

[0062] CCTCGATAGCCAATTTCGCTAAGTCTTGAGCTAGTTTCTGCTCAGCGTACACATTCCAAGGCAATAAGTGTTTAAATTCGT

[0063] TATATAATCCCATAGTTTTCCTCCTTGGCTTTCTGCTCCTCTAGGGAATCCAGCCTTATATTAAACTTATGGAGGGACTGTT

[0064] TACCAGTCTGGTAGGTCGTCCCCATAGTCAAGAGCTTGCTCTACTCTGTCAATATCTCTTTCTACGTAATCCCACTCAAAG

[0065] CGATTCATAACCTCGTGTAGTGCCTCCCTCACATAGTTTAGAGCGTGGAAAGCATCGTCTGGAGTTCTGTGGTCGTAGCG

[0066] TTTAGCACCTGTAGAGGTAGTACTCTCAGTGAATATCATTTCTATAGCTGTCCAGTGGTCAAAGAAGTACTCAACGTCTT

[0067] CTGGTTTCTTGTAAGGGATAATCAGTTGACCTTTTTGGAACATGTTAATAAGCTTATCCATTGAGTAAGTACGGTCAACCT

[0068] GTAGGGTTGAATCGTTATAGCCTTTGTATTCACGCTTTCTAGGGTCTGCTGAGTACGTCACGTAGCGACAACTAATAGCAT

[0069] TCCTTCCATAGATACCATAGAGCTTTTGCGACTCGTAAGAACCATAACCGATGTCACCAACTATTTTCTCTACATTGAAGC

[0070] GACCATATAGGTAGCAAATGTACTCAATGAGCTGGTCATGAGTTGACTTTTCGCTCTCTGGGTCTGGTAGCCAGCTTTCTA

[0071] TATGGTCAATAGCTAGTTTGTCTGCCTCCATATGTCCAATGCAAATAATCGTTTTAGACTTGCCGCCACTACCGTAGTCAA

[0072] TCCCCATGCAAGTTGGCGTATCAGAGTATTTCTTGAGAGACTGCTTCTTGTCAGTACAAGCTAGAACATCCTCCAAAGTA

[0073] ATTGGCTGCTCGTCACCGCTGTAGAACTCTCCCAGTACCTCATTAGCAAACGTCATAGCGTCCATTGTTTGGAAGTCACG

[0074] CCATATCTGGTTGGCACTAATCCAGACCATGTTGAGCTGGTTGAATAGGTAGCCACTGTACATTTTGTTCATAGGTCTTGT

[0075] ACGAATCCACTGACCATTTTCACGGTTTAGCTCCTCGTCACACTTCATACAGCCAAAATAGCGACGCTCAGTTTCTTCTC

[0076] CCTCGTTCAGTACCTTGATGTTATTCATACTCATAAATTGGTCATGACCGCACTTCTCACAGCATACTGACCACTTTTTCT

[0077] GGTCCGACTGACTCCAAAGTACTCTATCGTAGTAAGAACCTTTTTGTTTTGGCGTACCAGTAAAGAAACATCTACCATTT

[0078] AGCTCCGTTACTGGGTCAGTTATCTCACTATGAGAAACAGACTTCTCGATTGACTCAATAGCTGTTTGCGTAATGTCCTG

[0079] CACCTCATCAAAAAATACAATATCTCCAGAGATACCTCGCAAAGCATCCCCATCAGCCCACGCTGAACCAAAATAGTATA

[0080] CTGTTGAGTTCTTCAAGGCAATAGCTGTTTTAGCATCCCTCTTAGGGTCAACCATTCCTTCAAGTATTCCCTGTTTGGATT

[0081] CTGCAATTGATTTACGGAATCTATCGTTAACAAACCTCGTAGTCTGCTCTTGTCGTGGAGCTGTATAAGTAATTGTTGTGT

[0082] GAGCTCTCTGGAAACCAGCGAACTTAGCGAGGCGGTCAACTGTCTCGGATTTCTCTACTTGTCGTCCAGCGACAATAAC

[0083] GATACGAGGATGCTGGTCTCTGTAGACATCATGCAAGTGACCTCTGTGGTCAAAGCGAAAGGGTTTCCCCTTAACAGTT

[0084] CCAGTCGTCTCAGTGAAGCCTACTGGGTCTTTCATTACGTTTCTCATGTGCTTGAGCTGTTCAGCTGTTAACTCCATAGC

[0085] GCACCTCCTACAAAATTTGCTTTATAGTGTAAAACTTTTCATAAAAAAAATGAGGTAGATAATACTAAAGGGGTGGGGACT

[0086] GTCAGTTGGACTAGGGAGGCTACAAATTTGCTAAAGTACTCCCAGTTCATCTATAATAATCCACTATGCGTGGTGGACAAG

[0087] GGGGGTACACCTCATAATTATTCAAGGAAGTTGCATAACCATACAGCCAACTTTGCATAAAAGGCGAATAAGCAAGTAA

[0088] AACCGCATGGTTAAGCGATTCCTTAGTTTACATAATTAAGGTTATAGGAAGTTGTTAGTTATCGTCTCCAAAGTCAAACAA

[0089] TTGGTTTAACACAACAGTATTATCTCCTTTGTTGTTCTTATTCTCCTTGCTCATATTAAACTTACGGTCAAAGCCAAGAGC

[0090] TTGCATGTAGCGTAAGAACTTTGTATCATAATCATTGTAGCTATTACTCTCATCTACTTGCTGACCAGCAGCAGCTATCTCT

[0091] TTACGTTGGTTGAGTATGAAGTTTCTTAGTGCTCTGTCTAGCAGCATCAGGTTAGCTAAGTCTAACTCATACGCTTCCTCA

[0092] TATGCATTCATCATGTACTCATAGAAGGCTTGCTCCTCCACAGTAAATACAAACTTAGCATACATACCATGTATCATAGCAG

[0093] CTAGTGGTGACAGGTTAGCTAGGGCTCTCTGCTTACCCTCCTCTGTAGTAGGACCTTTACAGTACCCACCATGAGCAGCA

[0094] CACCTAGCAGCACCCTCCACAGGTGGCTTAGTGCATATCTTACCAGTGGCATTGATAGCACCGCATATAGTAGTGGCTTC

[0095] ACCAAGGTGCTTCCTCACAAGCTCAGCCTTGCTCTTTCTGGACTTCTCATACTGCATGTCATTAATCTCATTAGCTACTGC

[0096] CTCAGCATCCTTCTTGAGTCTGCCAATTAGCGCTTTCTCGTGGTTATTCATTGCGTTCCCTCCTACAAATTTCTATTCGGTT

[0097] TTACCAGTCGGACACTTCTGGTGCGTCGAGGGAAAGGTAAGCTATTTAGCAACCCCTCTAGTCCCTCCCTCAACCATTGA

[0098] TACGATTGAGTTTTCTAACTCTCAGCTTTTACTCTCTCGTGTTCCTCTCTAGCAATTTCCAACAAAGTTTTTATACAAATTT

[0099] GACCACTCAAAACTAGTTCTCTCTCGTCCTTCTCATAAAGCATAAAACCATACTCATTGAGTTGATACTTCCTAAATAACT

[0100] TAACTCGTTTCAAGCTCCTTTGTCCATATAGCCTTTGACTTCATAAATACGTTCCATTGAGGTAGAAAAAAAGTCAGGTA

[0101] AGTAACTTTCTCGCTCTGCTCTGTAGTAAAACCTCTCTGGCTCATATACAAAGTCAATACCTAGTGAGTGTAGGTGTTCA

[0102] GCTAGTTCTATCTCCCAGTTGGACCTAAAGAGCATATCTAACTTGTGACTCTTGAACTTCCTACCATCAGTCCACTTTTTA

[0103] GCTTGTTTCTTTTTAGGTTTCCCTCCCTCATGAATATCTCTTAACATATAGTTACACCTCTGTAGTTTTGACTAAGTAGTAC

[0104] CAAGAGTTACTATCTCTCTAGTATTCTATTTGTCTAACTCATAAGTTCTTTTGTTTTATCTTTTTATGAGCTAAACCTAATT

[0105] AGTAATATCTATTTAGTTAACTATTAGTTAGTAACTCCTCATAGTACTTATGAGAACCGTGTTTACAAGCAACGCAAAAAA

[0106] GCCACCTTTTAGGGTGACTTCTGAGCATCTTGAGAGCTTATTTTAACATGTTGGAACTTCCAAACGTCATCTTGGGGTCG

[0107] ATTTACTACATGTTAGTGCTTTTTTAGTTGCATTTATAGTGCTGGTGTTTACTTTCTGGCACTTCAAAGTGTTGTTTGACTTCC

[0108] AGTAGGACCATAAGCTTGCAGTTTAATAGCTCCAGCTCCTTGTAGGCTGCAATCATGTCATTCTTCAAGAATAATACGTCA

[0109] AAGATGTCAAATGAGTGCTTACAACTCAATCTGGAGAGCTTTCTCTGTATTTTCTCAAGCAATGCAGCTTTTCGCTCCCT

[0110] CAGGTCGTCAATTTCCTCGTTGATATAGTTCATAGTGTTTCTCCTCCTCTGTTTTGTACTACAGAGGTCATTCTACAGTAA

[0111] AGAAGGGAAGCTGTCAAGCTCCCCAAGCTATACTCAGTACGTAACCGACTATCCAGATATAAACCAAGCACGCTGCGAC

[0112] CTTCAAACTACCTCTCATGCTCATTTCTTCTCCTCCAGTCCTGCTAAAAAGCAAATACACGTTGCTACAACCATTCCAAC

[0113] CATACCACCCAAGCCAAACACCGCCAAGTATACGTCCCAGCTCATAGTAGCTCCTCCAGTTCTTCCTCAAGAGATTTAAT

[0114] GACTCTTTTGCTCTCAGCTATCATCTTATAGAGGCTTACAAGGTGCTTATCACTCTTACCATTACCGATAGCTTGCCATAGT

[0115] CGCTGTTGGTACTCTCTCAGGTCAGCAACCTCCTCAGCATATTGTATAATAGTTGGTCGAGCCTCAGCGTCCCTCTCACA

[0116] AGTACAAGACCACTCACAATTGCATCCCATCATTTGCTCTCCTCCTCATGGTACATGTCAATTAGCATTTGTTTTATCTCAC

[0117] TAGCTGTCAACGTCTCACGGAAAGACAATACTGAGCTCTGACCTCGTTGCATTTTCTCAAAAACCTTAATCATAGCCGCC

[0118] AGTTTATACTTGTCAAGCTTCTTCATTAGTTTGCACCTCCTTGCAATGCTGTTAGCTCGTTTTGAGCCTCTATATAGATTTT

[0119] ACCAAGATGTCTAATTGAGTACTCGTCAACTGACCAGTCACCTCTGGAGATTTTCAATACAGCCAGCATACCGCTGTCAA

[0120] GGTCATGAAGGTTATTCAAGATGTGCGAACTATTGGCAATTACCCACTCTTTAAACAACACGAGGAAGCCACGACGAGT

[0121] CTCATTTACAATAGAGCCAACTACAAGGTCAACCTCCACTGGCTCATCAGGAGTGTTGAACTCATCCACCTCAGCCAAT

[0122] CTGAATAGGTCTTGCTCTGCTTCTATCTTTAATTTAGCCAATTGACTAAGAGTGAACTCACCAACTCCATACTTGCCGCCA

[0123] ACTATTTGTACTACACGTTTTACGCTCTCTGGGAAAAGGTCAGTACCACTATAATGAGCGATTTTAGCATTATTAGTTAAA

[0124] ACGAACTTACGGAACGAAATGTCAAAGCCTCTGTAAGCATTCTCAGCAGCCTCTTGTATCTTGTGAGTTGGAACATAGTT

[0125] GTCACCGTCTTCATAGCTTACAACTTCCTCGTTAATCGCATCAATTTGCAACTGAATGCCAACTTTATGAGCTTTGACTTG

[0126] GTTGTTGTTAGCTTTGCGAGCTTTTTCTGCTTGGTAGTTGTGAGCCTCTTGAGCTGAAAAGAACACTGTACGCTCCTCTT

[0127] GAGCTGGCTCATCAGCTTTGTAGAATGCAATGAACTCCTCCATTTCTTTTAGGTCAGCTACAGTAGCACGACTAGCAGCT

[0128] TTGTCATACTCACCAGCAACAAGTAGGTCATAGAAGTTTTCACTCTTTTGCAAGCCCCACTCGCTAACGGCGTTGATAAT

[0129] TACCTTTTTAGCTTCCACAGTACCAAGATTCATCATTGCAGCGATTTTCTCAACCATTTCTACTTTTTTCATTTAAAGCTCC

[0130] CCTTTCGGTCATACTCTGGTATGACTCGTTTGATTTGTTACTCTTAGTATATGCAAAGTGTTTACTTTTAGCAACAACTTTT

[0131] GAAAATAATCGTTCGACAAAGTTCGACACTAAAAAGACAAAAGAAAAACCTCCCTAGTGGGAGGCTTAGCGAGCGTAT

[0132] TCTACTGAGTCAAAGTCTTCTTTAGCTAACTTGTT AGTCAGCATGTTACAAGCCATTGAAGCATGTTTC CAGTTAGTGAA

[0133] CTTATGAGAAAGTTCAAAATCTGTTACATACTCACCGTCAGCGTCCCAAGCGTCGCCTTCAATGACAAA CTCTCCGTCAT

[0134] ATTCTTCAACTTTTAGGATAACTTTGCAGCCTTCAATATTTTCGAATTGATTGATAATTCTCATTTTCG CTTTCACCATTTCG

[0135] TTAGTTTTCATCATCTTAACCAGCTCCCTTTTATTTTGTTTGACTTCTTACTCTTATTATATGCGTATTACTCCAGAATTACC

[0136] ACACTTTTTTATATTTTCTTTTCGACAAATCTCGACAGATAAAAAAAAAAGAGACCTTATTGGTCTCCTATAAAGTCCTCT

[0137] AACATAGATATTGCTTGCTCTATATGCTCACGAGCGTCTGAGTTAACGTCTACGTGGTATTGACCACCTTCAAGTGCTCTT

[0138] AGCAGCGAGGCAGCTTCAACCTGTGAGAGCTCAATTACGAGTTGTTTAGTTATTTTCTTTCTCATTATAGCCCTCTCCTTT

[0139] GCTTCTCTGAGTAATACAATAGCAGCGTCATACCAGCAATCATACTCACAAGTACGACTATGAAGCCCCAGAAGTCTCCA

[0140] AACAAGTAAGATACTAGCTCCAAGTACCCTATAAATAAGCAAATCACAACCACAAGAGCAAATATAAAAGCAATTGGTT

[0141] TCATGACTGCAAAGAAACAGTCTTTTAATAGTAGCCAGTTCATTAGTCTAAATACCTCCCTAGATAATCCCACCAAGCAG

[0142] AATTATTGTTTTTAATCATAACGTGTCCGTCCGAATCAATAGTGACCAGTGAAGGGACTCCATTCTCCTCAGCTGTAAAC

[0143] AATTTACGTCCGTCCTCATAAGTCTGTACATGCACGACAAACATTAAGTCAAACGCCTCAGTCAATCTTTCTGCAACAGT

[0144] TGGCTTTCTCATAGTATATGACCACACTTTTCGATTAATTGTTGAATCAGCATAGCTCTGACCTCTTTAACACGCTGTATAG

[0145] CTTCCTCCTCGGTCTCATATCGTCCATAGCGAGTGCGCTTACCTTCAACATTGAAATAGACAACATACTTACCGTGGTGAA

[0146] ACTCATAGTTCTTTGCTTTGGTATTCAAGTTGTTTAGTGTCCTACTCACAATCCGTAAATTCTCAAAGCGATTATCACAAG

[0147] TATTTCCGTTTATGTGGTCAACGAGATAGCCCTCTGGAGCCTTCCCTCTTACCAGTTGGTGCAAGTATATTTCCTCTTTTG

[0148] CTGACCACCTCACAATGAAGTATGGAGCATTCTTTTTCATTCGCACATATATAGTCCTGCCATCAGCTACTAATAGATTGTA

[0149] TATAGCCTCGTCTATCCAAACCTTATGACCTTTTACGTCAATTTCCATTGTCCTCAATCTTGGTCACCTCCTCAAACGTAAA

[0150] TCTCATTATCTTATGGTTATCCATATCGCCACCAACAAGCTTAACCTGTACTGCTGGCATGTAGGGAGTCTTAATAAAAGC

[0151] TTTCTCGATATAGGTTATAACTCCCTCGTCCTCTCCAGTCGGATTATGTTCACTTTCGAGCGAAAAGCTTTTGACTTTTACT

[0152] GAGTCGCCAATCTTTAACATTATAATCGTCCTTTCCAGTTTTGCATAATGTCATACTTTTTCGTGTAATTAGCCATTACTACA

[0153] TTGTCATGTTGGTTAGTAACATGATTTATCCATATAGTCACAACTCGTGCGTCTCTGAGAGAAAACACAGCGCACACCTC

[0154] AAAGTTATACTTCTCTGTAGTAGTGCGACTGCGTAGCAATATGCGTGGGTCGCCTCCCTTTCCGTGGTCACCTACATGAA

