Double-target antibacterial lipopeptide and preparation method thereof

Abstract

The invention discloses a double-target antibacterial lipopeptide and a preparation method thereof, and relates to the technical field of biology, the lipopeptide can be combined with double targets on Gram-positive bacteria or Gram-negative bacteria, and the lipopeptide can be synthesized according to a non-ribosome peptide synthesis rule on the basis of a non-ribosome peptide synthesis gene coded by aquimarina sp.EL43. The method comprises the following steps of: directly predicting a substrate (amino acid or organic acid and the like) recognized by the adenylation structural domain enzyme activity central site by using the composition of 10 conservative position amino acids at the adenylation structural domain enzyme activity central site, deducing a linear polypeptide sequence coded by a gene cluster, and then adding myristic acid fatty acid side chains widely used by lipopeptides, and adopting a polypeptide solid-phase synthesis strategy. The invention provides the novel antibiotic which is safe, effective, not easy to generate drug resistance and relatively low in cost, and the preparation method thereof.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and particularly to a dual-target antibacterial lipopeptide and a preparation method thereof. Background Art

[0002] Antibiotic resistance poses a serious threat to global public health. Microbial natural products are the main source for the development of new antibiotics. However, the discovery of traditional natural products is limited to conventional drug-source microbial resources, and the reproducibility of activity screening methods is high, resulting in a lack of natural products with new skeletons and new targets, which severely restricts the research and development of new drugs. Rare Gram-negative bacteria, such as Aquimarina sp., are an underexplored microbial resource, containing rich natural product biosynthetic gene clusters.

[0003] Aquimarina belongs to the family Flavobacteriaceae of the phylum Bacteroidetes and is a type of bacteria from the rare marine biosphere. In the genomes of sequenced Aquimarina strains, there are rich and undeveloped natural product synthetic gene clusters, which may help them adapt to different ecological niches. However, since the first Aquimarina was discovered in 2005, only three classes of polyketides and one class of non-ribosomal peptides have been identified from this genus through traditional fermentation. It is speculated that the main reason is that most natural product synthetic gene clusters are not expressed under standard experimental conditions. With the improvement of the accuracy of non-ribosomal peptide structure prediction and the progress of modern synthetic organic chemistry, it is currently possible to rapidly obtain the metabolites encoded by gene clusters through the method of accurate structure prediction chemical synthesis (i.e., synthetic-BioinformaticsNatural Products, syn-BNP). Therefore, the present invention focuses on the underexplored non-ribosomal peptide synthetic gene clusters of Aquimarina, and uses the syn-BNP strategy to rapidly obtain new-structured antibacterial peptides, thereby obtaining a pair of open-loop or closed-loop dual-target antibacterial lipopeptides, Aquimaricin L and Aquimaricin C4. Both of them have significant antibacterial activity both in vitro and in mice.

[0004] Therefore, those skilled in the art are committed to developing a new type of antibiotic that is safe, effective, not easily resistant to drugs, safe, effective, and has a lower cost. Summary of the Invention

[0005] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is to develop a new type of antibiotic that is safe, effective, not easily resistant to drugs, and has a low cost.

[0006] To achieve the above object, the present invention provides a dual-target antibacterial lipopeptide, characterized in that the lipopeptide can bind to dual targets on Gram-positive bacteria or Gram-negative bacteria.

[0007] In a preferred embodiment of the present invention, the lipopeptide is obtained by a structure prediction chemical synthesis strategy based on the non-ribosomal peptide synthesis gene cluster encoded by Aquimarina sp. EL43. The non-ribosomal peptide synthesis gene cluster has the accession number No. PQ867817 in GenBank, and the sequence is shown as SEQ ID NO.1.

[0008] In another preferred embodiment of the present invention, the dual targets on the Gram-positive bacteria are cardiolipin (CL) and phosphatidylglycerol (PG), and the dual targets on the Gram-negative bacteria are lipopolysaccharide (LPS) and phosphatidylethanolamine (PE).

[0009] In another preferred embodiment of the present invention, the antibacterial lipopeptide is the linear lipopeptide Aquicidine L, with the chemical formula C 70 H 116 N 16 O 16 , and the chemical structure is shown as follows:

[0010]

[0011] In another preferred embodiment of the present invention, the lipopeptide is the cyclic lipopeptide Aquicidine C4, with the chemical formula C 70 H 114 N 16 O 15 , and the chemical structure is shown as follows:

[0012]

[0013] The present invention also provides a preparation method for the aforementioned dual-target antibacterial lipopeptide. The method is characterized in that, according to the non-ribosomal peptide synthesis rule, that is, by using the amino acid composition at the conserved position of the active center site of the adenylation domain enzyme, the recognized substrate is directly predicted, and then the linear polypeptide sequence encoded by the non-ribosomal peptide synthesis gene cluster encoded by Aquimarina sp. EL43 is deduced. Subsequently, a myristic acid fatty acid side chain is added, and the dual-target antibacterial lipopeptide is obtained by using a polypeptide solid-phase synthesis strategy.

[0014] In a preferred embodiment of the present invention, the preparation method comprises the following steps:

[0015] Step 1: Load amino acids onto 2-chlorotrityl resin and swell in dichloromethane (DCM), then drain the solvent and wash with N,N-dimethylformamide (DMF).

[0016] Step 2: Mix the Fmoc-protected amino acids with 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) in DMF, and couple each amino acid step by step. The coupling reaction is carried out for 1 hour, with occasional stirring during the period, and washed with DMF.

[0017] Step 3: Couple the remaining amino acids.

[0018] Step 4: Complete the final cleavage of the linear or cyclic peptide to remove the N-Fmoc protecting group. Dissolve each peptide in the cleavage mixture (95% trifluoroacetic acid, 2.5% triisopropylsilane, and 2.5% water), and react for 1.5 hours. Subsequently, add a pre-cooled 1:1 mixture of diethyl ether:hexane, and let it stand at -20 °C for 10 minutes to precipitate the linear or cyclic peptide.

[0019] Step 5: Collect the polypeptide precipitate, redissolve it in methanol, and dry it under vacuum conditions.

[0020] In another preferred embodiment of the present invention, when preparing the linear lipopeptide marinobacterin L described in claim 4, the synthesis starts from the last amino acid. Step 3 specifically includes: The Fmoc deprotection is carried out using a 20% piperidine in DMF solution for 7 minutes, repeated twice. After the resin is washed with DMF, it is coupled with the next N-Fmoc-protected amino acid.

[0021] In another preferred embodiment of the present invention, when preparing the cyclic lipopeptide marinobacterin C4 described in claim 5, the synthesis starts from the fifth amino acid.

[0022] In another preferred embodiment of the present invention, Step 3 specifically includes: Mix the resin with benzoyl chloride, DIPEA, the last amino acid of the polypeptide, and 4-dimethylaminopyridine (DMAP) in DCM, and slowly rotate for 72 hours. After the formation of the ester bond, couple the remaining amino acids according to the method of Step 2.

[0023] The resin is removed by treating with 20% hexafluoroisopropanol (HFIP) in DCM for 2 hours.

[0024] Subsequently, the soluble peptide is cyclized using (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PYAOP) and DIPEA in DMF, and extracted with DCM and H2O containing 1% formic acid. The cyclized polypeptide is dissolved in methanol and dried under vacuum overnight.

[0025] Technical effects

[0026] 1. The present invention provides a pair of antibiotics with novel structures and dual targets that have antibacterial activity in mice. Among them, maribactericin L mainly targets lipopolysaccharide and phosphatidylethanolamine in the cell membrane of Gram-negative bacteria, while maribactericin C4 mainly targets cardiolipin and phosphatidylglycerol in the cell membrane of Gram-positive bacteria. By using the structure prediction chemical synthesis strategy, the non-ribosomal peptide synthesis gene cluster of Maribacter was rapidly decoded, and thus new structure antibacterial peptides were obtained. Two lead compounds with novel structures and unique targets were provided for the treatment of infections caused by drug-resistant pathogenic bacteria.

[0027] 2. Since maribactericin L and maribactericin C4 act on the bacterial cell membrane and have dual targets, it is not easy to produce drug resistance. Using a 21-day continuous drug resistance induction experiment, directly observe whether maribactericin L and maribactericin C4 are prone to produce drug resistance. Two lead compounds that are not easy to produce drug resistance were provided for the treatment of infections caused by drug-resistant pathogenic bacteria. Compared with the approved antibacterial lipopeptides daptomycin and polymyxin in the past, maribactericin L and maribactericin C4 have basically equivalent antibacterial activities and are not easy to produce drug resistance, and have good substitutability.

[0028] 3. The linear peptide maribactericin L has better activity against Gram-negative bacteria, while maribactericin C4 has better activity against Gram-positive pathogenic bacteria, and the activity switches when it is open-loop and closed-loop. Chemically directly synthesize the linear and cyclic peptides that may be encoded by the maribactericin synthesis gene cluster, and measure and compare the antibacterial activities. A pair of lead compounds with interchangeable antibacterial activities were provided for the treatment of infections caused by drug-resistant pathogenic bacteria, and a new idea was provided for the study of the activity diversity of other antibacterial peptides.

[0029] 4. The present invention obtained two lead compounds acting on drug-resistant pathogenic bacterial infections through chemical synthesis, namely the antibacterial lipopeptides maribactericin L and maribactericin C4 with new structures and dual targets, and conducted detailed pharmacological studies on them, including in vitro antibacterial activity, cytotoxicity and hemolytic activity, acute toxicity and antibacterial activity in mice, and analysis of antibacterial mechanisms, etc. Before entering clinical research, systematic preclinical studies, including efficacy, pharmacokinetics and toxicology tests, etc., are still required.

[0030] 5. Maribactericin L and maribactericin C4 are obtained by the conventional solid-phase synthesis method of polypeptides. The large-scale production price is 3000 - 5000 yuan per gram, the cost is relatively low, and the production implementation has high feasibility.