[0155] ACTCGATAAGGTCATACTTTGTAAACAGTGCTTTGAAGTCTGTCTCACTGATTCCACGACCAGCAAGACGTTTCTTTGAG

[0156] TAATCCGTGAAAGACCACATAAAACTAGGTATATCTTGAAACTTGCCAAGGAGGAAGCTTTCTCCCCCAGCAGCCATTT

[0157] GACTAACATGTTTTTTATTTTACAAATAGTTTGCATAAGACTTACCTCCTTGGAAATATTAATTACTGTACAGTGTTTAAA

[0158] GCTTCCAAAAATGCTACTCGACAGCGTGCCAGCGCATAGTCCAGTTCTAGCAGTTTTGGCTCTGCATCAACTCTCATAGC

[0159] TACATATAGACTCCAATCGAGCCCTTGCTCTGGAGAAGTAAGCTGGTAGAGCATCGAGCATGTCGTCATAAGTCATGTACA

[0160] TTTGCTCTTTAGTGAAACCAGCCTCAGCGCACATATGGAAGGCACGCTCCCAAGCGTCAATCTCTGCCTCGTGTACTCC

[0161] AAAGTGTGCTAGTGCATCGTTATTGCTAGTGAAACCTTTATCAGCGTTCTGAGCAGCATGACCTAACTCATGAGCCAGTG

[0162] CTGTTAGGTGGTACAGATTATTTATTCTTGCATAGTTGCCTTCTAGTTCGATTGTCATTGGCTTTTCCTTGTGATTCCAAAA

[0163] GGCTGCATTGTCTTTGTGGCACTCTGGGCAAAGCTTCAAGGTAACACCTAGGTCATACATTACTTTTAGTAAAATACCTATT

[0164] GTAGCTTTTAAACTCAGTAGCTTTAATCTCAAGATAACTCATTCTTCCTCACTCCTTTGATATTTTGGAATAATGCTTTGAA

[0165] GCCACCAGCAGCTTTTTCTAATTGCTTAACTGCATTGTCAACATCAAACTCAATCTTAGGGAATTTCACTGGCTCAACAG

[0166] GTTTAATTTGTAAGTCCTTTAACTCTTTGATTACTCCTCCGTCATTCAAGTGAAAGCTTACAGGAGGTCGAATATCTTGGA

[0167] ATAATGCAGCATTGGCTACTGGCTTGAAGGCTTGGTGTTTAGCAGCTCCAATATGTCTTATGCTTCCTCTGTGGTGTCCCC

[0168] AGTCGATAGCTACTGGTTTGAATGGTACAGCTTTTGGGAGTTCAACTGGTTGGCAGATTATCTCTAGAAAGTCAACAAGT

[0169] TTCGCATCTTCCTTGATGTTACTCAGGTCATACCCCTCAGCATCAAGATGAGCAGTCAAACTGTAGCGCTTGACAATCTC

[0170] CTCAGCCTCTTTCTCGATTGGCTTTTGAAACTCCTCCCAAGTACATAAGTTAGGTGCAACATATTGGTTAATTTCTGAGAA

[0171] GTCGTTTGGAAGTAGCTCTTTAGCTATTTTCTTAACTTGACGAGTCATTTCTTTTATCTCCTCATCAAAACCCCACTTGCTT

[0172] CTTACTCCGTTGGTGTGTACTCCATGCTTTTCGAATAGTCCATAATCTACAATGACTATCTTTCCCTCTCTAGTCAGCATCA

[0173] AGTTTTCTCCGTGTAGGTCCTTTGGATTCCAGCCACGCTTACAGCTTTCTCGGACAAACTCGTTAATCTGTTGTATCCAGT

[0174] GTTTCCAGTCAAACAATTGAGGTACTATTGAGAGTCCCATCTTTGCTGGGAAAGCGTTCATATTGTACATTACCTCTCCA

[0175] GTTTGCATACCGTAAAGTCTCTCAACTACCATGTATTGCTTACATTCTGAGTAAGCGTATAACTTTGGAGTCATTGGTAAG

[0176] CCTTGCAGAGCCTCCAGACACTCACCATCGACAAAGCGGTCCATGTCTATATCAGCTCTGTTGTATGTTTCATTTACCTTG

[0177] AGTACGTGGTCTGGACTGATAGCCCAAGTGCCTCCACAGTGACCTCTGCCAAGTCTTTCAAACATGTATTGCGGTATAGG

[0178] TATTTCTTGCTCGTTGATACTGATAATTTGCTCCATTTTTGCCAGAGCCTCGACAGAAATTTCATTAACTAATTTGAATTTT

[0179] GCACCTGCAATAGTAAACATATTAGCTTGCTCCTGTACAATCGTTAAGTTATCCATTGTGTTGCCTCCTTTGGTTTTACCTC

[0180] AGTCATACCTTGAGGTGTTGTCTTGCTCTGTACTTTATATTATGCAGGGTGTTTACTAAAGTTGCAACATTTTTTTTAATTA

[0181] TTTTAACTTATTCGTTCGACATTTCTCGACAAACAAAAAGAAACCCTTGCAGGGCAAGGGCTCAGAGGTTTTTCAAATAT

[0182] TCTTTTGGGTCTGCCATATAGTCCTCATAAGCTGATTCCCACTCATGAGAAAACATTTGGTTGCCTTGCTCCATGTCACGC

[0183] TGTATAAGTCGTTTCACGTATGTTGAAAAGTATGACTGCTTCATTGAGAAATGGAACAGCTTGAACTCCATAGGGTCTGC

[0184] TAATTGAAACGCCACTGATTTAACTTTTTTCATTGTTGACTACCTCCCAAGTTGTATTTACTTATAGTATGACCAAGCTCAT

[0185] ACCAATTATACTACATAGCTGAGCCAATTTCCTTGAGAACATTTAGGAAGAATGGAAGCATTTGTATGAGTACGTAACCT

[0186] AACCCAGCTCGCTGTATCATTTCAAAGCCACGGTCTGCACGGTTAAACATGAACAGGAAGCACGCACCAGCCATGATTA

[0187] CAGAGGCAACTGGAAAGGCGAAAGCGACCAGTATTTCAACTACAGGGTCAAGCGAATGAGCTATCACCTCTAGTGTAT

[0188] GTTTGCTTACAAAATGTGTTACTCCCTCAGTCACACCACCACCAGCTACCTCAACAGCTTTCTGCTCACAGGCGAACGC

[0189] TGGAGCTGCTGCAAAGGCTAAACTTAATGGTAAGACAGAGCCACCAGTCAAAGCTGGAACAGCTATTGCACTCAGCGA

[0190] GCGTACTACAGCCTTTTGCTTTTTCTTACGAGCTGCTTTCTCTTTTGTTTTATACTCACCAGACATGAACTCACTGAACTT

[0191] GATAGTTGTTTTACGTTTAAACATTTTGACCACTCCCTCTATTTAATGTCCATAATCGTTAATACTTGATGAGACAGACCAA

[0192] ACTCCTCGCACATCTTAACCAGTTGCTTACGTCTTAACTCAGTAGTAGTAACCCAGATAATAGGAGGAAAGTAACCAAA

[0193] GTGTTTAGTAATAGCTCCTCGCTCGTTCATACCAGAGTATGCTTAGCTTGTTTCTATTCTCACTCATTTTCTGTTGGTTA

[0194] TCAACCTCTAAAATATGGTAATAGCCATTCATCTGAAACCAAGCGTCACAGACTTTACTCTCTTTCTTGTCTCCCAGTTTA

[0195] ATTTCAACTTTCCAGTCTTTAGGTTGACCTGCATAAATATAGAATTGATTACGCATGATAGTATGATAAGTGTATTGAGTGC

[0196] GTCTCATAGCCTTCTTAGCTTGCGTATAATCCTTTCCATTGCTATTGAGGTAGTAAACTGTGTCGTACCCCTCACGAAAGC

[0197] TCTGGAGGTATTCTGAGAGGTCTGAGAGTATGCGGTTAACGTTGCGTATCTTACCTAGCTTGTGAATAGCTCCAAGTTGG

[0198] TTCCTAGTCAGAAACCGTGTTTCTTCAAGGATAAAAGTATTAGCTCCTGCCTCACTGATAATGCTTTCTTCTTCATCTGC

[0199] GAGCACCACCCTTTTACTTATATTAGTAATATATGGAGAGCTGACTTGGCTAATTACCTCAGTATTAATAAAAAATGTCTGT

[0200] AGTATTTCTTGCTTGTCAGCTAACTGCACTATAGCTCTACCCTTCACAGCTGGAAGCTTTTCAGCTCCTCCCTCATCAAGT

[0201] ACTACTTGACTGGCTGTAGAGTTACGAACTCGAAAGCATATCTTAACGTCACAGTTTTGTTTGACTGCACGAGGGATAAC

[0202] GTCACCAGTTGGGTACTGTGTAGCCACTATCAGTCTGTACCCTAAACCAGCACCGAGCCGAGCTATTTGGGACATGTAA

[0203] CGTTGACATTCCTCTTTGAGTTTCTTATCAGCCTTGTCAGTTGCTTCTGCTGGGTTCAGTTCTCCAACCTCGTCAATGATA

[0204] ATAAAGTGTCTCTTGAGCAAGCCAGCGTCCTGTACCTTCTTAGCTTTGCCTTTGAGTTTGTTCTGTACTTTACGCATGCCA

[0205] TTGTAAGCAGCCTCCAGAGCCTTGAGAGCTTCTTCTGGCTCGTAAGCAATGCTTTGGGTTTGCTTCAAGTTCTCGTAGTC

[0206] ACAGAGCTCTACTCCTCCTTTTAAATCAATGAAGGTAAACTCTGTATTGTTTGGCTGAGAGTGTAACAATGAAATGATAA

[0207] TACTATTGAGTAAGTTACTCTTACCGTATCTTGTAGCACCTCCGAGCCCTACATGTGGAGTCTTCTCAAAGTCCAGATACA

[0208] CCTCTTTACTGTCTCGCTTGACTCCAAGTGGTACTTTCCAGTCTTTTGTATCTTCTGTGAGGTAGGGAATCTCTGTAGGAA

[0209] ATGGTTGGTCATATACCTTAACGTGTAACATTCCATCAAAGCTGAGCTCTACTTCCTTGTGTCCTCCTAGCTTCTCCTTTTT

[0210] AAGAAAACTTTTGAGATTGAACTTAGCAGCTCTGTTATTCAGTCCATCTTTTAGGATTCCTATTTTGTCCTCGTAGTCACT

[0211] AAATTGACGACCTTCTGGAATACGGTACACATATTGGACGCCCCAGTCATGTTGTTTCTTTCGGACCAGCTGAGCTGTGT

[0212] AGGTCTTGCCTCCGTGCTTCACGTTCAGACCACTTAAAGTAAATAACTTTTGGATTCTTTGCTGGTCGTTTGTTGCTCCGT

[0213] TTTGCTTCAAGTATGCATAGAGAGAAATCCCTCCCATAAGCATAGTGCTTGCTGCCTCGAAAATCATTTTGCTTTCCCTCC

[0214] TTAGCTGCGCCACAGTGTTATATTACTGTAAGTAATAGTTACGTCTATCGAGCATCCTCTAAAGTGAGGTTGAGGAGCTAC

[0215] ATGACTGCATCTGGTATATACTCGGTGTGACTTCCAGCTGAGCAATTTAATTGGTATGTTATAAGGTATACACTAAGCAGA

[0216] ATGTCTTAGAACTCATCAGGCATAAAAAAAAGTAGAGACTTTTTAGTCCCTACCTAGTCATTGCGTTTATATGTTGAGCAG

[0217] TCAATCTAGCATACTTCTTATTCATCTTCATGACATGCTGACCACCAAGAGTGTTAACTACCAGACACCAACCTCTCCAG

[0218] AATCTCCGCTCAGCAGCAGCAAAGTGAATTGTATCTTTGTGAATGACCATTCTCTTTAGTTGGTACTTGACATGAATTAA

[0219] GTCACCAGCTCGTGTTGCTTTACTACCTTTGAACTTTGGAGTATTCCTTTCTATTGAGTTAATGTCTAGCGGTTTCCACGG

[0220] TTTCATGATAATCAGCTCCCCTTCATATTTATACTTTCAGTATATGTCAAGTGTTAATTATTGTCAACACGCTTAATTGAGTC

[0221] TTCTGAGCTGAATCCTTGAGGATACCTTTTCCGTAACTTCTCAACGTTCAGTTGTCGTACCTCTTTAGGGTCAATCCCAA

[0222] GAGTATCCATCAAGCGAGTAAAGTACCAGTGAGTGTCACCAAGCTCTTTCTTGAGTTCGTCAATGTCTAGAGGGTGACC

[0223] GTGAAAGACAACTTTCTTAATAGTGTCAACTACCTCGCCAGACTCTCCAGCTAATCCCATACAGAGCATAGTTAGTTCTT

[0224] CCTCTACAGTTGCGAACTTACCAGCTGTGCGTTGAGTCTCGACTCTGTAGCCTTCCTCAGTAGCAGCTAGGTTCATTACCT

[0225] TCAATCTTGTCAACTGCTTGTTGCAGTAAAGCAATGAGCTCCTTAGCTTCTTGTACTGAGTAGTAATTGTTTTTGTTGTCT

[0226] GTCTCAATGTTTACTTCTGAGGTGTCTCCAACTTCTGAGTTACTTCCCTCATATACATAAACGCTAATTTCTTCCTCCCACT

[0227] CACCCATTGAGCCAATATGAATAAGTCTTCCCATTAGTTTGTTACCTCCAATTTCTGTAATAAGTTTTTAACTTGTTGTCCG

[0228] AATAACTTGGAATCTCCGTTATTAGTAATAACGTAATCAGCACGCAACACATCAACAGCTCTTTCAGTTGGCGCATTAAGT

[0229] ATTTTCTCATTAACTTCCTCACCAGCCTCTCTCATTCGTTCAGCTCTCACAGACTCATCAGCTTCAATCCTTACAAACACA

[0230] CAACCTAGTTGCTCGCACGCTTGGTGCTCATTGAGTTGGCGCACATCAGTAAAGATATAAGCTGGTACGTTCAATCCATA

[0231] TTGAGCCAGTGTGTTTTTGTGTAGCAATACATCTGCTATGGTTGGTCTAACAAAATACCTCTGAGTCAATCTCTCTCATGAA

[0232] ATTGTACTTCTGGTAATGAGCTATTGGCTTTGGTACGCTTGGTATCATTGGAAAGTCATTGTGAAACATGCGCTTCATAGT

[0233] GTCTCCATAAGCTACTCTGCGAACATGGTAGCCAAGCTCCTCAGCTATCTTGAAAAAAGTATCCTTGCCAGCTCTAGTAC

[0234] GAGCCAGCAAAGCTATGTTTGGTAGTCTTTTCATCTTATCGCTTCCCTTCCATCATTTGTATTTCTTCCTTAACTAAACGCT

[0235] TGAGTTTACTTTCTGACCAACCATTGTACTTGCACTCACTAGCTATCTTGCTGCGAATATCCTTACCATTGAGTTGCTTCCT

[0236] CGCTAAGTTACGAGCCACTTTTCCTATTGAGCGCACCATGAGCCAGACCTCCTAGAATCCAATTCTTTTAGTATCGTCCTC

[0237] CGTAATATACAGAATGCTACGAGTTGGAATGTACAGGACTTTCTAGTGTAGTAGTCAACAACTTTAATAGCTGCTTCTGC

[0238] CAGTGCTTCCTCTGGTCCAATTTCGTACTGCCACTCACGTTTAACACATTCGTATCTGTGGAGCTTAATATTGAAAAGTTT

[0239] ACCTTTCATGCTTAGCCAACTCCTCTTTTGTGTGGTAGTAAATACTTTCTTTTGTGTCATAGTGGAAAATCTCAATGTATGC

[0240] TTTTTCGAAAGCATTCTTGCCTACGTGTTGACTCATCTTACTTCTGAGTTGCTGAGCGTTCCAGCCCCAGTCTTCTGAGC

[0241] TGTCTACATGCATTACGACTCCATTGTGTTTACGAATAATCAAGAGCTTTATCATTTAGCACCTCCAAAGAATCCCTCTATC

[0242] CAAGGTTCTACCTTCACAACCTCCTCTTTCAGTTTCTCAGCTAAAACAGTTATTTCATGTTGTGCTCCATTACCAGCCATT

[0243] CTCTTTGAGTAGAAGTCAAGCAGCGCTCTCAGGTTCACTGACAGTGTTAAGTTACAAGTTGCAGCTTGAGGTAGTACTG

[0244] CTCTTGCATCCTCTTGTGGCACACCATAACTAATGAGTGTATCATAAGCACGTTGTAAATCTTCCATTACTGAGGTGAAGA

[0245] ACTTCTCAGCATATTCTTTAGCTTTCACCTTCTCAGGGACTACATAACCAAAGCTGCCATGTTTGCTGTCAGAACTTTGTT

[0246] TTACGTACCGTTGAGACTGGACCGAAAAGCCAAACCCTACACGGTGGCGAGTGAGTTGAGCCAGCAGCGCTCTTGAAA

[0247] CTCCCTCTACAGCAAAGGTGAATGAGATATGCTTCTAGCGTACTAGTGTGACCACTCTTGACAATCATTCTAATGAGACGG

[0248] TCAATCTCCTTGCCTTCACCCTCAGTAGCTCGTTGCTTCAAGTATTTGTCTCCCTCAGCTACAATAACCTCGCTTGGCTTG

[0249] TTAGGTGAGTAACAAGTACGAATAGCTGTAAGAGCTATCAGCTTTGCTTCCTCGTGCAACATGTCTTTCTCGTCATAGATT

[0250] CCATCAATATTACTAACGTGTTCAAAAAACTCTTGAGATAGTTGAGTGCGCCATTAAAGTAATTTTCATATTAGTAAAG

[0251] TTCCCCTCTCAGCTTTTCAATTTGTTGTCTTATATATTCTCTTGGCTCAGCTGCCTCTGTGTGCAATTCCAGTAAGTCTGTA

[0252] TCTAACTTGTCGAGCCTCGTAGCTATTTCTTCTAGTGAGTTGAGTAGCTGGTTAATACAACCTCGTTTGTTGAACTTCTCA

[0253] ACTGCTCCTTTAAGTTGGTCATATATGGAAGGTGTCTCGGTTACCTCTCCAGTATACTTGCCATTCTTGTCAAACCCAGTG

[0254] AAGAACTCACGAGCTTCACACAATTTACAGTTACATTTTTCCATTAGATTGTACCTCCTAGTTTTTACTTGGTAGCGAGAC

[0255] CAGCCAGAGTCAAGTCAGTAGCCAGCCTCACTGTAGGTACTACCATTGTATTACTCTAGTCATACTTTGGTGCAAGCTTT

[0256] TAGAAAATTCTTTCCAGTAAAGCTAACCAACACATAATGAAAGCGAATGTATTGAAAGTATCAAACCCTTTAAACTCACC

[0257] AAGAAAAACTTTTCGCAAACAGCCAAGTACTACCACAGTCATATACAGAAAAGCAATGACATACAATGTAGTAAACAAA

[0258] ATGTTAATCATTTCGTATCACTCCTATAAAGTCTTTTATAGCTATCGGTAAACCTACAGCGAGTGCGATAACTCCAAAGATA

[0259] CTAAGCGTCCAGCTCATCCAGTGTACCCTCTCCGTCGCACAGTGGGCAGACTTGAATTTCACCTTCTGACTCCATATCAC

[0260] AAATTACCTCGTCCTCATCATACTCTCCCTCAATAACTCCTATTACTTGGTGAGGTAGGAAAAGTGCTCCTGTAGAGTGA

[0261] CACGCTGGACAGTGAAACGCTTTGTTGCCAACTGGTACGTCTCTAGTATCGTTGCTTTGCTCCCACTGACCGCACTGGCT

[0262] TTCTGGTAAAACCTTTACTCGACGCTCTCCGTTTTTGTCTGTACCGAAAAGTGTCTCTGCCAGCAAACATCGTACCGTTT

[0263] CCCCTTTGAAATCTTTCTTGTAAGCTGAGCTGTGACGGCAACCTAAGCAATTTTTCTTCTCCATAACTTGACTCCTCCAAT

[0264] TCATTTCTTAGTTTTTTAAGAGCTTTCTTTAGAGTCACACGCACTGTTGCAGGGCTTATACCAATCTCGTTACCTACCTCT

[0265] GTAGAGTTCCACTTCTTGAAGACTGAGAGCTCAATAACTTGACGCTGGCGACTTGTTAGTTTATTGAGAGCTTGGTAGA

[0266] GTTCAAGATATTGAGCTGGGAGCTTTGGGTCTTCCTCGTCCTCCACTTCCATTGCTTCATGCTGGCTTTTAATCTTTTCGT

[0267] CAAACTCAATTTCGTTAATCTGAGTAGCCAAGTACTCCGAGAAAAAGCGGTCAAATACTCGCTGGTGAAGTTTAGCTCT

[0268] CACGTAGCGCTCAAAGCTTCCCCTCTCTGGGTGGTACTCGTTAACCAAGATAATAAATTCTAGTAAAGACTCTTGGAACA

[0269] AGTCCTCGTAATCTTGAAAACTAGGGTTTGCCCACAGCTTTTGTAACGCTTTAGCTTTGCGAGGTGGCTTAGTAGGTCTG

[0270] TGAAGTATCACGCTCAGTACATCTACATATTCTTGCATCAGTGACCACATAGCTTCACTGTCTCCATTCTGAGCTGCAAGT

[0271] ACCATTCCCTCATTGACATGTTTGTCGTGCTGCTGTGGAGCTACCCCAGATAGTGCTAAACTCATTAGCTTTTGCGTATCA

[0272] TTCATTGGTAAGTTACTCATATTATTGTTCCCCTTTTCGACTAGTTTTTTAACTCGATTGACTGTTACAGAGTACTTTGTT

[0273] GCTAGGTGTCCTATTGTATCTTTGGTGCTTACATTCCGAGTTAAAACTATAGAGTTAAAGTCTTTGAGTATAGCTTTCTCA

[0274] AGCTTCTCTTTTTCTTTCTTTCTAGCTGCTCTGGCTTTCTGTAAGGCTTCAAACTCTATCCAGCCACCATCTATTTTAGATT

[0275] CACAGATACATATGAGCTGGACATCAAACTGCTTGGCATAAAGCTTTTCTTGAGTTTAAACTCTGGAGTCTTCATACCTT

[0276] TAATGTCAATCACTTTTGTTGTTCCGTCGATGTACCAAACTTGGAAGTCAGCTTTATATTTAATAGCTGGGTAGTCCTTAC

[0277] CATGCTTTGGAAAGCTTCTTGTAGTACGAAAGCTGGCTGCATAGTAAACTCTTTAATTTCACCCATAGCCTTTCAATCTCA

[0278] TGAGGTGAGCATAGTATCTGCTCTCTACCTCACTATCAAACTTAATGCCATCTACGACAATTTTCTTGGCGTTAAACTTAC

[0279] TCATAACATCCAACCTTTCTTATTTATAATATTACTACAGTCATACCAGAGTGTCAAAAAAATTCGTCAAACTTTTTTCAAT

[0280] AAGGAGAAAATTCTCCAAAGTGCGCCTTGTGAGCTTGTTGGTACGCTTGTTGAGCTTCTTTCTCTGTATCAAAATGACCG

[0281] AGATATTGTTTGTTTAATTGAGCGTAATACTTCTCAAGTCGAGGACACCAGTAGTAACCCTTTGCTTTACGGTTTCCAGCA

[0282] TTCTGTGACTTATTAGCAACTCTCAAATTTTCTTTGCGGTTATCTAATTTGTTACCGTTTATATGGTCAACAATTACGTGCC

[0283] TCTCAGTGACTCCCATGAGCCAGCGGTGTAATAAAACAGGTCGTCTGTTTACGTTAATAGTTGCATATCCGTGGTTTCCA

[0284] ATTCTTATAGCCTTCTCATAAGCAAAATCATCATCAACAATTATTTGTCCACCAGTTACTTGTAGTATCAATTCACAATCGC

[0285] TCCCCATGAATCGCCTTCAAATGGTTTACATTCCTTATAGTGAAAGCAAAATTTACAACCAAAACCATCAGTGTACTTGTT

[0286] CCATGTATCTGTACGGACCGACTCACAAATATCTACAACTATCTGTCTAGCTTTTATCATTGCGTCAGTAACGTTAAACTC

[0287] ACAGACAGGAGCTCTCTGGTTCGTATACTTGTAGTGTCCTCCTCCTTGGTGTATAGCTTTCTGCCACTTAGAAATGACTG

[0288] GGTGGAAATACTGCACCGCTGCTGGGTAAGCTCCATGCTGCTTGTAAATAGCTTGAGCGTATATAGCGAGCTGTATATCA

[0289] GTGTTTAAAAACTGTTTAGACTTAACACTTGAGCCAGACTTATAGTCCGTTACAATCCAAGTTGAAGCGTCTGAGGGGT

[0290] CACCGTCTATGCGGTCAATATATCCAAGCACTGGCGGTATATCATCGACTAGTTTTGTCTCAAACTTTTTCTCTCTATGAAGG

[0291] TTGGCTTCTGGTCTTTGTAAAACTCATAGAAATTCCGTATAGCTTCACAACCTTTTTTGTACATCTTAATGTAGTCTTCCTT

[0292] GTCTTCAAAGTACTCTGTAGGTACTTTCTCCTTGTCAAAATACATTTTGTTAAACTTCTTGAGTAGGTCACCAATGAGCCA

[0293] AGGCTTACCAGCTATTAACTGTTTGCCTTGTAGCTCAAACACTTCATGTATTACTGAGCCAAGCGCTGTGTACTTATTACC

[0294] TTTTCGAATACCTAGAATGTAGGTTCTAAAGTAAGTGTGAGGGCAGCCCTCGTAAGAAGAAAGCTGCGAGTAACTAATG

[0295] TACTTCTTTGGGTACAACTCGATTAAATTCATCAATTCAACTCCTCGATTCTGTTAGCCATAGCTTCTCTCCCTGTTTTTC

[0296] TCCAGAGACTTTGACCACTTTGTTTTTGACAATTAAGTGCTCATACTTTTTAAACTGGTTTGCAAAGATTGTAACCTCAC

[0297] GTAGTCCCTGTAGAGTATCAATTCCTACAAAAGCCATTCGTCTGCCAGAGCGGTCATTAATTGTCTTAATAGATGTCACAC

[0298] AGCCACCAATAAGAGTTTTCTTGTAGCCCTCTGCAAAGTCCTGCCAGTTTTTGAAGTGGTATTTCTCAAGAGGGTGAGC

[0299] TGTAATGTACACGCCAAGTAAAGTCTTTTCATACTCAGCCATTATGTTCTCGTTCCACTCCTCAGCTGCTACCTCTGCAAT

[0300] TTTCTTTTTCGTATCACCTTTGAGCTCGTAATAACTTAACTAGTACTTCCAGGTCTTGTTAGGTCTGGCTCTATCTCGTCAAAC

[0301] GCTCCAGCAAGTATAAGCGGTCTCATAGGTCGAGCATTCACGACACGCTTGTTAATACGTTCCATGAAGTTCTCAAGAG

[0302] AAGTAAACCTGCCAGCTTCACGAGCTTCCAGATCCTTCAGAATACCTTTACAGCTCCGTCTCCTACTCCTTTAATTACTGAGAGAGGA

[0303] AATACAATCTTAGTGCTTGTAGCCATGAAAGTAAGACCAGACTTTATTAATATCTGGAGCTTGGAAGTCAAAGCCTTTCCG

[0304] TCTCACGTCTTGGAATAACTTGAGCTATTTGTCTAGGTCACTCATTTCGTACTCATAAGCGCTGCTTGGAAGTGAGTAGG

[0305] GAAGTGGTGCTCTAACCAAGCTGTCATGTAGCTTATCATTGAGTACGCTACTCCGTGGGACTTGTTGAAACCGTAGCCCA

[0306] TGTACTGTACAATTCGACTCCAGAGTTCATCCATGCAATTTTTAAAATCTTCACGTTGTTCCTCAGTTCCATATGAAGCAA

[0307] AACCATAATGTACTACTGAGTCGTAAATAAACTCATCTCTCATAGCTTCAAGCTCCTCAGCGCTTGCCTTACGTAGCTTAT

[0308] CGCCTCGTCCATAATCCCAACCACGAATGTGTGTACAATCTCCATAACGTGCTCTTGGAAGGTAATGACTCCAAAGGTC

[0309] TCACCTGTAATGTTACGCTCATCTGGGTGACTGTAGTGCTCTGCTTCCTCACCGTTCTTAATACGCAGCCAAGTA

[0310] TCTCCACTCGCTAGAGCTGGTGGTCTGTACAGTGCGTTTATAGCTACAAGGTGCTCGAATCGAGTCGGCTTAACAGATTT

[0311] GCAAAGCTGTTGCATACCTTGGGAGTTAAACTGGAATACGTTGGCTGTCTCGCCTCTAGCAATCTTAGCGTATACTTCTG

[0312] GGTCATTAGCCATTTTAGCTATATGCAAGATGTCAGGACGCTCACCAGTCTCCTTATAGATTGACTCCAAAGCTGTACCTA

[0313] TTACTCTCAGAGTCTTCAAACCAAGAAAGTCAAACTTAATGCCACCATACTGCTCAATCTCGTCTTTGTTCCATTGAGTT

[0314] ACAATGTTTCTCTGGTGCTCCATTTGGTCTTCTAAGAATGTTGGGAAGTGGTCTGTAATTGGTGTTGGACTTATGAGTAC

[0315] GCTCCTGCATGTTTTCCAAACTTCTCAAGCACTCCCTCGAATAGCTCAGCCATCTGAGCAATATCCTTTTCATATACCCA

[0316] GTCCTGTATATGTTTGTTCTCTTTAGCTTTCTTGCTGTTTCGCTTCATTTTCTTTGAGCTAGTTCTCAATTTGCGTAACTCTG

[0317] GGACTTGTGCGTATGCTTCCTCAAGAGTCAGTCCGTCTGGTACTAGATTGGTAATCCGTTGAGATTCACCAAAAGGAATG

[0318] TCATAAATAGCGAGTCATTCTTGTAAGCTGTTTTAGCTGTCATGTACACTGTGTTTACTATCTGTGCTACCTTGTCACGA

[0319] CCATACTTTTCACGTAGATACTCAATGAGTTCGTACCTTCTTACGTCCTCAATATCTGTGTCTATATCTGGGAATTTCAAGC

[0320] GAGTAATGTCTAGGAAGCGCTCAAAGTACAACCCATGTGGTATAGGTCAACCTCAGTTATATCTAACAAGTAAGCGACT

[0321] AGTGAGCCACCTGCTGAGCCCCTTGCTGGCGCTGTTATAATGTGGTTAGCTCTACAAAACTTGAGCGCATCAGATACAAT

[0322] CATAAAGTAATCTTGGAAGCCTTTCTCACGGATAACTCCCAGCTCAAAGTTAATGCGGTCCACATACGTCTTAGTTACTG

[0323] GGTGTACTTTTCTGAGTAGTCCTTCCTTGACAGCTGCACCAATCTCCTTGTCTATTGTAGTTCCTTCTGCTACTGGAAACC

[0324] GAGGGAGCAAACCTTCAACCTTCTCAAGTTTAAAGTCTACTTTGTCAGCTATCTCAAGAGTAGCGTTAAGAGCTTCAAT

[0325] CACTGGAGGCTTTTTCTAGTCCGTTCTTTATGAAGATACTAGCCATTTCACGAGCATCTTTCATGTAGTAGTACTCCTCACA

[0326] AAGATACTCTGGAGTATCTTCTGCAAGCATTTTCTTTCCACGACCTAAACAGAGCATACCTTGATGAGCAATTTTGTCCTC

[0327] TGCATTTACATAGTGAACGTCACCGCTCGCTACAAGTTTGACTCCAAGCTCTTTACCTATTTTGACTAGCTCCTTGTTGAC

[0328] GATGTACTGCTCTGCTTGTGGAGTAGGTTGTATTTCCAAGTAGAAGTCCTCTCCAAAGATGTCAACAAAACGCTTTGTAA

[0329] GCTTTCTAGCTTCCTTGAGTTCGCCTTTCAGTATCGCTTGAGGTATCATTCCAGCAAGACAGCTGCTCGTTGCTATCAGTC

[0330] CCTCGTGGTGCTTCTCAAGTAGTTCCCAGTCAGCTCTTGGTCTGTTGTGAAAACCAGTACGGTAAGCCTCTGAGCTGAG

[0331] TTTCATGAGGTTGCGCCAGCCAGTCTGATTCTTTACAATATATAACTGGTGGTAGTTCTCTGTAGTTTTACCATGACCGCC

[0332] TCTGACTTCTGTGAGATAGCCTTCCATACCTGCTAGTGGTTTAATGCCTTGTTTGTCGCACTCAGCAAACAATGAAGGAA

[0333] TCATATGGAGTGCTCCGTGGTCAGTCAGAGCAACAGCTTTTTGACCAAGCTCTTTAGCTCTTGCAACCAGCTTATCTACT

[0334] CTTGAGTGTGCATCTTTGAAGCTGCCACAACTATGAGTATGAATTTCAATGTAACCAACTGTTTGCTTTTTCTCTGCTACT

[0335] TGTAAACTATCAGTCATTTATATTCTCCTTTCATACCTTGGTGTTTCCTTGGTAATACCTATTCTATGCTATATCTTTCCAGAA

[0336] TGCAACAACTTTCTACAAAATATTTTAAAAAGAATAAAAAAGAGAAGGCTGTTAAGCCCTCCCCAGTTTCAACTCAA

[0337] ATGGATTCCCCAGAGGACTGTACTCTGGTTTCATTAACTTCTTAGCTCCAGCCATTCCGTGGTGCTTGACAAGCAAACTA

[0338] AACTGTTTAGTAAAGCGAGGTAGTCCATAGTAGTTCATATAGCGTCTTATAGTTCGTGCTCGCATTCAAACAACTCAGCC

[0339] AGCTCAACAGTGCTCATACCTTCTGTGTAGTAAAGACTTGCAAGCACAAAAGGGTTTTTGTAGCTCCATTCTGTGCGT

[0340] GCTTCCATCCAACTTCAACTAGTCGCTTTCCTAAAAGTGGTTTGCCATATCGCTCATATTACTTACCTCCAAGCTTGAAG

[0341] TTACCAATTCCATTGTCGTCAGACACTTTGTTGTAATCTCTCACATGTCCCTCAAAGAAATCAGCTTTGACCTCACTAGGA

[0342] TTCTCAAAGTATTTAATCCACGGTAAAGTATTATGAGCATCGTAGCGCTTCTCAAATCCCATCTGCATAAGTCGTTTGTTA