[0031] In summary, the two antibacterial lipopeptides maribactericin L and maribactericin C4 obtained by the present invention have good industrial application prospects.

[0032] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the drawings to fully understand the purpose, features and effects of the present invention. Description of the Drawings

[0033] Figure 1 It is a schematic diagram of the chemical structure of known natural products (including a class of non-ribosomal peptides and three classes of polyketides) from the genus Aquimarina in a preferred embodiment of the present invention;

[0034] Figure 2 It is a schematic diagram of the chemical structure of a dual-target antibacterial lipopeptide in a preferred embodiment of the present invention;

[0035] Figure 3 It is a schematic diagram of the analysis results of the antibacterial mechanisms of aquimaricin L and aquimaricin C4 in a preferred embodiment of the present invention;

[0036] Figure 4 It is a schematic diagram of the results of the cytotoxicity, hemolytic activity and acute toxicity in mice of aquimaricin L and aquimaricin C4 in a preferred embodiment of the present invention;

[0037] Figure 5 It is a schematic diagram of the test results of the antibacterial activity in vivo of aquimaricin L and aquimaricin C4 in mice in a preferred embodiment of the present invention. Detailed implementation manners

[0038] The following introduces multiple preferred embodiments of the present invention with reference to the accompanying drawings of the specification, making its technical content clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0039] Three classes of polyketones and one class of non-ribosomal peptides were identified from the genus Aquimarina through traditional fermentation, and their chemical structures are as Figure 1 shown.

[0040] The present invention adopts the strategy of structure prediction chemical synthesis (i.e., synthetic-Bioinformatic Natural Products, syn-BNP). Based on the non-ribosomal peptide synthesis gene cluster (Aquicidine biosynthetic gene cluster, GenBank No. PQ867817, sequence SEQ ID NO.1) encoded by Aquimarina sp. EL43, two corresponding lipopeptides were directly predicted, including linear lipopeptide aquimaricin L and cyclic lipopeptide aquimaricin C4 ( Figure 2)。Specifically, according to the non-ribosomal peptide synthesis rule, that is, by using the composition of amino acids at 10 conserved positions in the active center site of the adenylation domain enzyme, the substrates (amino acids or organic acids, etc.) it recognizes are directly predicted, and thus the linear polypeptide sequence encoded by the gene cluster is deduced. Subsequently, the widely used myristic acid fatty acid side chain is added, and the linear lipopeptide maribactin L and the cyclic lipopeptide maribactin C4 are obtained by using the solid-phase peptide synthesis strategy.

[0041] Example 1: Preparation of Maribactin L

[0042] 1) In the process of maribactin L, start from the last amino acid. First, load the amino acid onto 2-chlorotrityl chloride resin (0.3 g, 0.552 mmol / g), and swell it in dichloromethane (DCM) for 20 minutes, then drain and wash with N,N-dimethylformamide (DMF) (3 mL, repeated 3 times). Mix the Fmoc-protected amino acid (2 equivalents relative to the resin loading), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (2 equivalents), and N,N-diisopropylethylamine (DIPEA) (2 equivalents) in DMF (5 mL), and gradually couple each amino acid. The coupling reaction is carried out for 1 hour, with occasional stirring during the period, and washed with DMF (3 mL, repeated 3 times). Fmoc deprotection is carried out using a 20% piperidine in DMF solution (3 mL) for 7 minutes, repeated twice. After the resin is washed with DMF (3 mL, repeated 5 times), it is coupled with the next N-Fmoc-protected amino acid;

[0043] 2) After the final cleavage to remove the N-Fmoc protecting group of the linear peptide is completed, dissolve the peptide in 3 mL of cleavage mixture (95% trifluoroacetic acid, 2.5% triisopropylsilane, and 2.5% water), and react for 1.5 hours. Subsequently, add a pre-cooled ether:hexane (1:1) mixture, and let it stand at -20 °C for 10 minutes to precipitate the linear peptide. Finally, collect the polypeptide precipitate, redissolve it in 5 mL of methanol, and dry it under vacuum conditions.

[0044] Example 2: Preparation of Maribactin C4

[0045] 1) The synthesis of marinobacterin C4 mediated by ester bond cyclization starts from the fifth amino acid (AA). First, the amino acid was loaded onto 2-chlorotrityl resin (0.3 g, loading capacity of 0.552 mmol / g) and swollen in DCM for 20 minutes. Subsequently, the solvent was drained and the resin was washed with DMF (3 mL, 3 times). Then, the Fmoc-protected amino acid (2 equivalents, relative to the resin loading), HATU (2 equivalents), and DIPEA (2 equivalents) were mixed in DMF (5 mL), and each amino acid was gradually coupled. The coupling reaction was carried out for 1 hour with occasional stirring, and the resin was washed with DMF (3 mL, 3 times). Then, the resin was mixed with benzoyl chloride (20 equivalents), DIPEA (40 equivalents), the last amino acid of the polypeptide (20 equivalents), and 4-dimethylaminopyridine (DMAP) (0.8 equivalent) in DCM (10 mL) and slowly rotated for 72 hours. After the formation of the ester bond, the remaining amino acids were coupled as described above. The resin was treated with 20% hexafluoroisopropanol (HFIP) in DCM for 2 hours to remove it. Subsequently, the soluble peptide was cyclized in DMF using (7-azabenzotriazol-1-oxy)tripyrrolidinophosphonium hexafluorophosphate (PYAOP) (8 equivalents) and DIPEA (30 equivalents), and extracted with DCM and H 2 O containing 1% formic acid. The cyclized polypeptide was dissolved in methanol and dried overnight under vacuum;

[0046] 2) After the final cleavage of the cyclic peptide to remove the N-Fmoc protecting group, the peptide was dissolved in 3 mL of cleavage mixture (95% trifluoroacetic acid, 2.5% triisopropylsilane, and 2.5% water) and reacted for 1.5 hours. Subsequently, pre-cooled ether:hexane (1:1) mixture was added and allowed to stand at -20 °C for 10 minutes to precipitate the cyclic peptide. Finally, the polypeptide precipitate was collected, redissolved in 5 mL of methanol, and dried under vacuum.

[0047] Subsequently, a variety of Gram-negative bacteria, Gram-positive bacteria, and pathogenic fungi were selected and their activities were tested (Table 1).

[0048] Table 1 Antibacterial activities of marinobacterin L and marinobacterin C4

[0049]

[0050] Note: All the above strains were purchased from the China Center for Type Culture Collection.

[0051] The results showed that marinocine L had certain antibacterial activities against both Gram-negative and Gram-positive bacteria, especially significant activities against Gram-negative pathogenic bacteria, with the MIC value reaching 4 μg / mL. Meanwhile, marinocine C4 also had certain inhibitory activities against a series of different Gram-negative and Gram-positive bacteria, especially significant activities against a series of different Gram-positive pathogenic bacteria, with the MIC value reaching 2 - 4 μg / mL.

[0052] In the preferred embodiment, the activities of linear lipopeptide marinocine L and cyclic lipopeptide marinocine C4 against Gram-negative and Gram-positive bacteria were interchanged. The linear state had significant activities against Gram-negative bacteria but poor activities against Gram-positive bacteria, while the cyclic state had significant activities against Gram-positive bacteria but poor activities against Gram-negative bacteria. The structure prediction chemical synthesis synBNP strategy could obtain both linear and cyclic peptides simultaneously, and then compare and evaluate their activity differences.

[0053] In other embodiments of the present invention, the antibacterial mechanisms of marinocine L and marinocine C4 were analyzed. As Figure 3 shown in a of, marinocine C4, rather than marinocine L, could cause rapid lysis of Staphylococcus aureus cells. Such a phenomenon was not caused by the accumulation of cell wall precursors, but by the depolarization of the cell membrane caused by marinocine C4, indicating that its action target was the components of the cell membrane ( Figure 3 b and c of). The results of in vitro small molecule addition experiments and isothermal titration binding experiments showed that marinocine C4 had a stronger binding ability to cardiolipin (CL) and phosphatidylglycerol (PG) compared with marinocine L, and the cell membrane of Gram-positive bacteria (such as Staphylococcus aureus) was mainly composed of CL and PG, thus explaining the better antibacterial activity of marinocine C4 against Gram-positive bacteria than marinocine L. On the other hand, marinocine L had a stronger binding ability to lipopolysaccharide (LPS) and phosphatidylethanolamine (PE) compared with marinocine C4, and the cell membrane of Gram-negative bacteria (such as Escherichia coli) was mainly composed of LPS and PE, thus explaining the better known activity of marinocine C4 against Gram-negative bacteria than marinocine L ( Figure 3 d - h of). Through the above genetic and biochemical experiments, the present invention not only analyzed the antibacterial activity interchange mechanism of linear lipopeptide marinocine L and cyclic lipopeptide marinocine C4, but also found that both of them had dual targets.

[0054] In a preferred embodiment of the present invention, a drug-resistant mutant induction experiment was simultaneously carried out to observe the tolerance of pathogenic bacteria to marinobactin L or marinobactin C4. The results showed that in the case of continuous induction for 21 days, marinobactin C4 with dual targets (CL and PG), compared with daptomycin with a single target (target: PG) or telomycin with a single target (target: CL), when acting on methicillin-resistant Staphylococcus aureus USA300, the pathogenic bacteria were not likely to develop drug resistance. At the same time, marinobactin C4 with dual targets (LPS and PE), compared with colistin with a single target (target: LPS), when acting on meropenem-resistant Klebsiella pneumoniae CRKP5452, the pathogenic bacteria were not likely to develop drug resistance. Figure 3 i). In the context of global antibiotic resistance, due to the dual-target action mechanism of marinobactin L and marinobactin C4, different clinically drug-resistant pathogenic bacteria are not likely to develop drug resistance to them, so they have good prospects for clinical development.