[0343] GCTGTAAACTCAACGTACTCATTCAAGTCTACTGAGTCGATGCTGTCAAACTTGTCACCAATAATGTAGTCGCCCCACTT

[0344] TTGTTCCAAGCGCACAGCCTCATCCATCGTAGTACCAACCCACTCACGAAACTCTGGAGGTGTTCAGCTCTGGGTTTTCTC

[0345] GTAGTGTTTCGATAAGCACATTGCCATAGAATCTAACGTGCAAGTTCTCGTCTCGGTTAATATAGTTTATCATTTCATTAGT

[0346] ACGACTCATTTTTCCTTGACGAGCAAGGTCATAAAAGAAAGCAAATGAAGCATAGAAGTTGATTCCTTCTTAAGATAATAT

[0347] CTGCCACGAGGGAGCGAATAAATGTCTTAGGAGAAGGGTTATCTATAAAGTCTTGGTATAGATTGAATATAAATTCATTAC

[0348] GCTCAAGAATAACCTCGTTAGTACGCCAGAAGTTTGCCACCTCATCCTGTACTTGTTGTGGTACTAATGAGCTCAACACA

[0349] TAGCTGTAACTGTGGTTGTGGTTTGCCTCCTGCTGAGCAATGTTAATAGCAATAGCTTGTAGTGAGCTGTCAGTAATATAC

[0350] TTTGTTGTTTCGTCCACGTAAGTTGTTTGAATACTGTCTAGTCCGTACAGTAAGCCAAGAATTAACTTGAAGCTTTCCTG

[0351] CTCATCTGCTGACAATGTGAGCCAGCTGGTTTGGTCGTCCCTCATAGGAATCTGAAACGGTGTCCAGAAGTTGCTGAGA

[0352] ATCTGTTTGTACAGAGGAAACGCCCACTTAAAGCGAACGTCGTCCCAGTTTAATACTCCAGAGTACTCACCGTTAATAAT

[0353] TCCAGTCGAGCGGTTTGGTGCTACCTTGTTGAATAAGCTTCTTTCGTTTAACATTTAAATTCCACCTTTTCAGTTTTATTAT

[0354] TGGCAAGCAACACAGTCGTCCTCAATCTCTTGAGGGTTCTTAGTTCTTACATAGTAAGTGGCTTTCAGTCCACCCTCCCA

[0355] AGCTTCCATATCACGCTCATACATTTCTTGAATCTGTACGCCTTTCTTAAAGTACATGTTGAAGCTAATCCCTTGGTCAAC

[0356] GTGTCGCTGAGCTGCGATAGTACTCTTTACGCTCCAAGATTGGTCCATATTGAAAGCACCTTTCGCATAGTAAGGGAATG

[0357] TAGCTGGCGATAAGTCTGGAGCTACAACAGTAAACTTGAAACCTTTTCGCTCCTCGCTGTACTCTAAGTTGAAAATAGG

[0358] GTCACGACTCGCTGTTGTACCTGCAACCACTGACGTTGAAGTGTTAGGAGCTACAGCCATAATGTTGCCATTACGGACT

[0359] CCGTACTCGTATACATCAGCTTTTAGATAATCCCAGTCAGTATCCAACCACTCTGTATGCGTATAGCCTCGTCGTTCAAAG

[0360] TACTCACCAGTCTCCCAGTCAGAGCCTTTTGAATAGCTCGTAAGCTCCACGCTCTTTAGCTAATTCCATTGAGGACTTAAT

[0361] AGCGTTGAAGTTAATCTCTCATACAGTTTGTCGTTAAACTCGATAGCTTTTTCTGACTCCCAAGGTATTCCCTCTAATGC

[0362] CATCATGTGGTGAAGTCCGTAAGTTCCAAGACCAGTAGCTCTGTATCTGTCGTTGGTATCAACTGCTTGAGCTACGTCTA

[0363] TGCGTGACTTATTGAGGTCGATAATATTGTCAAGCATTCTCATAAAACACTTTAATCGACTGAGCTAGATTCTTTCTGTTTTC

[0364] TCCAACAAACTTACTAAGATTCAAGCTTGCTAGATTACATACTACAAAGTCACCCATTTCAAATGTTAGCTGCTTCTGTCT

[0365] CTTACCGTTATCCTTGACAATAATCTCCTCTAGGAATACAGTCGGGCTCATGTTCTGGAAAATCTCTGTGCATAAATTCGA

[0366] GCAGTAAATCATACCAGCGTGCTTGTTAGGGTTCTTACGGTTAGCCTCGTCTCTGTAGAACATGTAAGGCTCGCCACCTT

[0367] CAAGACGACTCTTGAAAATCTGAATGTAAAGGTCGATAGCTTTTACAGTTTTACGAGGTAGTAGTGGGTTAGCTACACA

[0368] AGCTGCATAGCGTTCACTGAAAGTACCGCCTTCCTTACGACCTTTCTCGTCGTAACAATCTTGTAGTTTGAATCCCATGT

[0369] GCTGCTCTACCTCATAAGGACAGAATAAGTGCCACTCACCGTCTTCTCTAAGAGTCTTCATGAATAAGTCAGGAATACAT

[0370] ACTCCAAGGTGAATCTCATGAGCACGTTCTTTCTCGTCTCCGTTGTTCAATCGTAAGTCAAGAAACTTCTCAATATCTTT

[0371] GTGGAAGATGTCAAGATAAACTGCTACTGCACCTTGTCGTTTTCCTAACTGGTCCACTGAAACAGCTGTATTGTTGAGTT

[0372] GACGAATCCAAGGCACAACTCCACCGCTTGCTCCTTTGTATCCACGAATAGGTGCGCCAAGAGCTCGAATGTTTCCAGC

[0373] ATATACTCCCAGTCCACCGCCAGCTTTACTAAGTCGAGCAACATCAACGTTATTGAAGTAGATACCCTCAAGTGAGTCGT

[0374] CCAGTGTATCAATAAAGCAACTCGATAGCTGACCAAACGCTTTAGCTGCATTCCCCATAGTTGGAGTAGCCATAGTACCA

[0375] TTGAGCTTGCTAAGCTCCTCGTATCCCTCGATAATTCGAGCTAGACGGTGTTTTGTTTCGTCTTGGTGTATAAACATACTC

[0376] ATAGCCATCCAACGCTCTTGAGGGAGCTCCCAGACACCTTTCTCATTGTCTTGAGCACAATAGCGGTCTGCCAACATAGT

[0377] AAGACCTGCATAGGTAAACAATTTGTCACGCTCTGGTACGATAGCCTTTTGTAGTTTTTCCATTTCTTTGAAGCTAAATTT

[0378] ACTCAGCATTTCTGGAGCATAAACTCCTTTGTCAACTAACTCAAGAATCAATCTTTCTAGTGAGCGATGAAAATTATATAT

[0379] AGTTCCTCTGTTTTTGCGAGCTGCACTGTATAGTTGTTGTAAACGTAAGTTACGAGCTACAGCGTCCCACTGTTGAGACG

[0380] TAGTACTCACTTGCTCTAGAGCCAGCTGAATAGCTTTTTCTCTTATCGCATCAGTTGTAATGGAACGCTTACGAGTTACGA

[0381] AACGTTTCACTCTGTCCAGATATTTAGCTCCTTGCTCAGCTGTCAGTCCATCAACAACTAGTGCATCCATAATTGACTTAA

[0382] CCAGTTTACTCTCGTTGTATTCTTCCTCAATCGTTCCTCGTAAGATAATAACCATTACTTACCAACCTCCGTAAATTTAACT

[0383] TTTTGAATGTCACACTTAGGTAATACCAATGAGCCAACTTTTACAAAGTCATACTGAGCGCTCTCAAGTCTTAGAGCTAC

[0384] AACTCCTGCCTCGTGGTCACTGACCTCCATTTTAACTGGTGCGTCACAATCTCTCATATGCACATATACAAGTTTCATTCT

[0385] TCTTTCTCCCTTCCAAACAAGTCATACCATTCCCAGATATTCAAGCGATTGTCTGCTACTTCTTGGAGTTGTTCTTTCGTG

[0386] CAATCGTCAGCGTCGAGCCCTTCTGGGTAGACAAGACAATATATCTCAAACTTTCCTTTCGCTTTTTCAATTGCTTTCCTA

[0387] GTGCCATTTCTACCAGCTTTGTCATTATCGAATGCAAACTCAAGCGTCGTCACTCCTGCCTTGTAGAGCTGCCTTATGTGT

[0388] TCATCTGTAAAGAATATTCCAATAGGGCAAGCTACATTTTTATGACCACAATCCCACCATTTAGCTGAGTCAAATAGTCCC

[0389] TCTACAATCTTGACTGTGTTGTGTGCTTCTCGAATGTATTCTAAGTTTCGTCCAAGTCCTGTGAGAAAGTAACCAGTGTG

[0390] TAAATTCCTTGGTCGATGCATGAACTTTTCCACTTGCTTCTGCTTTGGTTGCTCTGCCAGTTATACCTACGAGTCTCTTGTC

[0391] TACGTCCTCAAGAGCCATAACTATACGGTCTTTGAGTTCTCCCTCAGTGCAATAGCGTATTTTCCAAGTCCAGTGTCT

[0392] CTGGTGAGTAGCCTCTGTAGTCTTTCACTTTAGCAAACTTCATGTTCTTAATCCTAAAAAGCTGGGAGGTCATGCTGCTGG

[0393] TGCCTCTTTCTCATTTGTTCTATGAACTTACGAGCTTCTTTTCTGAAATGACCTTCCTCAACGTGTTCAACTGTCCAGTCA

[0394] ACATCTATTCCAAGCAAGTCAGCAAGCCAAGCCAGAGACTTCTTGAAAGTAATCTCCTCGACTTGCATTATCAAATCATA

[0395] AATGTCTCCACCATCTAGACAACCTGTGTGACAAAACCAGTTGCCAGTTGTCTCATTGATTACAAAAGCTGTGGGGTTG

[0396] TCGCCACCGTGTACAGGACAACAACAACGCAACTCCCCAGAGGCCTTTAATGTGTTGAGCTCCCAAGGCATGTAGCACCT

[0397] TGAGAACGTCAAGTTCTTCTCTTAGTTTGTCAGCAACACTCATAGTAAACTTTTCACCACACAAGCCAATAAGATAGCA

[0398] ACAAGTACCCAGCGCATCTTTTCAGCAAATACTTTGTCGTTATTACAACGATGTCCAAAACGCAATAGCCCTCTCCAGAT

[0399] AGCAAACGCTGTAGCTCCAATCAACGCAATTACGATAGTGTTAATCATAAAGTAAGTCACTACGCAACATCCTTTCTTGG

[0400] TGGTTTCTCCATTGACTGTCTCCAGTTACCTCTCTCGGTATACACAGGTGTCCAGTCCATATCTTCACCCATTCGCTGAGC

[0401] TATAATTTTAAACTGGTGAGTTGCACCTGTCATAGCTTCCTCGTCCTCGCTTGACTTTCTCAAGTAACAAACACGGTTTAC

[0402] AAGCTGTAAGATTCTATCTGAGCCAGCAATGTCTCCCTCGTCCATTTCGTCACCCTTACCGATAGCTGCACGACCTAACT

[0403] GTACCGCTGTAGCAATTGGAATCTGTAGCAACCCTGCAAGGTCTTTTAGTCCACTTGTCAGGTAGCCAAGAGCTTGATAC

[0404] TCCTTTTCACCCAAGTTGCTGTTTGAGCTTGGTAGCTTAATGTAATCAAAGACTACAAGTTGTATACCGTGTTCCATCTGG

[0405] TGCTTTCTAATTACGCTGGCTATCTCACCGAGCGTAAAGTGTGGCATGTATATGTGGTAGAAAGGAGCTTTCCCTATTGCT

[0406] TGACTGGCTTTCTGTACAGCTGCTACTTTGTCTTTTCCGTTACCGTGGTGAGTGTTCTTTCCAAAGTAACCATTACGTATC

[0407] TCTTTGTGCTGTACTCCACTAATCATACTGAGGAGTTTGTCTTCTTGCTCCTCTTTGTACATTTCTGTGTCGATATACAGTG

[0408] TTGGTATGCCATCTTCAACGCAAATCTTTTTACACCAATTTAGTAACGTTGTAGACTTTCCGACCTTTGAGCGAGCACCTA

[0409] CTACAGTCAACTCACCGACAACCAAACCCATTGAAGCTAAGTCGAAAGCGTGCCAGCCAGTTTTTAATCCAATGACCTC

[0410] AGTAGGGTTGAGTAGTCGCTGCTTTAACCTGTCAGCCATACCCTCACCGAGTTTCTTGACACCTTGAGCTACTTGGTACT

[0411] CCAAAGCTATTTCTCTGTAGTCTCCCTCAACAGTCTGTAAGAACTCATTGAGAGGAGTCTCTGCTGCTTTGACTGCTTGC

[0412] TGCATTGTCTTCTGAGCTTGGTCGTATACTGCTCTACGAGCTGCTGCTTGCTTTATCTGGTCAATGAATAACTTGGCGTTT

[0413] CCCATTTCTGGTTGCAACTTAATGCCTTCTAGGTAGTCAATTCCACCCAGCTCCTCAATTGACTTCTTGGCTTTCTCATCT

[0414] TGAAAGACTGACAATATTGAGATAGGGTCAAGCGTGTTACCACTCTCAGTCAAGTGAAGCATACTCATATAAATGTAGCG

[0415] ATTACCGTCTACTGCAAACATATCAGGAAGCAAGCCACTAGAGCTGGCTGAGATTATCTCAGTAGGGTCATTTAAACAAA

[0416] TCCCTAGCAATGCCCTTTCCGAACCAGCTCGGTTTATTAAATCTTTCGATGGAATTTTTTGAGTCATACTTTTCTGGAAGC

[0417] TCCTTTCTTTGAGATTCTATTCTTGCTGCTACTCTCATAGCTTTAACTATACGCTCTTGTTGACCTTCAAGCCCTTCCAGTA

[0418] AGTTCTCGCAATTGAGCTTTAACTCTTTAGCGTGAGTTATTGCGAAAAAAATTTGCTCAACTGACCAGCCCTCACTAGTC

[0419] AGCCAAGCAACTTGAAAGTAAACATTTGTCTCGCTATTTGGGTAAGCTTGCGAGTGAGCTTCAAAGATATTACTCAAAA

[0420] GAAATATCGTTCAAGTCGTCGTCTGAAAGCTCCTCAGACTTCTCAATATTCACTGGCTCATCTTTACGAGCTGGCTTTTCT

[0421] TCTTTAGCTGGTGTGTCCTCAGTATGACCAGCTGGCGTGTCTGACTCACTAGATTCACTTGAAGTGCGTTCCAGTACATC

[0422] GTCCCACTTAGCTCCGTCAGCTACACGCTTAGCTTGTTCTGGAGTACAGAAATCTGCACCTTTTAGTTTTACTAAGTCAA

[0423] CTTTGACTTTGTTGTACTTCTCAAGCACATCAGCTGCAATCTCCTTGTCTCCACCCTCGTACATTACTGTATACTCAGTATC

[0424] TTGGAAGCCAGTACCCTCACGGTTAATTACTAAGTCGTACTCTCGTGGGTCGCCTTTACGAGCTGCAATCTTTTTAATATT

[0425] TTCAAAGATACCGCCAGTACCGTCTAGTAATTTAATCTCACCAGCTGCAACGTCCAGTACATGAATTACATACTTAGTTTT

[0426] TAACTTGCCATATGGAGTCTTGCTATATCCGTGGTCACGTAAGAAATCCTTACGAGCTTTGTCTTTCAAGCCTTTTGCGTC

[0427] AGCTTCTTTAAACTTTTTGTTCATAAACTCTTGGTTTGCTTTCTCAAGTAAGTCTTCTTTACCGTGGTAAGGAATACGACA

[0428] ACCTTTACCGTTTTTACCACCACCGTCACCTTTCACAGCCCACCACTCATTGTAGCGGTGCGGTACAGCGTCCAGAATAC

[0429] GAATCAGAGTTGTCACCCTCAGCTAAAGTTAAGAAGTCAATCTTATTTTCTGGGTTGAAACCTCCACCGTTGTTTTCCTCT

[0430] TCACGTACTACTGAATCCCATCCGTTTGCATTAGTCATTTTTACATTGCCCCTTTTCATTTTATTTGAGCTAACTAGCTCGC

[0431] TTGGTTAACTTGGCACTGACTTGAGTGGCTGTGTTTGCCTCTCTGTCTATTATATTACTATGGTCATACTTTGGTGTCAACA

[0432] ACTTTCTACAAATTTTTTTAAAATTAATGTTGAGCGTTTGCTGGAGGTGGTGACTCCCCAGCTGGCGGCTCTGGTGGTAAT

[0433] TGACTTTCTAGTAACTTAATAGTAAATTCTAAGTTGTCAATTTCACGTTGCTTGTAAGCAATATCTTGGTTTTCTCTGTCAA

[0434] TCTTTGCGTCGCACTCAGCTTTTACTTGAGCCAGTATTTCCTCCTGCTCAGTAATACGGTGACGACACATTGCTTGAGCG

[0435] TCTTTCAAGTTGCGTACGTTCAGTTTCAATTGTTCAATAAATTCAGTCGTTACTCCCATTAGCTTCTCCTCCCAAGTCAAA

[0436] TGTAATAGTGCGAGCTTCTTGAACCACCAACAACTTGACTTGTATCTTGTTGTCAATGACCTCAACGTCGCTCACGTAAA

[0437] CCTCCACCCCATCAGCTTTCATTTGCTCAGTGACCTCGACAATTTGCTTTGTGAGTTGCATCCTCTGGTAATGAATAACCG

[0438] AGCTGTCTAATCAATTTGTCTTGCTCAGCTGAAATTAAGCGTCAACCTCCTGCATAGCAATGTCGAAAGCTGTAGGCTT

[0439] CAAAGCTGCTTCCTTCTTAGCTCGTAACTCATCAATATTGAAGCGCTTAGCAATTTCTTTCCCCTTCATGTTGTTGCCTCC

[0440] CAGTATTAAAATTGTTGTACTATTTAGTATACTAACTGGTAATACTACACAAGTCAACATTTATTTACAAGAATTTTCAATAC

[0441] CTTGGTAACTCTCAGGTCTTACTACAGTATATTGTCGAGTTACTGAATGGTTACAACTATTTATGAAAAGGGTGTTTATAC

[0442] AAAAAGAAAGACTGACTCCATGAGCCAGCCTCCCAGTACTATTGAGCGAACAGTGTAATTATATCTGTTAGTATCTGCAT

[0443] TACTTTTTCTGCTTGTTCTGGCGCTACAGCACCGCCAAGAAGTACGCCAGCAATAGGGACTAACAGGTAAGCAATTTTTA

[0444] AAGTTTTCTTCTTGTCGAGTTTCATATTATCACCTTTTTATCCTTTCATTTTTATTTGGACCTGCCCTCAGTGGGACTTACG

[0445] GAAAGTGTTACTGGAGTATGACTATTAGATATACATGAACCACCAACCACGAGCTGCAAAGTAGTTGTCTAGCTTCCAGA

[0446] TTTTCTTTTCGTCAATGCGGTCAGTTACTACATAGCAAAGACCGTCAAGTTGAACTACTAGTTTACCTGTCACGCCAAAC

[0447] TCAACCATTTTACCCATAACCGTTGGAGCTTCTGAGAATCCGAAACCACCAGATTGAATCGCTGCTGTATTCACAGTTCC

[0448] ACCTCCTGTGCTACCTCCACCGCTAACAGTTTGACCTGTCAAAGCATAAACGATTGAGTTAGCTATTTTGTCAACGTTCC

[0449] ACTTAGCCATATCTCCCTCATTGTCAATGAAGCCTAGCTCAAGCAAGATAGCTGGTGCTGTTGTGCGAGCAAGTACAGCT

[0450] AGTTCTTTTCTCTCTTTAGCGCCACGGTTAAACCAACCAATATCTTTAGCCAGTTGAGCTGACACCTTTGCAGCTAGAGC

[0451] TTGTTGGTCGTAGTAACAAACCTCAACCCCATTAGCTGAACCGTTGTAAGCGTTCAAGTGGAAAGATACTACTAGGTCA

[0452] ACTTTGTGGCTATTGCACTTAGCAACGATGTTATTTAAGTTAGCATTCTGAGTTGAACCTACCTCGTCTGTATCGTCATATA

[0453] CTGTGTGTCCAAGAGCTCGTAGTTTCCCTGCTACTGCGTCTTTAACTTGTCGGTCCATAATGTGCTCCTTTCTGTTTCCAG

[0454] AGTTAGCACCTTGAACTATTGAGTTATGACCACCATGTAATGAGAAAATCATACTATCGCCTCCTATTTATTGTTTTGGCA

[0455] AGCTATCAGTGTTGCCCTCTCCAACCTGCCAGCCTTCTATTTGTTCAACTCGATTTTCGAGCTTTTTTTAAGCTATCCTGTAT

[0456] TTGTACTTGTGTCTGTTATGCGCTCCAATTGGTTTGTGTTCTTATCAAGATGCTTCATGAGTTTATCTTCTCGCTTAGTT

[0457] GCGTCTTGCAAGCTTTTCTCATAGATGCTATGAATCTTTTCTTCTCGGTCATTGTTGTCAGCTAGTAGTTGTTGCTCCCTCG

[0458] CTTTACTGTCTTCTTTTTGTTCCTTAATAAGCGGACTACCCACTTGGCTACTCCTAGCAAACAAGTAATAAACAATACTG

[0459] CAAAGATGTACTCACCTTTTACAGCTGCAATAATTTCTGGGATTCCCACCAGTCTCCACCTCCCCTCATCAGACTATCTTTC

[0460] CAATTATCAATATAGTACTCTGAATCTGCACGCAAAACACTCTGTCGTTTGCTGCTGGTGTGTAGCTAGAAAGATAAGGG

[0461] TAAGTTACATTCGAGTCAGAAGACGCTCCAGCAAACTTCAAGCGAGGTCTGCCAGAGGAATGAGTGGAGCTGACTGTT

[0462] GCAATTACGAAAGGGTTTTTATTCTTGTCTTGAGTCATACCAATTATCAAGTCAACAAGCTCGTTAGAGTCCATCATCTTA

[0463] AATACACCACCCTTCTAGCCTCGTGCTCCATTTCTCCACCTGCTTTGAGTTGCATCTTCCAAGCTGTCTCACCAAACGTG

[0464] TCGTCAATGTCTAACGAGCTGTAACGAATGTGTAAAATGTCTTGGTACTCATGCATTGGCATTAGAGCTGTCTTGAAGCG

[0465] TATCTTACCATATGAGGCTGTACTTTCTTGTATTAAGCGGTCAACGTAAGCGTCAAGAGCTGCTTGAGTCTTGATGTTCTC

[0466] AGCCTCTCTGTAGTCCACTATAATACGTCCCACTGACTTTATAGATGTAATGCCTTGGTCGTTATTCTGTCGGATTCTCGTA

[0467] AAAGTCTGGTTACCGTCAAGGTCATTCGCAACAGCAACCCAAGCGTTAGCTGTCTCAAATAAATCTAGTTCGCTTTCCAT

[0468] ACCGTTGTACGTTACAGACAACTCATCATCAAAATACTTGTAGTCAGCTGGTCGGCTTGCTGGTGAGATATAAGGCGAGC

[0469] TTCTAAAGATTCCGTTAGCATCAACCCAAAGAGGATAGTAGCCAATAGCTTCAAGCATTGTGTTAATCATTTCCAGATAA

[0470] GGGTGTTCCAATTGGAAACGTAAAGTTTTGACTTTGTTGAGTTGGTGGTATACTAATGAGGTTGGGGTCAATGAGTACGTC

[0471] ATAGTTTGTCATTATAAGACGGATATTCTGTACAACGTTTCCTGTATAGGTATGATTCCAGTAAGCCTTAATGTCCCTCAAA

[0472] ATAATGAGCTGGTCGTAAGCATCAACCTCTCGGTGTACCTTGTTAGTATCATCTATTCGAGTTGGACTGTTGAGCAAGAA

[0473] GACTCCAAGAGGAAAGTCAATCCAGTTTCCATCTTGCATTTCAATCTCCATGTATGGTTTCAAGCGGTCACTCGCATAGT

[0474] TAATGATGTCCGTTTTAGGTATCAATTGGCCTTCCTCATAAATCATTGACACTTGAACATTTCCAAGCCTTACGTCGTCTTT

[0475] ACTGTTCCAACGAGACATACTGTATTGTAACTGAACTCTCAAGTTAGTTGCATCAGTTCCAGCTGGCACGAGTTTTCCAT

[0476] AATAACTTACATCTGTCCAAGCGCTCCAAGTAGTACCTCCATTGGTTGAATAACGAACTTTCATACCCTTTGGTGGCATCC

[0477] AAGAGCCTTCACGAGTTGGCGTTCCAGATTTGTCTCCAAAGACTAGGACACTATCTTTTACTCGTGCTACGTCGCTACCT

[0478] AGAGTAGTAGTGTTGCTAACATCAATAACTGGACTAGTAGCCATACCGCTATAGATTGGCTTGTTAGTTCCAAAATAGATA

[0479] TTGTCATAGTTACAAATTCCATTATTGACTGCTGAGTCACAAGCGATAAGTAAACCATAAGTACCAGTAGCTCTACCAGA

[0480] CGGACACGTACCAACAGCGTCACAGCGAACCCAGTTGTTATAACCTGTAGGTGTATAAGTCACGCTCGTGTCAAGTCTA

[0481] AAGTTTCCAAGGTCACCCATTGCGTACAAGTAGTTAGGATTCACCTCACCGTTAAAAGTTCTAAAGTAAAATGAAGCAT

[0482] AAACTTGTTGTCCACTTGACAGTGAAACTAAATTGTTTGTTGCGTTGTAGCCGCCTCCAGAGTCAGTACCAGTTTGCATC

[0483] CCTACAGGATACTGAGTAGCACCCAAGCGAGTTACCTCTTGAGCCCAGCTTCCTTGCCAGCCACCACCTTGAGCTACTT

[0484] TGTACTTTGTCGGACTCCACCAAGGTATGAAACCGCCAGCCATCCAGCCATCTGCATAAATATTCACAGTACGCCAACCT

[0485] GTCTCAACGTCATTATTTGCGATAGTCCCATTGTAAGGCTGTAAAATTAAATCTCCACTAGCTGCATGTACTACAGCGTCA

[0486] GTCTTTACCCAGTTGTTGAAACTAAAAGCTGGGTCTTCTGCGATACTTCCAAGTTTAGTAATGACTGTTCGCTCAACTCT

[0487] ACCCACATGAGCTGGAATATATTCTTCCTCAATTTTAAACTTTCCAGTACGCTTGATAGTGGCAAAGGCAGCAAAAGCTA

[0488] CCTCGCCCTCAAGCACATTCTTTAGAGTTCTTTTATAATTGAGTTTGCGGTCCAGCAAGTCATATCTAAACCGCACATTTC

[0489] TTGGTTTATGCTTAGCATGAAGGACATCCATAACTTGTTGAGCTGAATATCCTCCACGAGCTAATGCTTGCATAGCCTCTC

[0490] CTCCTTAGTCCTCATTTTCCCAGTTAACATTTGGGTCATAGTAAGTTTGTGTAAACTCACAGCTTAACTCGAACCCAGCAA

[0491] TTGGTTTATCTTTAATGTCCACGCTCTCACACGTAGCATAAAGCTTACGACCTGCTCCGTCACGGTAAAGGTACGTTTTTC

[0492] TGGCTTGTAGTAACTGCTGAAACGCTTTAGCCTCATTCCAAGTATCTACCTCCCAGCTAACTTTTACCGTCGTGAGCTCG

[0493] TGCTCCCGTATTCTCTGACTGGGTTAGTTCTTCCAGCGAACCTTTGAAGCACGCTTTCTACCTCGCTATTGCTGTCTCGT

[0494] GCTGTGACCATTGGCAGAAAGTTGAAGTCGCTAATTTTCCGTACATCTTGCAGTATTGTCTCATTAAACATATGAGTGAA

[0495] CTGTTTACCTGTGTTGTCCGTTGTTCTGTTGTTGACCAAGTTCCAAGCTCTAAGCTTATACTCATAGTTTGTATGACTTGC

[0496] TGCGTGTAATCTAGGAAACTTCCTTGGATTCCGTCTGGCTTCATTGGCTCAAAACTAATAGTCTGGTCTGTAAAAGTTC

[0497] CCTCTTTCAGTGTGTGTTAGCAAAATAAGCCCTCGTGTTTGCTACTGACTGTCTAAAATAACAACGAATAGACGTATTG

[0498] TTACCACTATTGAAGTTTATCCATCTACCAGTTGTGTATGGAGCTATATTCACTGTGGCATCTGCATTGTAAATTCCAATCA

[0499] TTCCACTGTGGTGACTATAGAAAGTGTAATTTGTGTTAGGTTTAACATTTACTTGTATACCCCAAGCCATAGTAACTTCAG

[0500] TGTGTATAACACTAAAGTCATAATCACCATAGAAAGTTACACCAGTTGGGTTAGTCAAATTGCCTTGAGCTTTTGGTAATA

[0501] AGTTTGTAGTAACTGAGCTAGTAATACCATTAGAGCCTACTAGTGTCCACTCGTTGTTGCCAAGGTCACTGTACTCTCGA

[0502] CGATAGCATTCTATCAAGTTAGTAGCTACTGGAGACGTAATTGTTAAGCCAATACCAGAGAATATTGTTTTGTTTCCACCA

[0503] GTAATAGGTGAGTTTTGTAAGTTCTCAACTCCTAGTCTTATTTTTACTGTGTTAGCTGGACAGACACCACCAACAGTAATA

[0504] GCGATTCTGTAATCAGTTGTACCCCACGTTGGAGCTGAGTAACTTGAAATAATAGTGCCAGCGCTGTCTAGGTATTGCAC

[0505] GAAAGCTCTTACTCCTGTCTGGTTTGTGTACGCTGAGACATTGTAAGTAGCACCAACTACGATAGGAATTTGTGTTCCAT

[0506] CAAAGTCAGCCCTTACAAGTGTAGTAGCTGTTCCTACTGCTAACCCAGTAGTATCGAACTCGAAAGAATTTGCTGTCAGT

[0507] GGTCTTGTTCTTGCTACGTTAGTTACAGCACTCCAAGGAGCTGCCCAGTCGCTTGTAGTTGTGCCTCCCAGTGTAAATGG

[0508] TGGTAACTCAACTCCTAGCTCACCTTGGTTGTACACTATGTTTGTAACTCCTGCTCCTGCTTCCTCAATACGTTGAATGAC

[0509] TGGTACAGCTGGAGCTGCATAGTTGACCGTGAAAGGAGTTGTTACGAAACTTGACCAGACACCATTCGAATTAATTCTA

[0510] ACTTGCAACTGGTAAGACGTATTTGTTGCGAGCGTGTAATCAATTTGTACAGCTGTTTCGTTCAAGCCAGACTTACTCCA

[0511] CAGTGTCACTCCACTAGAGTTCTTTAGGAATAACTCATATTGAATCTGTCCTACAGTCGTCCACTCGGCTACCATTCTAGC

[0512] TGAGCTGTAAGTCATGTTGTAAGTTGGTACTACCATGTTTGGAGCTGCACTCGGATTAACTGCTACAATCATAGACTCTT

[0513] GAGACCAGTCGGAAGTCATCATAAACTCATCTTGAACCCTTACAGCCCACTCAATATTGCTTGCTGGTAAAGTACTAGCT

[0514] GCTAGGTTGTAATACTGGCTCGTTGAGTTTGCCCACGTTCCTCCTGTTGGCAACTGATTCCAGCTCCCTCGTGTACCATTA

[0515] GCGTCCACAGTTCTCCAACGGAATTGGTAACCGTACTGAGCACTTTTAGCGTCAGCCTTCCAAGTGAAACTTAAAATAC

[0516] TTGTTCGGTCAACTGTGCGTCCCATTTCTGGTTGCTGGTTTACTGGAGCTTTTGGTGGAACGTTATGGAAGATTGTGAAG

[0517] CGGTCCGAGTAGTTCCACTCTGAGTAATATTCTCCGTCATATGCTCGGATAGCTACAAGAGCTTTTGAAGTCTGAGCCAA

[0518] TTGGCTTGTGTCAAAAGTATACGCTGTAGCTCCTGCTGCTGGGACTTTAAAGTTTTTCCAAGTATAGCTATTGTCGTCTGA

[0519] GATAGCTACCTCATAAGTTATAATGTTTGGGTTTGGCTCGTTTGTTGGTGGTGTCCATTTTAGCTGAACTGAGCCATCAAC

[0520] CTTAACGCCAAACGTTGGGTAATCTATACGAGGTCTGTCTGGTGGAACGTTCCAGTCACCTTCAATAACCCAGTAAGCTC

[0521] TATAAGGGTTGTCAATAACACCCCAGCACTCACTTGCAGCTGTTCCTATGTTGTTCCATAACACAACCCCTTGGTACAAA

[0522] CCGCTTAGTAAATCTGCTTTAATAATATTCGTAAAATCAAAGTCTTGGTAGCCACCAGTAGTAGGTGTAAACTCTGTCATA

[0523] TCCTGTGAGTAGAATGGACGTAACGTCTGCACTGTACGAGTCGCTAGTTTGTGCTTACCTACTCGGAACTTACCTCCATA

[0524] ATAGAAGTAAGCTCTCAAGTAGATTTTTGCAGGTGTCTTAGAGGTAGCAAGTAGAGTTCGTACCTCGTCTGGTGTACCTA

[0525] AGAATGAAGTATAACCCTCTGGACCACCAGCACGAATAGGTGACCAACCACCATTCCACCAGTTATCACTCAAGCGCCA

[0526] TTGCTCCATCCAGTTAACAGGTATGACCTTCTTTTCAATAGCCATTTGTCTACCTCCTTATTTTCTTCTAAATGCTTGCTTT

[0527] AAATCTTTCCAGTACTCGTCTTGCTGCTCTTGCATTTTTTGAACTGTCAAGTCGAATGTTTCTTTGTTGCCTCCGTCGCCT

[0528] TCTACCTCAATCTTACCTGTCATGACATGCTCGTGAGTAACAGTCACTTGTTGTCTTCCTCCACCTGTGAAGGCTTCAAG

[0529] TCCAACAGTTCCGAACTCTGCTTGCATTTGGTCGTTCATTTGTCCCATAGCTCCTGCGATAGCTCTTGTCATAGGTCTCAC