[0055] Among them Figure 3 Each figure is as follows: a) The cell lysis results of marinobactin L and C4 on Staphylococcus aureus USA300; b) The results of the accumulation of cell wall precursors of marinobactin L and C4 on Staphylococcus aureus USA300; c) The results of the depolarization of the cell membrane of marinobactin L and C4 on Staphylococcus aureus USA300; d) The change multiples of the MIC values of marinobactin L and C4 on Staphylococcus aureus USA300 when adding CL or PG; e) The change multiples of the MIC values of marinobactin L and C4 on Escherichia coli DH5a when adding LPS or PE; f) The in vitro binding experiment results of marinobactin L, marinobactin C4 and CL or PG; g) The in vitro binding experiment results of marinobactin L, marinobactin C4 and LPS or PE; h) The schematic diagram of the interchange of the antibacterial mechanism and activity of marinobactin L and C4; i) The results of the 21-day continuous drug resistance test of marinobactin L and C4.

[0056] In a preferred embodiment of the present invention, the toxicity of marinobactin L and marinobactin C4 to human-derived cells HeLa was tested, and it was found that neither of them had obvious cytotoxicity. Figure 4 At the same time, the hemolytic activity of marinobactin L and marinobactin C4 was tested using sterile defibrinated sheep blood, and it was found that neither of them had obvious hemolytic activity. Figure 4of b). On this basis, the present invention carried out acute toxicity and antibacterial activity tests in mice. The experimental mouse strain was 6-week-old ICR female mice. The results showed that when Bacillomycin L and Bacillomycin C4 were intraperitoneally injected at a high concentration of 250 mg / kg and continuously observed for 1-5 days, all the mice survived, indicating that Bacillomycin L or Bacillomycin C4 had no obvious acute toxicity( Figure 4 of c). Subsequently, a septic peritonitis model was used to determine the in vivo antibacterial activity of Bacillomycin L and Bacillomycin C4. First, Meropenem-resistant Klebsiella pneumoniae CRKP5452 and Meropenem-resistant Acinetobacter baumannii 1104008 were used. After continuously injecting Bacillomycin L four times at 12-hour intervals, the mice were continuously observed for 1-5 days to see if they survived( Figure 5 of a). The results showed that Bacillomycin L had a significant antibacterial effect on the two pathogenic bacteria when intraperitoneally injected at a concentration of 25 mg / kg, and the survival rate of the mice reached 80%. When the concentration was 50 mg / kg, the survival rate of the mice reached 100%. Secondly, Vancomycin-resistant Enterococcus faecium 35682 and Methicillin-resistant Staphylococcus aureus USA300 were used. After a single epidermal injection of Bacillomycin C4, the mice were continuously observed for 1-5 days to see if they survived( Figure 5 of b). The results showed that Bacillomycin C4 had a significant antibacterial effect on the two pathogenic bacteria at concentrations of 100 mg / kg and 150 mg / kg, and the survival rate of the mice reached 100% in both cases. These research results indicate that Bacillomycin L and Bacillomycin C4 have no obvious acute toxicity and can effectively inhibit the growth of a variety of drug-resistant Gram-negative and positive pathogenic bacteria in mice respectively, providing two lead compounds with new structures for solving the global antibiotic resistance problem.

[0057] Among them Figure 4 and Figure 5 are respectively

[0058] Figure 4 : a) The cytotoxicity of Bacillomycin L and C4 to human-derived HeLa cells; b) The hemolytic activity of Bacillomycin L and C4 to sterile defibrinated sheep blood; c) The acute toxicity of Bacillomycin L and C4 in mice.

[0059] Figure 5 : a) Survival results of mice treated with marinobactin L against the growth of two drug-resistant Gram-negative pathogenic bacteria in mice; b) Survival results of mice treated with marinobactin C4 against the growth of two drug-resistant Gram-positive pathogenic bacteria in mice.

[0060] The preferred specific embodiments of the present invention have been described in detail above. It should be understood that those of ordinary skill in the art can make many modifications and variations based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by those skilled in the art in the technical field of the present invention based on the concept of the present invention through logical analysis, reasoning, or limited experiments on the basis of the prior art should fall within the protection scope determined by the claims.