[0530] TTGAGTTAGCGCTCCCTCGTACCAAGTTGGGAAGAATGACTCACCAGATTTGTCTAGGTCGCTCAGCGGTCCTTTCTTGG

[0531] CAGGTGAGAATGGTAAGTACTGTCTGACAGCATCCATACCGTCTTTAACAGCGCTTATAGCATTTCCTATGGTGTTCTTTA

[0532] TTCCGTCTACGAAAGCGTTAATAAGCCCCTTACCAGCGCTTAAAAAGCTACTTGCAAAGTTCTTAATAGAGTTGACCATG

[0533] TTCGTTACACCGTTACTTACAGCTGTCTTGGCGTTGTTCATCATATTGTTGAATGAGTTCACTAGGTCAGTAGCTAATCCA

[0534] CGAATGTAACCCATGACTTGGCTTGGTAGGTTGCTAAAGAAAGTTTTAATGTTATTGACTCCGTTGTTAACAGCGTTCTTT

[0535] GCGTTGTTCATCATGTCAGTAAACCATTGTTTCACAGAGGAAGCCATTGAGCTCACTGTAGAGCTAATGTTACTTTTCAT

[0536] GTCAGAGAACCATTGTTTAACTCCGTTGACCATATCGTTGACTTTCTGCTTAGCTTGTGTCACCATGTCGTTAAACCATTG

[0537] TTTAACTTGGCTACCCATCTGGCTTGCTGAGTTGCTTGCATCTGACTGCATATTCTTGAACCAGTTTTTAATTGAGTTAAC

[0538] CATGTCAGTAACCCACTGTTTAGCTCCTGTTACCATTTCGTTGAACTTGTTCACAACCCACTCTTTCATTTGACTAGCAAG

[0539] GTTACTCAAGTTAGTTGGCAGCTCACGGAAAAATGTAATAACTGCTTGCACGCCTTGTTGAGTCCAAGTCTTAACAGTTT

[0540] CCCACATGTTAGTAATCGTATTGACTACCCAGTCTTTCGCTTGAATAGCTAAGTTGTACAGGTTAGTTGGAAGCTCTTTAA

[0541] AGAACTGTATCACTATTTGTATACCATTCTGTACTGCGTTGGAAATTCCGAGGAATAAACCAATGAATACACCAATAATGA

[0542] AACCTACTCCGTACACTAAACCGTTAACAATATTCATAGGTAAGTCAGCAAAGAATTGCATTATAGCTCTTACTCCGTCGC

[0543] CTACAATCTGGCTCGTTGTTGTAAACAAGTTGTTCCACCATGTAGTGAACTGCTCCCATAAGCCCTGTAGAGTTGACAAG

[0544] ATTCTTTCTGGAAGCTGAGCGAACCAGTTACCTACCTCGTTAAACCATTCACCAATAGCTGTCAAGATATTAGTCAAGTT

[0545] TGTTGCCCAGTCAGTAAGCAATTGAGTTAAGAAATTAATAGTTGCAGTTTTAGCAGTTTCCCAACCATTTACAAACCAGT

[0546] TGCCAATATCTGTGAAGAATTGACCTACAGTTGTAACTATGTTGTTCCACCCATCAGAAATGCTAGTACCTGTTTCGTTGA

[0547] GCCAGTTCATTGTTGCTGTTTTTGCTGAGTTCCACATGTCAGTTAAGCTCTGAACTATGTCAATTCCGTAAGCTTTAAGCG

[0548] TGTTGTTAATACCGTCCCAAATTCCACCAAAGAACCCAACTAGTGCGTCCCAAATAGATTTAAAGAAGTCAACAGCAAA

[0549] CTGTGAAGCATTGTTCCAATGGTCCATAACACCTTGCCAGTCTCCACTTAGTAGAGCACCAACCATTTGAACCATTTCGC

[0550] TCACCCAACCAATGACATTCAAGAAAGCATCTAAAAGCGGTGCTATTGCGTTAACTACTCCGTTAAACACTGAGAGTGC

[0551] AATTACAAGGAATGCTCCTAGAATAGCTCCTACTGCTAAGATAATTGGCTCTAAAGATTTAAAGACTTGCTTTATTTTGTC

[0552] CCAGATTGGACCGACTGCATTAGCTAAATTGTCGAATGTGTTCTGGAAAGTCTGTTTGAATGCTTCAACTGCTGGGAGTA

[0553] TTGCTTGCCATACCTTGTCAAAGAAAGGCTTGACAGTTTCCCACTGAGAGATTACCAGAGCTGCTACTCCTGCGATAGCT

[0554] ACCCCTGCTGCAATAAATGGAGCTGCTGCTGCGAGCGCACTCAAGAAAGCTGGAGCTAATAGAGCGATAGCTGAAACG

[0555] ACTGCACCAATTACAATTACGATAGCACCAAACCCAAGAATTGTAGCAATTGTAGTTTGAGCTGCTGGAGAAAGGTCAT

[0556] TGAACCAGTTAGCCAAGTTCATTACACCTGTAGCTACTACGTTCAAAGCTGGTCCTAGTGCGTCTGTTAATGCACGAGCT

[0557] GTAACGTCGAGAGCTGATTCCATTTTGGTCATAGCTCCTGCCCAGCCATCAAGCATAGCGTCTGCTGCCTTCTTAGAACT

[0558] ACCTCCAGAGTTGACTAGCTCGTTTGTAAGCTTTTTCAACTTCTCTGGTCCAGCATCGACAAGGCTCATCATAGCTGATA

[0559] CTGCCTCAGTACCGAAAATAGTAGCTAAAGCTGCACCCTTCTGAGCTTGTGTCATTCCTTCCATTCCTTTGCCAAGCTCT

[0560] CCAGTAATCTGAGCTAGTGACTTCATTTTACCGTCAGAGTCAGTAGTTGAAACACCCAACTCTCTAAGCATATTAGCTGC

[0561] TGCCTTTGGAGGCTTAACTAGTCGTAACATTGCAGACCTTAGAGCTGTACCTGCTGTCTCACCTTTAATACCTGCGTTGG

[0562] ACATAATACCCACTGAGGCTGCAAGTTCTTCCATCGAGATACCAAGAGCGTTGGCTGGTGCTGCTGCATACTTGAAAGC

[0563] ATACTGCATATCACCTACTCCTGCTGCTGTAGAGTTTGCTGCCATTGCTAAAACGTCAGCTACGTGAGAAGACTGTGAAG

[0564] CTTCCATACCGAAAGCATTTAGCGCTGAGGTGATAGTGTCTGCTACCATTCCTAAATCCTCACCACTGGCTGCTGCTGCG

[0565] CTTAAAACTCCTGGCAAAGCTTTTGTAGCTTGAGCTGAGTCAAAACCTTTAGCTCCTAACTCTGCAAAAGCGTCAGCTA

[0566] CTTGAGTGGTACTGTATACCGAGCTCTTAGCCATATCTAAAATATCACGTTTTACTGTTCCGTAACTGCCACCAGTTAACA

[0567] CTGAGGCTCTACGAGTTGCTGCCTCAAAGTCTCTTGAACCTTGAATCATAGTAGCAAAGCCACCAATGACTGAGCCAGC

[0568] CAATTGACTCATAGCCATACCACCAACGCCTTGTAAAGCAGCCATACCAGCATTAGCTCGACCAGCTGTAGTTTGTAAGC

[0569] CATTAGCGCTTGACCTACTCGACCAATCTGAGCGCTTGCGTTGTCTCTGGCGTTAATTATATATGTCAACAATATTAGCCA

[0570] TCTACCTAGCACCTCCTCCGTACTTAGCTTTCAAGTCTCCTCTCTTAGCTTTACTCTCTTGCTTTTTAGCTTCATTTTCTTG

[0571] GAAGCGATTGACTGCGTTGTGTACAACCAGTAACTCCTCCATAATTCTATTGTCCTCATTCAGAATGACACTTGGAGGCT

[0572] GGTGGAACTTAGCGCATAACTCGTATAAAGTCAAACAAAGGGAGCGTGCTCAACAGTTTTACCTGCTAGGCTACGCTC

[0573] CACCGCTTCCTCAATTACTTTTTTTCTTGGTCAGTGACAGCATTGCCATCTAAGAACTTACCAACTTCTAGAATCAATTGA

[0574] GCTACGAACTCCTCATCAAGAATGTTGTCGAACGTGTTTAACTCAAGTGGTAAAACGTTGTCGTCCTCATCTGTAACGTC

[0575] CCAAGACTTAATAGCTTTAATCACTCGTAAGACTCCAACTTTTGATTGTCAGTAAATACTGCTTTACCTTTCTGAGTTGT

[0576] AGACTCTCCAACGATTGAACGAGTTTCACCGAAAGATAAGTTTCTCAGTTCGATAGTAGCTCCCATTACCTCAATAACGT

[0577] CACTTTTCTCTTTTCTTAACCATGCTTTTGCCATTTATAATCAATCCTTTCGAGTTTTAGTTTTTATTAAGCAAAGTACTTGT

[0578] CGATAGTGTCTTTCACTTCAACAGTAATACCTGTGTAGATAGCCTCAACAGACTGCTCTGTTAAGTCTTCTGCTCCAGCG

[0579] CCAATAGCGTTCGTGTCGTACTTACCACCGCTTAGAGTGAACTTCATAGCGTTCTTAGGGTTTGAAATATCTTGGAAGTTT

[0580] AGCTCAACAGTAATGTCTGCACCGTTCTTGAAGTTAGTCCATTGAGTTGTATCCATGAAGTCGAATGTAATACTAGATGTA

[0581] ATTGTCACAAGCTCCCTCTTGTACTCGTGCTGCTGTGTCAGTTCCGTTAATAGCAAACTTTCTTTCTAAGTTGTTAGCAATC

[0582] TCTAGCTCAAACTCTGAGATATTACCAACTGCTGTGTTATTAATCTTGACAGTACCCTCATAGAACATCATGTAGTTGTTC

[0583] AGCTCAGCTGGTACTACTGTAGCTGTTCCTGTGTCGTCAACTTTCATGAATTGCATGTCAGCTGAAACCTCAACAGCTTC

[0584] TTCTGCTTTAGCTGAGATTGTAAGAGTATCAATCTTGCCACCTAAGTAGTTACGTAGTAAGCCAGCGCCAGCAATGTTGT

[0585] GTTGAGCTGTGAAAGATGGTAACTCTTGACCAGCCTTGACAGGTTTTAACGTGTGAGTAAAACCAACTGTAGCGTCACC

[0586] TGTTTTGACTTCATTCCCAAGAGCTAAGAACAGCAAGCGTCCATTTTGAAGCGCTCCTTTAATTGAAGCGTCAACTTCTT

[0587] TAGCACCAGTACGTAACATGAAAGCTTCACGTTGTCCAATACCTCTGCGAACGTCTACATTGTTCTTTTCTTCTGGCTCG

[0588] AATGACTCAATAATTGTTGCGTGCTTGAAAATAGTAGCTTTTGTACCTGCTGTAGCTTCCACTCCAATAGTAACCGTATGG

[0589] TCATAGCCTTGTGCTTGTTTTGGCATTAGTTTTTACCTCCTTTGTCTTTTTACTTCTTCAAAGCCATTCTTCTTGAGGTCAGC

[0590] AATCAATTTCTTGTCAGTAACCTCTATCGTACCTTGTGGCTCTACTCTTTCACCAAAAACTGAAAGAGGTATCTCTGTTAT

[0591] ATTTATGAGCTCCATGTTTATCCCTCCATTTTGCCCCTGATAGTAACGACAAGTTTAGCACCTTGAAGGAAATTAGCTTCA

[0592] CCGTTCAAGACTTGACCAAAACTCAGCGTCCTCATCAATAGATAAGTAAGCAGCTGCTCCATTGAGTGTTTTGTCTTCCTC

[0593] AATGATTCTTTCTGCAATTTCTAAAATCTCATAGCATTGCTCCTCCGCTTCTTCACAGTCGAGAATATCAGTGTACACCCA

[0594] CACAAAAAATGTCAAGTTCCTTTCCGAAACTCCAAGTCCTTTTTTGACTTTTCCTCGTTTTGAAAGCTCTATAGCTACAG

[0595] CAGGAAACTGAGCGATATTTTGATAGTGTACTTTGTAGACATCGACCTTTTCAGTTGAAGCATTGAAGCCAGTCACTAAT

[0596] TGATTGAATAAAGCTTCTTTAATCTTTCCGTATAAGTTAGCCATTAGTTAGCCAGCCCCTTTATATATTCCTCAAATACACG

[0597] CTTGATAGCTGCCTCGTCCTTTGAGTCAAAGTACAAGAATTGTCGTTGCGGTATATAACCAAACTTAGTAGCTCCTCCAA

[0598] AGTTATGAATAGCTGCATATGGAAGGTTAGTTCCAAAGTGCAAGCTCTCCCTGCCAACAGTCTTGATAGCTCCACCTGTC

[0599] ACAGAACCTCTAAGAGTACCGCCATCATTTAGTGGCTTTCCTCCAGCTCTATGAGGGTGACGCTTGATAGTACTTGCTGC

[0600] CAGTGGTGTCCAAGCTGCTGCACTAAAACGACGACCAATTGAGCCCTCCATGTAAGTCGCTGACTGAGTAAGTGGCTCT

[0601] TTCATGTTTTTTAACTCGCTATTCATTCTCATGAGGTTAGCTAGTGTTTGCGATATATTGACGGTTATTTCTGTGTTAGTAGC

[0602] CATCAATCACCACTCCGTATCTAGAGTAAACTCTGGGTCTCTGTCGTTCGTAGTAGCGAAACCTGTCTCAGCGCTGTGCT

[0603] TTGGTGGTGTACCAATATTCAAGTTACCTGCTCGTATTTGCGAGATAGTTTCTTGCAATCTAGTCCACCTGTCATTTACTC

[0604] GCTCGTCTAGGTTTGGCTTTTGAGCTGTGTATCTTGCCTCGTGAAACATGCATACTGCAATGTCCGTCGTCAAGTCAGTTA

[0605] TCAATTGAGGAGCTGGGCTGAACGGCACAACAAATAGGTCACTCAGTTTGGCATCTAAATAAGTACTAGCTTTCTGAGC

[0606] AAATACTTCCAAGTCAGCATCCGTTATGCTAGTTCCAAGGTTGGTCATTATCTGTCTCATGAGTGTTGCTGTTGTATAAGC

[0607] CATTCAGTTCGCCTCCTTTACTCTTTTGCTGCTGCCTTCTTAGCAGGAGCTTTTTTAGCTGGTGCTTTGACTTCCTCGGCT

[0608] GCTCCAATAGAAAGCCAGTGAGCTGCTGTCTCCTCGTCCACTTCAATGACTTCCCCAGCAGGTCTGCCACTCAAGCCAA

[0609] CATTTTCCTCTAGTGTTTTAACCTTCATTGTATCGCCTCCATAAAACTAAAGGAGAGCTAAAAGCCCTCCCTAGTGTTACT

[0610] CCAGTATTACGCAATAACGTTCTTTAAGAAGTATACCGCTTTAGGGTCAGACAAGTACGCTTGAGTGTAACGTGTAGCAC

[0611] GTACAATTGTGCGGTCATTTGACTCCTCGTTATATACTTTTGTGCGTAAGCCCTCAGCGTCAGCAATGTCACCGATAACTT

[0612] TCTTTTGAACCATTAAGACATGCTTGTTTTGGAAGTTCTCATCTTGAATGAATGAAAGCCCCATGAAGTCACCAATGTAG

[0613] CCACGTAGTAAAGCTACGTCAGTGTTATTTTGACGGAAAGCGTCACGAATGTCTTTAGTTTTCAACAACATAGCCATAAC

[0614] ACGAGGGTTAACAACTACAGTGTCAAGATTGTATCCGTACTCAGCTGCATCTGTTTTAGCGTCAACTAACTCGCCAATCA

[0615] TTTTGTCAGCTGTAGTCCAACCAGCAGAAGTAATACCAAGACGACCAGAACCGCCAGCATATTTGTTAGTTGCTACGTC

[0616] GTAAGCCATAGTGTCAACCATTTTCTTAACGTTAAGAGCTAACTTACGGTAAGCACGCTCCATGTAACCTGTGTTGCCCC

[0617] ACTTTTGGATTTCCTCAGAGATTACAGCCTCAAGACCGTACTTTTTAATGAACTCTTGTTTCTCAATCTCTGTTAATCCGA

[0618] TACGCTCAAAGTTAGAGCTTTCTCCAACTTCTGGTACTACGTCGTAAGCTTGGCGACCTTTAGCGTCTGCCTCGCCATCT

[0619] TGGTAGTACTTAATAGATAAAGCGTCAACGTTCGTTTGCGTGAATAATTGGTCAGCAACAAACTGGTGACCTACTAAGTC

[0620] ACGAATACGCTTGTCAACGAACTCCTTTTTAAGCATCTGGTGTTGACCTAAGATAATATCTGGCATAATGTAAAACCTCCC

[0621] TTATTTTGAGTTATTTATTAACCTAACGCTACAAGAATAACGTCATTAGCTGCACTTGCAGGTGTAATAGCTAAACCGATT

[0622] TTTTGACTTACTGGGTCTGGAGTAGCTCCAACTTTCAATACAGCCACTTTGTTAGCTCCAGCAGCTTTAATCAATGAACC

[0623] AGCTGGGATAGCCTCACTTGCAGTCATGTATACGAATGGTCGTAAGATAATTACTGTTACTACGTCGCCACGGTCACCTT