[0061] Sequence Listing

[0062] >GenBank No.PQ867817

[0063] SEQ ID NO.1

[0064] TTCATAAAAATTCTTACTAAAAAGGAGTTGTGAGTCATGTGGCGAAGCTATATCCTTAATTAAATAAACCTATTATACTCTTAAAACCAACAATATGT

[0065] GTATTAGTAAACCTAACCGATGGTACGTGCTATATCTAAAAGCACACCATAAAAGAAAAATAGAAAGCCTGTTGATGGAAAATCAAATAATGGCAT

[0066] TTTAAAATAGATAGTGTTTCTTAATCTTGTACTTAGAAGGTATTCTTCTCCCTCCTCTATTATTTTCTCTTAAATTTCTCGTTGAAGGGTTAATTCCCT

[0067] GTGACTTACCTCGAAAACATATCATGTTTTCGGAAGTATACTTCGTGAGTTTGTCTCAAGATTATTGATTAACACTAAATCACAACTTTTTTAATAGT

[0068] AAAAAAGAAATTTCAAGTAAAAAATACAATCAAATTTTAAGAGTACGTATATCCTGTTATTCAGTCAAAACAGGTCATTTTCGATTGTTAAATAACT

[0069] AAAAAATAAAATATGTTTGAAGAGTTTCACTTATCACAAGAAGCCAAAATAGTATCATCAGAACTTCCTGGTAAACGATCAAAAGTGCTGTTGGAT

[0070] ATCCAACAAGAGTATGAAGGAAGTATTGTCTCTTACCCCAGAAAAATGCCAATCGCAATTAAAAAAGCAAAAGGAAGCATTATTGAAGATGTAGA

[0071] TGGAAATCTGTTTATCGATTTTTTCAGCGGATGCGGAGTACTTAATGTAGGGCATTGTAATGATTTTGTTTTAGAATATGTAAGAAATCAGCAATCAG

[0072] AACTGATTCATGCTCTTGATTTCCCTACAGAGAATAAAATGCTGTTAATTCAAAAAATACTGGATCATTTACCCAAAGATATAAGACGTGATTATAA

[0073] GGTGAGTTTTGGTGGGCCGACAGGCTCGGATGCAGTAGAAGCCGCTATCAAATTAGCTAAAATAAAAACCGGAAGAGATGGTATTATAGCTTTTA

[0074] CCGGAGGGTATCACGGAATGACCTCTGGGGCTTTAGCAGTGACTTCGGATACATATTTTAGAAATAACATAACCTCTTTGATGCCGAATGTACATTT

[0075] TGCTCCTTTCAGTTATCCTTACAGAAGTGTATCTGATAAAGATAATGAGATAGATTATTTTAAAAATATATTAACGAATTCGCATTCGGGAATACCCA

[0076] AACCAGCAGCAATTCTAGTCGAGCCTATTCAAGGTGAAGGAGGAAATATAGTACCCGAGGATGGTTACCTGGAAGAACTGGTTAGGATAGCCCGA

[0077] GACAATGACGTATTAGTCATATTTGATGAAATTCAAAGTGGCTTTTTTAGAACAGGAAGCTTTTTAGAATTTATGAACACAAAGGCGGTACCTGATA

[0078] TAATTACTTTTTCTAAAGGATTCGGAGGAATAGGTTTTCCAATTTCGGGATTAATCTATAGAAGAGAAATAGAAGCCTGGGGACCAGCTGCACATAT

[0079] AGGGACTTTTAGAGCCAATCAGGTAAGTATTGCTGCATCCCGAGGGGCACTGGAGTTTATAGAGAAGTATAACGTACAAGAGTACACCAAAAAGA

[0080] TAGGAGAGTATTTTATTAAAAAGCTGTACAGGTTACAAGAAGATTTCGATTTTATTGGAGATGTAAGAGGAAGAGGAATGATGATCGGAGTAGAAA

[0081] TAGTTAAAGATGTTTTAACCAAAGAACCATTTCCAGAATTTGTAACAATATTTCGAGATGAATGCTTTAAAAGAGGACTCTTGTTTGAAGTTGGTG

[0082] GGCACTACAATAACGTAATACGATTTGTTCCTCCGTTAATTACCACCACCAAAATTGTAGATGCTGCCATAGAAATTTTGGAAGAAGCAATTACAAT

[0083] CTGTACCAAGAAAAATATGGAACACTCTATAGGATAATCAAACATTGAATCTCTCAAAAAACCAAAAATCAAAAACTAATGCTATCTAAAAGCCTG

[0084] TTGATCGAAAAAGGAAATGTAAATAATCAAACGATAATAGACCTATTCAAAACACATGTAAAAACAAAAGAAAACAAAGTCGTTTATAGATTTCTT

[0085] GAAAATGGCGAGGAAGAAACAGATGTAAGAACTTTTGGACAATTGCATAGTAATGCCATGACAATAGCTTCAAGAATCCTTACACTGGTTTCACC

[0086] TGGAGAGCGAGTACTTCTTCTTTATCCATCTGGATTAAATTTTACAGATGCTTTTTTTGGTTGTTTAATGGCCGGAGTTATTGCAGTTCCGGCATTTC

[0087] CTCCCACCGGTAAAAGACGAATAGGGAGATTAGAAAATATAGCTACAGATTGCGATATAAAACTAATTCTTACCGAAGAAAAAGTACATGCCAAAT

[0088] GTAAAAGTTGGTTTGAGCATGAGATTTTTTCAGAGGTTTTATGGTCTGTAACAGATGAACCACATGTAGCTCATAAACCTGTAGAATTACCTTCTGT

[0089] AGATTCAGAAGATGTTGCCTTTTTACAGTATACTTCGGGTTCTACCGGAAATCCTAAAGGGGTTATGGTTTCTCATAAAAATATTATGCATAATACCG

[0090] AATTGATTCGGCAATGTCTTAACCTTAAAAAAGATTTTATTGGAGTAAGCTGGTTACCGATTTATCATGATATGGGGTTGATCGGCAACATTTTAGA

[0091] AGCTGTAGCAATGGGGATAGAAATGATTATTCTTCCTCCACAGGCATTTATACAAAAACCTATTCGGTGGTTTCAGGTTATCTCGAAATACAAAGCA

[0092] AATTATAGCGGAGGCCCCAATTTTGCTTTTGATTTATGTATTAATCAAATCAAAGATGATGATCTGGATAGCATAGATTTAAGCTCTTTATCTACAATG

[0093] TATAATGGAGCCGAACCTATACGTGCTCAAACTATTAAAGATTTTTTAGTCGCTTTTGAAAAAACAGGAATTACCAAAGAGGCTATTTGTCCTTGTT

[0094] ATGGTATGGCAGAGACTACTCTTGCAGTAACTTTTAGCAGATTAGATAAAGAAACTTCCTTTTTAGAACTTGATATAAATGAGTTCGAAAAAGGTA

[0095] AAATCTCTTCAGGTTCCCAACAGAAATCAAGTTCGAATGAAGTAAAAGTATTAGTTGGAAATGGCCCAATATTGCAGGATATGCGAATTGCTATTG

[0096] TTAACCCAAATACCAAAAGAAACTGCTCAGAAGAAGAAATAGGAGAAATATGGGTTAAAGGACCTAGTGTAGCTATAGGGTATTGGGGGAAACC

[0097] CGATCTTTCTGAGGAGATTTTTCAGGCCGAAATACTAGACGAAAAAGGACAACCTCTGGAGGAAAATTGTCAATACCTTAGAACAGGAGATATGG

[0098] GGTTTATGAATAATAATGAACTCTATGTTTCTGGAAGATTAAAAGAAATGATGATCATTAATGGAGTGAATCATTATCCCCAGGATATTGAAAAAAC

[0099] AGTGCAGGAATCACATACCGATCTACAAGTTAATGCAGGTGCTGCTTTTTCTATTGCAATAGGAAGCGAAAACAAATTGGTTATTGCACAAGAAAT

[0100] AAAAAGGACAAGTTTAAGAAGTTATGATTTTGATTCAATTAGGAAAACCATTACAAAATCTATTTTAGAAACCCATGACCTTTCTGTATTTGCCATA

[0101] GTATTACTAACTCCTGGTGGTGTTTCTAAAACTACCAGCGGAAAAATCCAACGATTAGTTATAAAAAAAGCATATGAGTCTAATGCTCTAGAAAAA

[0102] GTAAAAGAAACATGGATTGTCTCGGGAAGCAAAAGGGGTATTAAAAAAGATAAAAGCAATAGTATTGTTTCAGAAAACTTAGAAGATAAAAAAA

[0103] CATTAGATCCGAAGATAATTAATAGGATTATGATGATTTTGGGTGAAGAAACCGAATTTCCCTTGTATCAAATTCAACCCAATACTTCTTTCTTTGAA

[0104] TTAGGAATATCCTCGATACAATCTATTCGAATAGCAGGAAAAATATCTGAGTATTTTGAGATAGATGTACCTTCTTCATTGTTATTAGAATATCCTTCG

[0105] ATTGAAGAGTGTGCAGACTTTTTGTCGACGGAATTAGAAAATGCAAAAAATAAGAACCAGGCCAAAGATACCATCGTAAAAATTAAAGATGAAC

[0106] TGATTAAAATCCCATTGACCTATGCGCAAGAAAGTTTATGGTTGATAGACCAAACATATGGTAGTGTTGATTATCACCTGCATAGAGCGATTAACTT

[0107] ATCTGATTTTCCTGATATTTCTTGTATAGAAGAAGCTTTACAAATCTTAGTCGTTAGGCATGAGGCATTTCGATCTGTAGTTAAGATAGATGAAAACG

[0108] GAGCTTTATATCAAGAGGTTCTAGAGGATTTTTCATTTAGAGTTACGCATCAAATATTAGAGAAGCATCAAAATTTAGAACAAGAGATTGAAGCAA

[0109] ATACCAATATTCCTTTTGACTTATCATCTGATCTTAAAATTCGAGTTACGCTATTCGAAACTGAAAAAAAAGAGTATGTTCTCTTGTTTGTATTGCAT

[0110] CATATTGCTGCAGACGGGCTATCCGAACCTATATTTTTCTCAGAGTTTGTCTCATGTTATACGAATCTTCTTAAAGGGCAACACCCTAATTTGGAAA

[0111] AATCAACGGTTAGATACATAGATTATGCAGCCTGGCAACGTAATCCACATCAGGAGAAAGTTTTAGAAGAAAAGCTGGATTATTGGATAACACAAT

[0112] TACAAGGTATTGTCCAGATCAATGTACCATATGATTTTTCAGTCGTTGGTCAGCACGATAAAAAAGGAGAGAAAATTTCGCTTGTCCTGGGTAAAG

[0113] AATTAAAAAATAATTTACTTCAATTCTCCAAATCAGAAGGCGTTACGTTGTATATGACAATGCTAGCGGCATTCAAAGTCTTACTTTATAAGTACAG

[0114] TGGTCAGTCTGATATTTGTATAGGAACTCCTATTGCAAACCGAACACAAACTGAGGTTTTTAATTTGGTCGGAATGTTTGTCAATACGTTGGCATTA

[0115] CGAAGTGATTTAGGAGGTAACCCAGATTTTAGAGAGTTTTTACTAGAAGTAAAAAATACTACCCTAAGTGCTTATTCACATCAATCTGTTCCTTTCG

[0116] AAAAGATTGTAGAACGATTATCTCCTAATAGGGAATTAGGCAAAAATCCATTATTTCAGATTATGTTTGATTTTCAGGACCATTTTGATCTTGATGGA

[0117] TTTCAATTAGGAGAAATAACTCTTTCTGAGCACCTGATAAAACATACTATATCTCAATTTGATTTCACATTAGAAGTGCGAGATATTCCTGAAGCAC

[0118] TTAATTTATCAATAGAATACTCAACCTCTTTATTTAAGGAGTCTACGGTAAGGCGCTTATTATCACATTATGAAGAACTGTTAAAGAGTATTGTCAAT

[0119] AATGTTTCTTCTCCTATAGATTCTTTATCGATGCTGCCAAAGCAGGAAGAAGAATTAATTTTGGGTCGAAGAGTTACACCAGCAGGACTAAGTTTC

[0120] AATCCAGGAGCAGTTGATTTGGGTAATGAATCTCCGATCAATGTTCGTTTTGAATCCATAGTAGCAACTGATCCTGGTGCTGTAGCATTAATACACA

[0121] ATGAGAAGATTTGGAGTTATAGCGACCTTAATTCGTATTCAAACCAGATTGCTCACAGTTTGTTGTCGTTAGATCTTTCGCCTTCTTCTATAGTTGG

[0122] AGTGTATATGGATCGTAGTGTTGAATTTGTAGGTTGTTTACTGGGTATTTTCAAATCGGGTCATGTGTATACTCCTTTGGATACCAATAACCCACCGT

[0123] CTCGTATAGGTTCGATGTTATCGGATAATCCATTTTCTGTATTAATTACAACAAGTTCTTTATTGTCAGAATTAGGAGATTTGAAAGATATCATAGTTT

[0124] TGGTAATTGATGAATTGCCTGATGATTCTATAGAAGATAAGGTTGTTTATGATAGCAATATGATTGCAGGTAAACCTAAGGATAATCCTGAGAACAG

[0125] GAATTTAATGGATAGCTGGGCGTATATTTTATATACTTCGGGATCTACGGGAGCTCCCAAAGGGGCTATTACACGTCATGATGGGGCGATGAACCAT

[0126] ATTTTGGCAGAGTATGCTGTTATGGATTTAGCAGATGGGTTCCGATTCTTACAAAGTGCAGGTATTGGTTCAGATATTTCAGTATGGCAGATTTTAG

[0127] GTCCTTTACTCAAGGGAGGAGTAAGTGTAATCGTCGACAAGTATGAGTTATTAGAGTATGGCAAGTTGTTGGATACCTTGATTCGAAGAAGAGTTA

[0128] CTCTTATCGAGTTTGTACCAACCTATATGTGGGGATTATTGTCGTATATAAAAGAACAGGAAGAAGCGGTTATTTTGAAAGATCTATCAAGCATTAT

[0129] GCTTGTGGGCGAGTCTATTCCTGTTGCGATGGTTAATGAACTGAAACGTTTGTATCCCGAGGTAAGGTTATGGAATGCTTATGGTCCTTGCGAGGC

[0130] ATCCGATGATGTTATCCAGTATGAGATCAAAGACTTTTTAGAGGAGACCCAGGTACGAGTTCCTATCGGTCGTGTTATTCCTAATATGAACACTGTT

[0131] ATTTTGGATAAATCGGGAGGGTTGTGTCCGATAGGAGTTATAGGAGAGATAGGTGTTAGTGGAGTAGGAGTAGGAGCAGGATATTTAGGATTACCA

[0132] GATCGTACTTCAGAGAGTTTTATAGAGAACCCCTTCAAAGATTTATTAGGCGATGTTTTGTATAAGACAGGAGACTTAGGTCGTTGGCTTGAAGAT

[0133] GGGAACATTGAATTTATAGGAAGGGAGGACCACCAGGTTAAGATTCGAGGACATCGTGTAGAGTTAGAGGATATAGCTTCAGCTTTACGTAAGGA

[0134] TGCTTATGTTGAGGATTGTCATGTTTTGGTTCACAGGACAGGAGGTCAGGATTTAGTGATTTGCTTTGTTGTATTAAGTTCTCAAGGCTTATTGTAT

[0135] GCTTCAGATTCTTCAATTACAGAAGTTTTCCATTTGTTATGCAAGGATGAGTTACCGAGTTATATGCATCCTTCTCATTATCATATTGTAGATGAGTTT

[0136] CCATCAAATTTGAGTGATAAAGTAGATGGCAAGGCTTTGTTAGATATTTTCTTATCAGATTTTGATGATGGAGTTAGTCTTTCTCAACATGTTTATGA

[0137] GGCTCCACGTAATGAAATAGAGGAACACCTGGTTGATATCTGGCAGGAATTGTTGCATATAGACAAAATCGGAGTTTATGATAATTTCTTTGAGTTA

[0138] GGAGGTCATTCATTATTAGCGACACAACTGGTGTCTATGATCCGTAAGCAACTAAACATAGAAGTAGCAATAAAAGATATATTTATACATGCCCGGT

[0139] TGTCTGATCTAGCAGTTCACATATCAGAACAGACATCAGGAGTTTTATTACCGGCAATTACTCCTCAGGAGAGATCAGAACATATCCCTTTATCCTA

[0140] TAGCCAGGAACGTTTATGGTTTTTGGATGAATTAGAAGGTAGTATTGCCTATCATATGCCAGTAGTTTTAAGTTTAGAAGGGGCTTTAGATATTGATG

[0141] TGTTAGAACAATCCTTTAGAGGAATTATATCAAGACACGAAGTGTTACGAACAATGATTAACTCTGAAGAAGGAGTAGGTTATCAAGAAGTGATTT

[0142] CAGCAGATAACTGGAGATTAGAAAAAGGTATAATAGAAGATAAAAAATATATTGATGAAACGATAGCTGCTTTTATAAAAATACCTTTCAATTTATC

[0143] AAAAGATTATAAGTTTAGGGCTTGTGTATATGATTTAGGAACAGATGGCTACTTATTAGCAGGAGTATTGCATCATATTGCCAGCGATGGTTGGTCA

[0144] GAAGGAGTATTCGTAAACGAGTTTAAAGAATTATATAGCGCTTTCTACAAGGGGAACACACCAATCTTGCCAGAATTACCCTTGCAATATTCGGATT

[0145] ATGCTATCTGGCAACGTAATCATATAAAAGGAGAGGTATTAGAAAATGAATTATCCTACTGGGATCAAAAATTACAAGGAGTCACTCCTTTGTCATT

[0146] ACCTACAGATTATGCACGTCCCTCTGTGCAAAGTACAGAAGGGGCTACAATAGATTTTCAACTAAATGAAGAGTTAGGAAATGCTTTAGAAACATT

[0147] GTGTCAACAAGAAGGTGTTACCTTGTTCATGATGTTATTATCAGCTTTTAAAGTTCTACTACATAAGTATAGTAATCAGGAGGATATTTGTGTAGGTA

[0148] GCCCAATAGCAAATCGTACGCAATCTGAGTTAGAAGGAATTATTGGTTTTTTTGTAAATACTCTGGCATTACGTACCGATGTAAGTGGTAATCCTAA

[0149] CTTTAAAGAGTTGTTACATAAAGTGAAAGAAACAACTTTAGAAGGATATGATCATCAGCTAGCTCCTTTTGAGAAAGTAGTAGATAGAGTGGTAGA

[0150] TACTCGTGATATGAGTATTACTCCATTGTTTCAGGTATTATTTGTACTACAGAATACTCCAGAGATTCATGGTGAAATGCTAGAAATGCAAGATGTGT

[0151] CTATAACCAATTATGAGTTCGGTAATACCACTTCCAAATTTGATTTAACCCTTAATGTACTAGAAAATCAAAAAGGTATTTTATTAACAATGGAATAC

[0152] TGCACTGCATTGTTTGATAGAACAACCGTGGAGCAAATGTTGATACATTACAAGGAGTTGCTTATTACTATTACAAAGAATATAGAAGCGCCTATAC

[0153] ATAAATTGTCTATTATAACAGAACAGGAGCGAGATACATTACTGTATACATTTAATAATGGTATCTGTGAATATCCTAAAAATAAAACCATAGTTGAT

[0154] TTGTTTACCAATCAGGCAGAAAAAACTCCTGATGCAATTGCTGTGGTTTATGAAGGACAGAAGCTGACTTATAAAGACTTAGACGAGCGATCTAA

[0155] TCAATTGAGCCATTATCTAATCCAAAAAGGAGTACAACCTGAAGAATTAATAGGTATTTGTATGGATCGATCTTTAGATATGATTACCGGTATTTTTG

[0156] GAATACTAAAAGCTGGCGGTGCTTATGTTCCTATTGATCCGGATTATCCTCAAAACAGAATCGATTATATGATAAAAGATTCTGGAGTTAATTTAGTA

[0157] GTAAGCAACTCTGATATAACACCTGTTTTTGAAGAAACAAAAGGATTGAGTATCGTATCTCTGGATAAAGAATGGGGAGTTATCAGTAACAATTCA

[0158] ACTCATCCAACGAATAGACAACTGAATCCGAATGGATTGGCCTATGTGATTTATACTTCTGGTAGTACTGGTAAACCTAAAGGAGTTTTAATAACCA

[0159] ATACCAATGTGGTACGATTATTCGAAAACGAATCTCCTTTGTACGATTTTAATTCTGATGATGTTTGGACATTATTTCATTCGTTTTGTTTTGATTTCT

[0160] CGGTATGGGAAATGTATGGAGCATTGTTTTACGGAGGACGTTTGGTTATTGTACCTAAAACTGTAACCAAAGACGCAATTGCTTTCAAGAAGTTAT

[0161] TGATAGAGGAAGGAGTTACTGTGTTAAACCAGACTCCCGGAGCCTTCTATGTACTTCAGGAAGAAATACTACAGGAAAATCAACAAACCTCATTG

[0162] AGATATGTAATTTTTGGAGGAGAAGCTTTGAATCCCAGTTATTTATCTGGATGGAAACATACCTATCCAAATTGTAAATTAATTAATATGTATGGAATT

[0163] ACGGAAACTACCGTACATGTTACTTATAAAGAGATTGAAGAAGTAGATACACTTAGTTCTACAAGTACTATTGGAGCTGCAATCCCAACATTAAGT

[0164] TGTCATATACTTGATGAACATTTAAACCTGGTTCCTATTGGAGTAGTGGGCGAACTTTGTATCGGTGGCGCAGGAGTAGCCAGAGGGTACCTTAAT

[0165] AGAGAAGAGTTAACATCAGAACGATTTATTCCTGATCCTTTTAGTACAGAGAAAGAAGCCCGGTTATACAAATCTGGTGATTTAGGAAGATGGTTA