[0624] GGTAGCCAACGTTTACACCGTCAACTCCAACAGTCCCAGAGTAAACTACTCCAGCAACCTTCTCAGAGCCAGCAGCAG

[0625] CAGTTGTAACCTCACGAGCTCCAGAAATTTCTACTACGTCGCCAATCTTTAAAGTTGAGCCAGCTTTTACCTTGAAAGTA

[0626] AATGTAGAACCAGCTTGAATGTGGTATTCCACTTTATTACGTGCCATACTTGTCAACCTCCTTGTTTTATGTTATTGTATTAT

[0627] AGGGACGTTCCGTACTTTGCAGCGTGCTCCTCATAAAAAGCATCAAAGTCTTTTTGTTCTTGAGTGCGCTTGTCTTCTGG

[0628] GTCGCCACCTTCTATTTGACCTTTTTCAGATAAGTCAACCGTTTGGTGGTTGCTCATAAATACCTCGAACAATTGAGCTTG

[0629] CTCCTCAGAGAATGAAGCTAGTAAGTTCTTTAATGACTCAGCTTGAGCTGGTACGATTTGCTTAGCTTCTTGGAACTTTT

[0630] CAACTTGAGCTAATACGTTAGCCTCTTTGAAAGAAACCTTTTCAGCTTTTAAGCCTTCCATTTCAGTCTTCATAGCATTGA

[0631] ATTGCTCCATTAACTCACCGAACTGCTCAAGAGTTGGCGCTTTAACTTCTGGGTTTTTGTTTTCTGGCATCTTGTTAGCCT

[0632] CCTTTTCGATAATAAGATACAAAGCCATTCTCACTAAACAACTGAGCTGACTTCACTTGAGGAAAAGCAACCAGTGAAAC

[0633] CTCACGTAGCTTATGAGGTCTTATAGTGTCGTCCTCATCGTTGTACTGAATGTAAAACGAAACTGACAGCTTACCCATGA

[0634] GACCTTTTTCTACACGGTCTTGAGCAAATTCGTCAATGATTCTAACTTTTCCAAGTAGCTTGTCACCCCTTCACTTTTGCCT

[0635] CCTCGAGAAAGCCGACTGTATCACGAACGCTCTCAGAGTGGTCTAGCTGAATAGGTACTCTTTCGTCAGTTGAAAAGTT

[0636] AGTAGCTAAAGTTTCTAAATCTTGAGTTGTTGTACTCTTTGCCTCGGTGTGTTCCTGTGGAGAAAAGCTCAGCCTCTTTAA

[0637] CTATTCCTAGTTGGTCTGGCTCGCCTTCTTCAAAGTTTACAACCTCTGAGAACTTACCCAAGTAGATAAGCCTTGTTTG

[0638] TTTTCTGGCATCTTTCCACCTCCTAAGTAAATTCCGTACTGTCCTTACACTATATATATGACTGGGGTGTTTATCTTTTTAGA

[0639] AAGAGGCGTTACTGGGAGTTAAAAGCTAAAGAGTAAACACCTAGTTCATATATCTTTTGAGGGTTACTAAATAGCTTAAC

[0640] TAATTAGTTCTACCTCATTAGATATTAAAGATTTTAAAGACTTTAAACTATATAGTTATTAACTTAATTAGTAACTAAG

[0641] GGTATAGCTAAATTGATTCAACTAAAAGAAAGAGCTAAAGAGATACTAGTTAGTACTAATAAGTTCACACTCTTAGAGAA

[0642] AACTAAGATGTTATACTCTTTAGTGCTTTGCGTTTACTATGAATTACGAGGCAGAAAAACGTCTAACAAGTTACTGCAAG

[0643] CTGGCTATGACGCTATGTATCTGGCTCACTGGGTTAATCACAAAAGGATTTTCTACAAAGTTACAAAAATAGTTGTTGACC

[0644] TGCTGGTATGACTATGGTAAGATGAGTACATAAATTGATGAGGGAGCTGGTTTGAATGAGTTTAACTTTAAAGTGTGCAA

[0645] CTGATAGCAGAGACGAGTTAGTTTGTGAAGTAGGAGAAATTTCTGGTAACTATATTGTTTCTGGCTATGAAGACGGTGAG

[0646] CACCAGAAGACTGTAACGTTAAACTTGGAAGATGCAAAACGCTTAGAAAACCAAGTGTTTCAGTACCTACATGAGCCTT

[0647] TTGAGCCAAGCGACGATATTGGTCAATATGAGATTTACTGGTGTGCTACTGGCAACAAAGACCGCTTTGAGGCAGAGAA

[0648] TTTGACTGACAGAGACGACAGGGAAGTTCTCGCTCTTGATTTTAATACTTATACGTTTAACCTAAAATAGTGAAACTGCTA

[0649] AACTATTACTGGAAAGACCTACGAAAACTTATCTTGGAGGCGAACTGAATGGAAAAAGCACGAGTGAAAGAAATTCTAG

[0650] AAAACGTTAATGAGCTGGACGCTCGTAAGTCCTCAATGGTGTTTGCTGGAGTGCTTGGCGTGCTAGATTCAATGAGCCA

[0651] GTTCCCAGAAACTAAAGAGGTTGCTTCTCAGTTGTTGCAGCTGACCGACAATTTCACTATTGAGCAAAAATTAAAGGGC

[0652] TGACCTGTTGAGGTAGCCCCTTTTTTTATTCTACGCTTGCTTGTCTAAGCAGGTACTCCATAACGTTATTCTTGCTGTCAA

[0653] ACCCCTTCTGAGGCTTCTCCTCTGGAACAAAGTTATCCTCCCAGTCTTCAAAACGAGTTACTGGAAGCCATACACTTCTG

[0654] CATTGGAAGTGGTTAGGTGGTGAGTAGCTTGCTATCGTAACTTGATTCTCAATGTTCACAATCTTACCGTCAAGGTGCTT

[0655] GCAGAGCGTCGTAGTTCGTCTATCAATAATAGCACTGTACTGTAGAGCCTCTACGAACCCACGGTTTTCTGGTGAGGTAT

[0656] AGCGTGCTAGTCTACCAGCGTTGTACATCTTACTAGTCTCCGTTCTCGCTATTGTTTTAGCGTGCTGCTGAGAGAGCCAA

[0657] GTCTCACTAGTTTCTCTGATAGCTTGCGTCATTTCCTTGATTGTAAAACCTTCCTTGAGCCCCTTAGTAACTATCCCAACTA

[0658] TTTTGTCCCTAACAGTCGCCTCAGTAATGATTGTAATATCTAGAGCGTAATTTTCGAGCCACTGAATAGCTTGAACTGGGT

[0659] AGCGTACTGTGTTGGTCGTTTCAGCTATCAGCGTAATGTCCTCAAAGTGCTCGTCGTCCATGAAGTTATAAAGCTCGTTT

[0660] AGTCTTTTTAGTTCTTCATGAGCACGCAAAATCCCCGTATGAGCTGCTGTTTTAACCAGTTTAGTTATAAGCTTCCTGTAC

[0661] TCCTTGTGAGACGGAATACGAATAGACTGGACCAATACGTCTACTGTGTGGCTGTCGCCCTCCTGTGAGGCTTGTAGGA

[0662] GCTCTGGGAGCATCTTTTTGACAGACTTCATAAGTAGCTGGTGACGTTTTTTATTTAGCCTCTGAGAGTCAGAGAGAAAA

[0663] GCCAGCTCTATCTCATCCATGTCTCGCTCAATGAGCTTCAAGTTGGCTCTTTTTTGTCTCTCCCCGAACGTCACAGTTTTT

[0664] CTAGCAAAGTCAGAGGCTTTTATTTTCCCTCAGAGTTACCGTCAGAGCCTCCAGAATTGTCTGTGTTGCCTCCGTTGTCG

[0665] CCTTCCTTACCGTTCGCCTTCTGAGCGTCTAGCGCTGCTTGTGCAGCCTTCTGAGCTTCCTGTTTAGCTGCTTCAATTTCT

[0666] CGCTTAGTTTCTTCACTCATAGCTGGGAATCCAAGCTCCTCACGTAACCAGTCCTCAGTAGGTGCAATTACTCCAGCAGT

[0667] AATCATTTGAGTGAATACAGTAGCTA

[0668] The BLASTn search function in the NCBI database confirmed previously reported bacteriophages with high similarity to phage DZ1, as shown in Table 1. Whole-genome sequencing and alignment revealed low similarity between this phage and existing phages, indicating it is a novel phage.

[0669] Table 1. Similarity comparison between phage DZ1 and existing phages in NCBI.

[0670]

[0671] Example 4: Preservation of bacteriophage DZ1

[0672] Based on the results of transmission electron microscopy and the genome analysis of phage DZ1, this invention names phage DZ1 as Bacillus cereus bacteriophage DZ1.

[0673] Bacillus cereus bacteriophage DZ1 has been deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong Province, 510070, China. The accession number is GDMCC No. 63387-B1, and the deposit date is April 23, 2023.

[0674] Example 5: Mining of phage DZ1 specific sequences and analysis using high-resolution melting method

[0675] Referring to the gene sequence of Bacillus phage provided in GenBank, it was compared with the whole genome sequence of phage DZ1, and the specific gene gp6 in phage DZ1 was discovered. Based on the DNA sequence of this gene, a BLAST comparison was performed with the phage DNA sequence already available in NCBI. The similarity information of the comparison is shown in Table 2. The nucleotide sequence of the specific gene of phage DZ1 is shown in SEQ ID NO.1, and primers were designed based on this gene sequence.

[0676] Table 2. Comparison of specific sequences of phage DZ1 with existing sequences in NCBI.

[0677]

[0678]

[0679] The primers designed based on gene sequences are as follows:

[0680] F: 5'-AGTCAGCATGTTACAAGCCA-3';

[0681] R: 5'-CGAAATGGTGAAAGCGAAATGG-3'.

[0682] Amplification using this primer pair yields the nucleotide sequence shown in SEQ ID NO.2.

[0683] PCR amplification was performed based on the specific gene gp6 sequence of phage DZ1 (primers: F: 5'-AGTCAGCATGTTACAAGCCA-3'; R: 5'-CGAAATGGTGAAAGCGAAAATGG-3'). The sequence specificity was verified by comparing it with other published Bacillus cereus phage sequences. The results are as follows: Figure 3 As shown in Figure A, this sequence can only be amplified in phage DZ1, and no band is generated after amplification by other phages. Furthermore, based on this characteristic sequence, real-time quantitative PCR amplification was performed, and high-resolution melting (HRM) was used to detect the concentration of phage DZ1 genomic DNA, determining the relationship between the amount of phage DZ1 DNA template and the cycle threshold (Ct). After extracting the genomic DNA of phage DZ1, it was sequentially diluted tenfold, and experiments were conducted at concentrations of 100 ng, 10 ng, 1 ng, 100 pg, 10 pg, and 1 pg, respectively. 2×TB Green Premix Ex Taq II was added sequentially to each well of an eight-tube reaction. 10 μL of the above-mentioned forward and reverse primers, 0.8 μL each, were added to different reaction wells. Then, serially diluted DNA template and 2 μL of sterile water as a blank control were added to each reaction well, and the volume was brought up to 20 μL with sterile water. The reaction was performed on a LightCycler96 using a two-step reaction program. The PCR reaction conditions were: 95℃ pre-denaturation for 30 s, 1 cycle; 95℃ denaturation for 5 s, 58℃ annealing for 30 s, and 72℃ extension for 30 s, for a total of 45 cycles. The HRM curve reaction conditions were: 95℃ denaturation for 10 s, initial melting temperature of 65℃ to start the program, and melting at a rate of 2.2℃ / s to 97℃, with real-time monitoring of fluorescence signal at 5 Readings / ℃ during the process.

[0684] The results of HRM method detection using phage DZ1 genomic DNA concentration are as follows: Figure 3 As shown, when the concentration of genomic DNA of phage DZ1 gradually decreased from 100 ng to 1 pg, the normalized melting peak diagram and melting curve maintained good differences. Figure 3 B, 3C). The linear regression equation between the genomic DNA concentration of bacteriophage DZ1 and the Ct value is y = -3.39063x - 19.8448, with a correlation coefficient of 0.99761. Figure 3 D). In antibacterial agents prepared using phage DZ1, this method can be used to detect the presence of genomic DNA of phage DZ1, with a detection value of 1 pg.

[0685] Example 6: Determination of the one-step growth curve of bacteriophage DZ1