[0166] CCCGACGGAACTATTGAATATATAGGTAGAAAAGATGCCCAGGTAAAAATTAGAGGATATCGAATGGAACTCGGGGAGATCGAGAATGTATTATCA

[0167] GTATTAGAATCTGTAAAACAATGCTGCGTTCTGGCCAAAGAAGATAGTGCCGGAAATAAACGTTTGGTAGCTTACGTAATTATAGAAGGGGATTTT

[0168] GATAAACTTGGTGTACAGACTGAGTTAGGTGAAAAATTACCAGATTATATGATCCCAAAGTTGTGGGTTGTTCTGGATCAAATGCCTTTGACAAGT

[0169] AATGGTAAGATTGATAAGAAATCATTACCAGAACCCGATGCAAATCAATTATCTACAAATGAATATGTAGCGCCTGAAACAACAACAGAAAAACA

[0170] ATTAGCAACGATATGGAAAGAATTGTTAGGACTGGAAAAAGTAGGAATACATGATAATTTCTTTGAGGTAGGGGGTGATTCGATTATTTCTATACGA

[0171] TTGATTTCTAAAATCAATGAAAGTTTTAGTAGGCAAACTCAATTACAGGATTTATTTAAACATAATACCATTAAAAGTTTTTCTGATGCAGTATTATT

[0172] GAGTGATCAACAAAACGATACATTTGAACTCTTGTATAAAACGGTTGAGGCAGAAATAACAGAATTGAGCCATACCGTTCAAAGTGAAATAGAAC

[0173] TTTCTGATCAGATAGAAGATGTATATCCTATGAGTGATGTACAGCAAGGGATGATCTTAGAATCTTTATTAGATCCCTCTCTTGCAGTTTTTCATGAT

[0174] CAGATGGCATTTCCGATTAAGGATACTTCTTTTAACATAGATGTATTTCAGAAGGCGATTACTTTACTGATAGAAAAACATGCCATATTCCGTACCGG

[0175] ATTCAATTTTTTGGATTACGATCAACCTATTCAAATCATTTATAAAACCGCTCCTGCTAAGGTATTGTTTGAAGATTTAAGTTTACTGGATCAACAAG

[0176] CACAAGAAAAAGCAATTGAGAACTATCTTGAATCTGAAAGGAACATCCCTTTCGAAATTAAAGCAGGACCTCTCTATCGTTTTAAGGTATTAACAA

[0177] TCGATGATTCTAATTTGTTATTCATTTATCAGTTTCATCATGCTATAATGGATGGTTGGAGTGTGGCTTCTTTCATTACAGAATTGTATAGAACTTATTT

[0178] CGAACTTAAAAAAGATATCCATTATACTCCTGCCGTCATAGGTGATGGTCAGCGTGAATATGTGATTAGAGAATTAGTAGCCAAAAAAGATGCCGA

[0179] TGCTATTTCTTTTTGGAAAGAGAAATTATCAGAATACAAGTATTTGGATGTTTTTTCAGATACTAAACAACCTACACAATATTACCATAAACACTATA

[0180] GTGTATCCTATTTGAAGGACCTTAGAACGCTATGTAAGAAGTATAAAGTGTCGCTGAAGTCTTTGTTTTTTGGAGCGTATGTATATGCTTTAAAGAT

[0181] GTTGACCCATGAAGAAGAATTAACCGTTGGTTTGGTAGGTAATAATAGACCGCAAATCAAGGATGGAGATAAGACATTAGGATGTTTTTTAAATAC

[0182] ATTGCCTGTTAGGTATACTTCTCAATCAGAGCATAGTTGGCTGTCTTATTTTGAAAGTATTACGACTATACTTGAGGAAATAAATCTGTACGGTCATT

[0183] TGACTTTTTTTGAGATGAAACGCCATCTTGGATTACAGAAAGAAGATCTTGTTTTTGATACCTTGTTTAATTTTATCGATTTTCATGTATATAATGAGA

[0184] TTTTTGAAAAAGATGCTTCGCCAGATAGTTTGATAAGAGATGAAGGGGATAAAAAAGGATTAAAAGTTAGATCTTTTGAGAGAACCAACTCAAGT

[0185] TTGAATTTGACTGTAAATTTAACCGGTGGTAGAGGAGTAGAGTTTTCGTATGAACTTCATAATGATTTTAAGTCCGGAATTACCTTAGAACATTTTC

[0186] ACACGTTCATTGATAATATTCTGACCTGTTTTTGTAATAAGCCGGAAGAACCAATCGATAATAAAACTTTATTATCGTATTCAGAACAGCAGAAATT

[0187] ACTTCAGGATTTTAATGATACTCAAATAGCTTATCCAAAAGATAAATCAGTTGTTGATTTATTCAGAAAACAAGTGGCAAAAACACCAAACGGTAT

[0188] TGCTGTTGTTTCTGAAAAAAGTACTTTAACATATAAGGAACTGGATGATCGTTCTAATCAATTGGCAAATTATATTCTTTCAAATTATACTATTGAAA

[0189] AAGAGAATTTAATAGGGGTAATGTTGGACCGAAGTGATCAATTAATTATTTCAATACTAGCTATCCTGAAAACCGGATGTGCCTATGTTCCTATTGAT

[0190] ATTAATTATCCAGAGGAACGAAAAGAATATATAAAGAAGGATAGTGAATGTGTCTTAATTGTTGATGAGGTAGTATTGAATTCTTTCAAAGAAGATC

[0191] AAGATAAATACTCAGAAAATTTTTCAACTGTAACTATTGAACCAGGTTATTTAGCCTATGTTATTTATACTTCAGGATCTACAGGGAGACCTAAAGG

[0192] AGTTATGATAGAACATCAAAATTTAGTGCATTTATGTTTCTGGCATAAAGATGCATATAATGTTACTCAGGAAAGTAAAAGCACATTGTTTTCGAGTA

[0193] TTGGTTTTGATGCTTCTGTCTGGGAAATTTACCCATACTTACTTTTTGGAGCTAGTTTATACCCTATTTCAGAAGAGCTTAGGTATGATTTGGATCAG

[0194] CTACCAAATTTTTTGAAAGATCACAGGATCACTCATAGTTATATTCCTACTTCATTATGTCAAAGTTTTATAGATCAGGAAATTTCCTTACCGTATACA

[0195] ACTGTTTTGACAGGTGGAGATACGTTGCATATAAGTAAGCCGACAGATATTGTTCTATATAACAATTATGGCCCAACTGAAGGTACTGTGGTTGCCA

[0196] CCTATCATAAAGTAACTAATGGTCATGATGATCAAAAAATACCGATAGGAGTTCCTATTTCTAATACGGAGGTTTACATTTTAAATTCGGAGCTTGAT

[0197] TTATTACCAATAGGAGTTATTGGAGAATTGTGTATTAGCGGTTCAGGTTTGGCCCGGGGGTATTTAAATCGTGAAGATTTAACAGAAGAGAAGTTT

[0198] GTTGCTCATCCATTTAAACCGGGAGAGCGTTTATATAAGACAGGAGATTTGGCTCGCTGGTTAGCTGATGGCACTATAGAGTTTATAGGCCGTAAG

[0199] GATACCCAGGTAAAGATTCGTGGTTATCGTATAGAATTAGGCGAGATAGAAAGTGTTTTATCAGATCTATCAATTGTTCAGAGTTGTTGTGTATTGG

[0200] CCCGGGATGATGGTAATGGCAGTAAACGTTTAGTCGGTTATGTTGTTGTAGAAGGTACATTAGAAAAGGATGAGTTAGAGCAGGAGCTACACTTG

[0201] AGGTTACCGGATTATATGGTTCCCCAGTTATGGATTGAGTTAGAAGAGATGCCATTGACCAGCAATGGTAAGATAGACAGGAATGGGTTACCTTCA

[0202] TTAGATGTTTCGCAGTTGTCGTTACAGGAGTATGTTGCTCCCCGTACGGAGATTGAAACACAACTGTCGGTTATCTGGCAGGATTTGTTAGGAGTT

[0203] GATAAGATTGGAGTATATGATAATTTCTTTGAGTTAGGAGGTCATTCGTTATTAGCGACACGTTTGGTTGCTATGATCCGCAAAGAGCTGTCTGTCG

[0204] AAGTTGCTATTCGAGATATATTTTCACATTCTACGATTTCGGACTTAGGACATCATTTATCTGATGCTTCAGAAAACACAGTTTTACCTTCGATTATA

[0205] CCACAGGATAAGCCAGAACATATTCCATTATCCTTTAGCCAGGAGCGATTGTGGTTTATAGATAAGTTAGAAGGTAGTATTGCCTATCATATTCCTGC

[0206] AGTTTTACATTTAGAAGGGGATTTAGATATAGCTATTTTAGAAGCTTCCCTAAGAAGTATTGTTTCTCGTCATGAAGTATTACGAACGGTATTTTATG

[0207] CAGAGGATGGAGTAGGATACCAAAAAGTGATTGGATCTGATGATTGGGTATTGCATAGAGAGCTAGTAACCGATACGACAAGTATAGATGATAAGA

[0208] TCGCTGCTTTTGTAGAGGATTCTTTTGATTTGTCAAGGGATTATATGTTTAGGGCTTGCTTATATGATTTTGGATCGAATCATTATATACTGGCTGGGG

[0209] TTTTTCATCATATTGCCAGTGATGGTTGGTCAGAAAGTATTTTGATTAATGAGTTGGTAGAATTGTATAGTTCAATTCAGGAAAAACGTTCTTCGAA

[0210] ATTACCGAGTCTGAGTTTGCAATATACCGATTATTCGATTTGGCAACGAAAGTATATTGAAGGTGATATTTTAGAAGGTCAATTATCATATTGGGAGG

[0211] ATAGGCTGAAAGGAGCTACTCCATTATCCTTACCAACAGATTATGCACGACCTTCGATACAGAGTTATGAAGGGAATAGTGTTTCTCTTACGTTGGA

[0212] TGCTGATTTGAATAGGATGCTATCAGAACTTTGTTCAGAAGAGGGGGCTACTTTGTATATGGTTTTATTATCAGCTTTCAAGGTATTATTATCCCGTTA

[0213] TAGTGGTCAGGAAGATATTTGTGTAGGTACTCCAGTAGCCAATCGCACCCAATCGGAGTTAGAAGATATGATAGGGTTCTTTATCAATACGTTGGCA

[0214] CTTCGAAGTGATCTGGAAGGAAACCCGAGTTTTAAAGAAGTTTTAGAAAGAGTAAAAGAAACTACGCTAGATGCATATGATCATCAATTGGCTCC

[0215] TTTTGAAAAAGTGGTAGATAAGGTCATAGCGACTCGTGATATGAGTATGAGTCCTTTGTTTCAGGTAATGTTTGTATTACAAAACACACCAAAAGT

[0216] AAAAGATCTTGAATTTAGTAAAACACGTGTTAGCACCTACGAGTATGAAGAAACCACTTCTCAGTTTGACCTGACCCTGACTGCGGTAGAAGAAT

[0217] CTGGAGGTATTTCATTAAACATGACATATTGTACGGCTCTGTTTAATAAAGATACAGTTACTCGTATGTTAGTTCACTATAAAGAATTATTGAGTAGC

[0218] CTTGTCAAAGATATTACGTTGCCTATTCAAGCGTTGTCTATGGTAACAAATCAGGAGAGAGAAGAATTGCTATACAAATTTAATGATACAGTAGTAG

[0219] CATATCCAGAAGATAAGAGCATTATTGATTTGTTTAAGCAGCAGGTCAAAAAAACTCCAGATGCTATTGCTGTCATTTTTGAAAAAGATAAACTTA

[0220] GTTTTAAAGAACTGGACGAAAGATCTAATCAATTGGCGCATTATCTTATTAGCCAAGGTATCAGATCAGAAGAATTGGTAGGGATCTGCTTAGATC

[0221] GTTCTTTAGAAATGATCATAGGTATTTTAGGGATACTAAAATCAGGAGCAGCATATGTACCTATAGACCCAGAATATCCATCAGAGCGTATCGATTAT

[0222] ATATTGGAAGATGCTACCATTAAAATCTTATTGAGTAGTATCGACAGAAAAACCCTCTTCGAGAAAACACATACATTAGAGGTTATTTCATTAGATG

[0223] AAGATTGGGGTGCTATAAGCAATGAATCTACTGGCGATGTAGATAAGCAATTTAACCCAAAATGTTTAGCCTATGTAATTTATACTTCGGGTAGTAC

[0224] AGGAAAACCCAAAGGTGTGATGAATGAACATAGAGGTGTTGTTAATAGGCTATTATGGACTCAATCGCATTATAATCTTGATCCAAAAGATGCTATT

[0225] CTGCAAAAAACATCATTCTGTTTTGATGTATCGGTATGGGAATTGTTCTGGCCCATTATAAGTGGAGCGAAATTAGTTTTTGCAAAACCAGAAGGG

[0226] CATAAGGATGCACGGTATTTAAGAACATTGATAGAATCACATAACATTACTACAATTCACTTTGTGCCATCTATGTTACGAGCCTTTTTAAGTGAGAT

[0227] AAATATGGGAGATTGTAGTTGCCTAAAACGTGTTTTATGTAGTGGAGAAGCTTTGCTGGTAGATCAGGCACTATTATTTAGAAAGAAATTCAGTAAT

[0228] GTTAGATTAGATAATTTATACGGGCCGACAGAAGCAGCGATTGATGTGAGTTCCTGGGAGGTTCCGGCAGAAGGAGATTTGTCAAGAATTCCTATT

[0229] GGGAAACCCGTTGCCAATACAAACCTTTATATTCTTGATAAAGAGGATCAGGTATTACCAATAGGAGTGGTAGGAGAGTTGTGTATTGGAGGAATT

[0230] CAGGTCGCAAGAGGATATTTAAACCGGGAAGAATTAACAAAAGAAAAATTTGTAGCAGATCCTTTTATCGTAGAAGGCCGTATGTATAAGACAGG

[0231] AGATTTAGCCCGATGGTTAGCAGATGGTACTATAGAATATATAGGAAGGAAAGATGATCAGGTCAAAATTAGAGGATACCGTATAGAATTGGGTGA

[0232] GATAGAAAATGTGCTATCTACAATTGAAAATATTCATAGTTGTTGTGTTCTGGCCAGAGAAGATGGCAGTGGGAATAAACGCCTGGTAGGTTATGT

[0233] AGTAGGAAATCTTGATAAAGAAAAAGTCCAAAAAGAATTGTATTCAAGATTGCCAGAATATATGGTTCCTCAGTTATGGGTCGAATTAGAAGAAAT

[0234] GCCGCTGACCAGTAATGGTAAGTTAAATAAGAAAATACTTCCTGATCCGGATACATCTCTATTATCATCCCAGGAGTATGTTGCACCGCGTAATAATA

[0235] TAGAGAAACAACTGGTTGAGATCTGGCAAGAATTGCTTGGTATTGATAAAGTTGGAGTGTATGATAATTTCTTTGAGCTGGGAGGGCATTCGTTAT

[0236] TGATAGTTCAGTTAATTTCGAGACTACATGATCAAAACCTCCAAATAGAGGTTAAAGATATTTTTGCAAACCCCACAATAGAGGCATTAGCATTAA

[0237] GGTTAGGTACAGTTACTTCTCACTATGAAGTTCCGGCCAATGGTATTACAGTAGGCTGTAAGAAGATTACTCCAGATATGGTTCCATTACTTGATTTT

[0238] AGTCAGAAGGAGTTGGATATGGTTGTAGATCATATTGAAGGAGGCGTTGCTAATATCCAGGATATGTATCCGCTTTCTCCATTACAGGAAGGAATGT

[0239] ATTTTCATCATTTATTGAGTGACCGTGATAGTGGAGATCCTTATGTTTTGCTTACAGGGATTTCTTTTGATCATCCCGATAAACGTTCAGAATTTATAG

[0240] AAGCGTTCCGCTATGTAGTTACTCGTCACGATGTACTTCGTACCTGTATTGTAAATGAAGGATTACCAAATGCCGTTCAGGTGGTTTTACGTGATGT

[0241] GAAACTTAAGGTGTCAGAATTATCTTTTTCTTCATCGCAAAATGTGAAAGAGGAATTAGAATTGATGATGGCTTCGGGGAAACACTGGATAGATTT

[0242] GTCAAAAGCTCCTTTATTAAGTTTAGAGACAGCAGATGATCCTACCACAGATAGTTATTACCTAATGATGAAGTATCATCATACCGTGATAGATCATG

[0243] TTGGTTTAGAGAAGATCGTAGGAGAGATTCAGGTTTGTTTGTCTGGGGATAAATCAAGCCTTTCGACTCCTGTTTTATATCGTGATTTCATTGGCCA

[0244] TGTATTATATCAACAATCAACTAATGATAGTGAGGTGTATTTCCGTTCACGTTTTGAAGGTATTACAGAACCCACGTTTCCTTTTGGCTTGTCTGATA

[0245] CATTAGGAGATGGTAGTGGTATTGAAGAATCTCGTTATGTTTTACCTCGGGATTTATCAAATCAACTTCGATCATTATCCCGTAGTTTAGGTATGACT

[0246] CCCGCGGTTCTTTTTCATGCAGCCTGGGGATTAGTGATAGGTAGATGCAGTAGTAAGGATTATGCAATTTTTGGATCATTATTTTCGGGTCGATTACA

[0247] AGGCTCATTAGAGGCAGGGCAATCTTTAGGACTGTTTATTAATACACTTCCCGTGATCTTGAATTTATCTGATAATGTACGAGATTATGTAGCAGAA

[0248] GTAAAGTCGGAGTTAGAAAGTCTATTACCTTATGAGCAGACTCCTTTATCACGAATTCAGAACTGGAGTGGTGTTTCTAATGATACTCCATTGTTCA

[0249] GCGCCTTATTAAATTACCGTCATTCTTCTCCTGCTTCTGATAATCCAGACACAGATTTAGGGATGCGTGTATTAGGAGAGAAAGAACGAACTAATTA

[0250] TCCTTTTGAATTAAGCGTAGATGATTTGGGAGAAGATTTTAGTTTGACTGCTTTTGTTGATGGTGATATTACACCTTTACGTGTTATTAAGTATATGG

[0251] AAATGGCCTTGCAGGAACTTATAAAAGGCCTTGCTAGTAAAGATGTTATTGAGGTAAATAGCTTGAGTATCTTACCAAAAGAAGAACTACAATTAT

[0252] TAAATGGTTTTAATACTACAGAAGTATCCTATCCATTAGATCAAACGGTAGTTGATTTATTTTCTGCTCAGGTTTCCCGTTCTCCATCAGCAGTAGCT

[0253] TTATCCTATGAAGGTTTAGAATTAACTTACCAGGAGTTGGATGACCGATCAAACCAATTGGCTCATTATTTACAAGAAAAAGGAGTTGAGAATGAT

[0254] GAGCTGGTAGGTATTTGTATCACTCGTTCTTTTGAGATGGTTATAGGTATTTTGGGGATTTTAAAATCCGGTGGGGCTTATGTTCCTATTAAACCTGA

[0255] TTTTCCACTTTCGCGTATTTCCCATATTGTAGAGGATACAAATTGCGATTTGATTTTAACCGATAGCTTGAGTTTATCATCTTTGGACCTGTTGGATGT

[0256] TACCAAAATTATATTAGATGAAGATACCCCAGAGTATTCAGGATATCCTACTGCTTCTCCAGATATTACCATCGTTCCTGATGCTTTAAGTTATGTGAT

[0257] TTACACATCCGGTAGTACAGGAGTTCCCAAAGGAGCTATGATCGAGCATCGTGGGTTGTTAAACCACCTGTTGCTTATGATAGATGAGTTAGAGAT

[0258] GGATTCTACCAGTGTTGTTGCTTTTACAGCACCTTTTACCTTCGATATTTCAGTATGGCAACTATTGAGTGCTTTATTATGTGGCGGTCGTGTTGCTA

[0259] TTTATAATGAAGATATAATTTTAGACCCACAGTTTTTAGAGAAGTCTTTATCAGAAGATGAAGTGTCGATACTTCAATTAGTTCCTTCGTATGTTTCA

[0260] GAATTATTAGATGTCGAAACGCCTAAAGGCCTGGATAAATTGAACTATTTTTTAGTTACTGGTGAGGCTGTTTCGCGAGGAGTATTATCAAAATGGT

[0261] TTTCTCAGTACCCTGCCATTCCCGTTGTTAATGCCTATGGACCTGCCGAGGCAGCAGATGATGTTACTTTGCATAAGATGTATGAAGTACCAGATGG

[0262] TTCGATGATTCCTATCGGTAAGCCCGTAGCCAATATGCAGATTCATATTCTGGATGCTGTTCTAAATCGGTGCCCTATAGGAGTGATAGGAGAGATTT

[0263] GTGTTAGTGGAGTAGGAGTTGGACGAGGATACATCAATGATGCAATGAAGACTGCGAGTAGTTTTATTTCAGATCCATTTGTAGAAAGAGGTCGTA

[0264] TGTACCGCACAGGAGATCTGGGCCGTTGGTTAGCAGATGGTACCATTGAGTTCGTTGGTCGTGCAGATGATCAGGTAAAAATCAGAGGGTATCGT

[0265] ATAGAGTTAGGAGAAATAGAAAATGTATTATCAGGTATAGAAGGTATTACTCAGTGTTGCGTTTTAGTTCCCGATGATCATAACGGGAATAAACGTT

[0266] TGGTTGGTTATGTTGTTTGTTTAGGAGCTTTTGATAAAGTTTTGATTCAGGATTTTTTATCATCACAGTTACCATCGTATATGGTTCCCCAATTATGGG

[0267] TTGAATTAGAAGAGATGCCATTGACAGGTAATGGTAAAATAGATAAAAAGTCGTTACCATTGTTAGATTCTTCAGTTTTGTCTGGCGAATCTTATGT