[0686] Preheat TSB (1 mM CaCl2) medium to 37°C. Take 1 mL of the host bacterium *Bacillus cereus* 2432-3 growing in the logarithmic phase, centrifuge at 10000×g for 1 min, and resuspend in 1 mL of TSB (1 mM CaCl2) medium to prepare a bacterial suspension. Mix bacteriophage DZ1 with the bacterial suspension at a multiplicity of infection (MOI) of 0.1, adsorb at 37°C for 5 min, centrifuge at 10000×g for 1 min, filter the supernatant into a new centrifuge tube, and determine the amount of unadsorbed bacteriophage. Simultaneously, resuspend the precipitate in 1 mL of TSB (1 mM CaCl2) medium, take 50 μL of the resuspended culture and transfer it to 50 mL of TSB medium (1 mM CaCl2), and incubate at 37°C with shaking at 200 rpm. Take samples every 5 min to determine the bacteriophage titer, and repeat the experiment three times. A one-step growth curve of phage DZ1 was plotted with sampling time on the x-axis and the logarithm of phage titer on the y-axis. The one-step growth curve ( Figure 4 It can be seen that the incubation period of bacteriophage DZ1 is 10 minutes.

[0687] Example 7: Determination of the killing curve of bacteriophage DZ1

[0688] The host bacterium *Bacillus cereus* 2432-3 was cultured to the stationary phase. 1 mL of the culture was centrifuged at 13000×g for 1 min and resuspended in 1 mL of TSB (containing 1 mM CaCl2) medium to prepare a bacterial suspension. Phages and bacterial suspensions were mixed at multiples of infection (MOI) of 0.1, 1, 10, 100, and 1000, with a control of bacterial suspensions without phages. The mixtures were then added to 96-well plates and incubated with shaking at 37℃ and 200 rpm. Absorbance was measured at 600 nm every 15 min for at least 18 hours, and the experiment was repeated three times. The absorbance (OD) of the 96-well plates was measured with incubation time as the x-axis. 600 Plot the killing curves of bacteriophage DZ1 against host 2432-3 under different MOI conditions on the ordinate. The killing curves ( Figure 5 It can be seen that the phage still has a good scavenging effect on Bacillus cereus 2432-3 under the condition of the minimum MOI of 0.1, and can maintain the growth of Bacillus cereus without phage resistance for 18 hours.

[0689] Example 8: Determination of the temperature stability of bacteriophage DZ1

[0690] Take 1 mL of titer approximately 10 8Phage DZ1 at PFU / mL was incubated at 4℃, 25℃, 37℃, 45℃, 55℃, 65℃, and 75℃ for 1 hour, then appropriately diluted. The titers of the phages treated at different temperatures were determined using the double-layer agar method. The experiments showed that phage DZ1 remained stable at temperatures between 4 and 55℃, its activity decreased at 65℃, and it lost its activity above 65℃. Figure 6 ).

[0691] Example 9: Determination of the acid-base stability of bacteriophage DZ1

[0692] Adjust the pH of TSB medium to different values ​​(1–13) using 1M HCl and 1M NaOH. Place 100 μL of phage DZ1 in mediums with different pH conditions to achieve a final concentration of 10. 8 PFU / mL. After incubation at room temperature for 1 hour, the phage was diluted to an appropriate factor, and the titer of phage DZ1 under different pH conditions was determined using the double-layer agar method. The experiment showed that the phage was stable between pH 5 and 9, its activity decreased at pH 4 and 11, and it lost its activity at pH 1, 3, and 13. Figure 7 ).

[0693] Example 10: Determination of the host spectrum of bacteriophage DZ1

[0694] To determine the potential host range of bacteriophage DZ1, 100 strains of Bacillus cereus were cultured to the logarithmic growth phase, and the lytic ability of bacteriophage DZ1 against these Bacillus cereus strains was then determined using a double-layer agar spotting method. The results are shown in Table 3.

[0695] Table 3 Host spectrum of phage DZ1

[0696]

[0697]

[0698] Note: "+" indicates clear, transparent plaques; "-" indicates no plaques.

[0699] A total of 100 Bacillus cereus strains were identified, including the original isolated host Bacillus cereus 2432-3. Phage DZ1 was able to lyse a total of 33 Bacillus cereus strains, of which the phages covered 21 different ST types, including the vomit-inducing ST26 type Bacillus cereus.

[0700] Example 11: Determination of the antibacterial activity of bacteriophage DZ1 in food matrix

[0701] To verify the antibacterial activity of bacteriophage DZ1 in food matrices, rice and milk were selected as experimental subjects. A milk sample (50 mL) and a rice matrix (1 g of rice flour dissolved in 50 mL of pure water, mixed thoroughly, and then sterilized) were prepared. Single colonies of Bacillus cereus 2432-3 were picked and cultured overnight in TSB medium. The next day, the culture was transferred to the initial logarithmic growth phase, and the OD value was measured. 600 =0.7~0.8, at which point the bacterial count is approximately 10. 8 CFU / mL, diluted and added to the prepared rice matrix and milk samples to bring the final concentration of Bacillus cereus 2432-3 to 10. 3 CFU / mL, and add 100 μL of phage DZ1 to make the MOI 1000 (corresponding to Figure 8 (MOI=1000); with all other conditions the same, except without phage as a control (corresponding to...). Figure 8 (2432-3). After static incubation at 25℃, samples were taken at 0h, 6h, 12h, 24h, 36h, 48h, and 72h to calculate the viable count of *Bacillus cereus*. In rice substrate, phage DZ1 effectively inhibited the growth of *Bacillus cereus* for up to 36 hours. Figure 8 A), while in milk, no regrowth of Bacillus cereus was detected until 72 hours later. Figure 8 B). Overall, phage DZ1 can effectively inhibit the growth of Bacillus cereus in rice substrate and milk, and has good potential for controlling Bacillus cereus in food.

[0702] SEQ ID NO.1

[0703] ATGATGAAAACTAACGAAATGGTGAAAGCGAAAATGGAGATTATCAATCAATTCGAAAATATTGAAGGCTGCAAAGTTATCCTAAAAGTTGAAGAATATGACGGAGAGTTTGTCATTGAAGGCGACGCTTGGGACG CTGACGGTGAGTATGTAACAGATTTTGAACTTTCTCATAAGTTCACTAACTGGAAACATGCTTCAATGGCTTGTAACATGCTGACTAACAAGTTAGCTAAAGAAGACTTTGACTCAGTAGAATACGCTCGCTAASEQ ID NO.2

[0704] CGAAATGGTGAAAGCGAAAATGGAGATTATCAATCAATTCGAAAATATTGAAGGCTGCAAAGTTATCCTAAAAGTTGAAGAATATGACGGAGAGTTTGTCATTGAAGGCGACGCTTGGGACGCTGACGGTGAGTATGTAACAGATTTTGAACTTTCTCATAAGTTCACTAACTGGAAACATGCTTCAATGGCTTGTAACATGCTGACT

Claims

1. A strain of Bacillus cereus bacteriophage ( Bacillus cereus bacteriophage) DZ1, with accession number: GDMCC No: 63387-B1.

2. The application of Bacillus cereus phage DZ1 as described in claim 1 in the preparation of Bacillus cereus antibacterial agents.

3. The application according to claim 2, characterized in that, The antibacterial effect refers to the inhibition of Bacillus cereus in individuals, food, or the environment.

4. The application according to claim 2 or 3, characterized in that, The antibacterial effect is to inhibit Bacillus cereus in food, which is rice or milk.

5. An antibacterial agent, characterized in that, It contains Bacillus cereus phage DZ1 as described in claim 1 as an active ingredient.

6. A primer for detecting Bacillus cereus phage DZ1 as described in claim 1, characterized in that, include: Forward primer sequence: 5'-AGTCAGCATGTTACAAGCCA-3'; Reverse primer sequence: 5'-CGAAATGGTGAAAGCGAAAATGG-3'.

7. A method for detecting Bacillus cereus phage DZ1 as described in claim 1, characterized in that, The primers described in claim 6 were used to target the molecular target of Bacillus cereus phage DZ1 as described in claim 1. gp6 The molecular target was detected. gp6 The nucleotide sequence is shown in SEQ ID NO.

1.

8. The method according to claim 7, characterized in that, The detection using the primers described in claim 6 includes the following steps: extracting DNA from the sample, and then performing PCR amplification using the primers described in claim 6. If a band size of 208 bp is obtained, then Bacillus cereus phage DZ1 is present; otherwise, it is not present.

9. A qPCR method for quantitative detection of Bacillus cereus phage DZ1 as described in claim 1, characterized in that, Includes the following steps: a. Using the primers described in claim 6, plot a standard curve for Bacillus cereus phage DZ1; b. Perform qPCR amplification on the sample using the primers described in claim 6, obtain the ct value of the sample, and calculate the content of Bacillus cereus phage DZ1 in the sample according to the standard curve.

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