[0268] GGCCCCAAGAACAGATATAGAAGAACAATTGGCATTGATCTGGCAGGACTTACTTGGTGTTGATAAAGTTGGGGTGTACGATAACTTCTTTGAGCT

[0269] AGGAGGGCACTCATTATTGGCTACCCGATTGGTGTCTAGACTTCGAAAGGAGATGAGTATTGAAGTTTCCATCAGAGATGTATTTGTACATAGTAG

[0270] ATTAGAAGATTTGGGAACTCATTTATTATCCCACCGTTCTGGAGTTATATTACCAGCCGTAGAGGTTCAACAACGTCCGGATCGCATTCCTTTATCGT

[0271] TTAGCCAGGAACGTTTATGGTTTTTAGACCAGTTAGAAGGTAGTGCTGCTTATCATATTCCGATAGTTTTACGGGTAGAAGGTAGTTTGGATATTGAT

[0272] ATATTAGAGACTTGCTTTAGAACTATTGTATCTCGTCATGAAGTGTTGCGAACGATGATCCTTTCTGAAGATGGTATAGGTTATCAAGAGGTGATTG

[0273] AGTCAAATAATTGGAAATTAGATAAGGTTAAAGATCAATCAGACACTTCTCTGGAAACTTCTTTAATGTCTTATTTGAATATTCCTTTTGATTTAGCG

[0274] AGAGATTATAAGTTTAGAGCTTGTCTATATGAATTAGATCCTGATCGTTATGTTTTGGCTGGAGTTTTTCATCATATTGCCAGCGATGCGTGGTCAGA

[0275] GGATCTATTAGTAAAGGAATTTACAGAGTTATACGAGGCATATCAGGAGAATCGTGAGGTAGCCTTGCCAGAATTATCGTTACAATATGCAGATTAT

[0276] GCGATTTGGCAACGTAACTATGTAGAAGGCGAAATATTAGAGCGTCAGTTATCATATTGGGAGACTAAGTTACAGGGAGTTGCTCCTTTAGCGTTG

[0277] CCAACAGATTATGTTCGCCCATCAGTTCAGGATTATAAAGGTTCATGTATTACCTTGAATTTGAATCCGGATTTAAGTTCAGCAATTAAAGTAGTATG

[0278] CCAACAAGAAGGAGTTACCTTATTTATGATGTTGTTATCTGCTTTTAAAGTTTTATTGTACAAGTATAGTGGCCAGGCTGATATCTGTGTGGGAACTT

[0279] CGATAGCCAACAGAACCCAGTCGGAGTTAGAAGACATGATTGGATTTTTTGTTAATACGTTGGCACTGCGAAGTAATTTAGGAGATAATCCTGATT

[0280] TCAAAGAATTTTTATCACAAGTAAAAGATACGACTTTACATGCTTATGATCATCAATTAACTCCATTCGAGAAAGTAGTAGATCGAGTTGTTGATAA

[0281] AAGAGATATGAGTATCAATCCATTGTTTCAGGTAATGTTTGATATGCAAAATACTGCTAATGAAACTTTAACGTTAGAAGGGATAACACTATCTCCAT

[0282] ATGAGCATGAAGCAAGTACATCGCAATTCGATTTGAATCTGGTTGCTGAGGAGAAGGAATCAGGTATTTTATTACGACTAGAATATTTTACTTCGTT

[0283] GTTTAAAGAGTCTACAGTAAGGCGCCTATTATCTCATTATGAAGAACTGTTAAAGAGTATTGTCAATAATATTTCTTCTCCTATAGATGCTTTATCGAT

[0284] GTTGCCAAAGCAAGAAGAAGAATTAATCTTAGGTCGAAGAGTTACACCATCAGGACTAAGTTTTAATCCAGGAGCGGTTGATCTGGGTAATGAAT

[0285] CTCCGATCAATGTTCGTTTTGAATCTATAGTAGCCACTAATCCTGATGCTGTAGCCTTAGTACATAATGAAGAGGTTTGGAATTATAGCGACCTTAAT

[0286] TCGTATGCAAACCAGATTGCTCATAGTTTGTTGTCGTTGGATCTTTCGTCTTCTTCTATAGTTGGAGTGTATATGGATCGTAGTGTCGAATTTGTAGG

[0287] TTGTTTACTTGGTATTTTCAAATCGGGTCATGTGTATACTCCTTTGGATACGAACAATCCTCCATCCCGTATAGGGTCGATGTTATCGGATAATCCATT

[0288] TTCTGTACTCATTACAACAAGTTCTTTATTATCAGAATTAGGAGATTTGAAAGATATCATAGTTTTGGTGATTGATGAATTGTCTGATGATTCTATAGA

[0289] AGATAAGGTCGTTTATGATAGCAATATGATTGTAGGTAAACCTACGGATAACCCTAAGAATAAGAATTCAATGGATAGCTGGGCGTATATTTTATATA

[0290] CATCCGGATCTACTGGAGCTCCCAAAGGTGCCATTACACGTCACGATGGAGCGATGAATCATATTTTAGCAGAGTATGCTGTTATGGATTTACCGGA

[0291] TGGTTTCCGATTTTTACAGAGTGCAGGTATTGGTTCAGATATTTCAGTATGGCAGATTTTAGGTCCTTTACTCAAGGGAGGAGTAAGCGTAATTGTC

[0292] GACAAGTATGAGTTATTAGAGTATGGCAAGTTGTTGGATACCTTGATTCGAAGAAGAGTTACTCTTATCGAGTTTGTACCAACCTATATGTGGGGAT

[0293] TATTGTCGTATATAAAAGAACAGGAAGAAGCGGTTATTTTGAAAGACCTATCAAGCATTATGCTTGTGGGAGAGTCTATTCCTGTTGCGATGGTTAA

[0294] TGAACTGAAACGTTTGTATCCCGAGGTAAGATTATGGAATGCTTATGGCCCTTGCGAGGCATCGGATGATGTTATCCAGTATGAGATCAGCGACTTT

[0295] TTAGAGGAGACCCAGGTACGAGTTCCTATCGGTCGTGTTATTCCTAATATGAACACTGTTATTTTGGATAAATCGGGAGGGTTATGTCCTATAGGAA

[0296] TTATAGGAGAGATAGGTGTTAGTGGAGTAGGAGTAGGAGCAGGATATTTAGGATTACCAGATCGTACTTCAGAGAGTTTTATAGAGAATCCGTTTG

[0297] GTGATTTATTAGGTGATGTTTTGTATAAGACAGGAGACTTAGGTCGTTGGCTTGAAGATGGGAACATTGAATTTATAGGAAGGGAGGACCACCAG

[0298] GTTAAGATTCGAGGACATCGTGTAGAGTTAGAGGATATAGCTTCAGCCTTACGTAAGGATGCTTATGTTGAGGATTGTCATGTTTTGGTTCACAGG

[0299] ACAGGAGGTCAGGATTTAGTGATTTGCTTTGTTGTATTGAGTTCTCAGGGATTATTATATGCTTCAGATTCTTCAATTACAGAAGTTTTCTATTTGTTA

[0300] TGCAAAGAAGAGTTACCGAGTTATATGCATCCTTCTCATTATCATATTGTAGATGAGTTTCCATCGAATTTGAGTGATAAAGTAGATGGCAAGGCTT

[0301] TGTTAGATATTTTCTTATCAGATTTTGATGATGGAATTAGTCTTTCTCAACATGTTTATGAAGCTCCACGTAATGAAATAGAGGAACACCTGGTTGA

[0302] GATCTGGCAGGAATTGTTGCATATAGACAAAATCGGAGTTTATGATAATTTCTTTGAGTTAGGCGGTCATTCATTATTAGCGACACGATTGGTGTCTA

[0303] TGATCCGTAAGCAACTAAATATAGAAGTAGCAATAAAAGACATATTCGAATTTAATACCATAGATGAATTAGCATCACATATAGAATTTATTAACCTG

[0304] GATCTGGATGAAGATCAAAATGAGGATACAGAACAATATAACGTAACCATAGAAATATAATTCTACTTAGTCACCTTACCAGCAACTTATCGGAAAT

[0305] GCGTTAGGGATAGTAGTGTAAAGCCCGGAATCTGATGATAATCGGATGAGGACTTGTAACGTATAGCCCGACCCGAAGGGAAACGCCCAAAGATG

[0306] ACGAAGTAACAATGACTTTAGCTATAACTGAGCTATACTATTTAGTCAAGATCAATCAAAATAAAGATTTAACTACTCCATGAAATATAAAACCGGA

[0307] GTTAAACGATTAATGGCAAATTCTATCTGCTGGCCGGTAGATGGATCTGTCCTTTAAAGCAATATTATAAACACATGAAAAGAGAAGTTTTAAATAT

[0308] TTTAAAAGAAGCCAATAAAAACAACATAAAATTGGTAGTTAATAAAGGGGCGTTAACTGTTAAATCTACAGATCCAATAGATGCAGATTTACTTCA

[0309] AAAAATTAAAGATAATAAGGAGTCAATTATTCAATATATAGAGAGGTATCAAGGAGAAAAAAAGGGGAATGCTTTAAAAATTAAACCTTACAATAG

[0310] AGATGCTATCAACAAAATTCCTTTGTCCTTTAGTCAGGAACGTTTATGGTTTTTGGATCAGTTACAAGATAGCCAGGTTTACCATATACCTGTAGTTT

[0311] TAAAACTACAAGGAGTATTGGACTTGGATATATTAGAAAGCTCTTTAAAAACGATTATTGCTCGCCATGAGATATTGCGAACTGTTATTTATTCTGA

[0312] AGAAGGAATAGGTTACCAAAATGTAATTTTGGAAGATCATTGGACTTTAGAAAAAAATATGATAGATGGGGGAAAAAATATTGAAGAGGAAATAG

[0313] CTTCTTTTATCGAACTCCCTTTTGATTTATCAAAGGATTACAAGTTTAGAGCTGTTGTGTATGATTTGGGTGAAAATCTTTATGTGTTCGTAGGGGTT

[0314] TTTCATCATATCGCCAGTGATGGTTGGTCAGAAGGAGTATTCATCAATGAGTTTATAGAGTTGTATAGTGCACTACAAGAGAATAGAACCCCTGTAT

[0315] TGCCAGAATTATCATTACAATATGCAGATTATGCTGTATGGCAACGATCATCTATTGTAAATAAAACAATAGAGGAAGAACTTTCTTATTGGGAAGA

[0316] GAAATTGAAAGATGTTACTCCTTTATCATTACCTACCGATTATCTACGACCATCAATTCAGAGTTATGAAGGAGCAAGCATAACTACTACATTAAAT

[0317] GTCGACTTATGTCGTTCTTTACAGGATTTTTGTCAGAAAGAACAAGTTACTTTATTCATGACTTTACTGTCAGGCTTAAAAGTAGTATTGTCTCGATA

[0318] TAGTGGTCAGGATGATATTTGTATAGGTACTCCTATTGCCAATCGTACCCAATCAGAGCTAGAAGATATGATTGGATTTTTTGTAAATACTTTGGCAT

[0319] TACGAAGCGATCTGACAGGCAACCCAAGTTTTGAAGAAGTTGTTTCCAGAGTTAAAGAAACAACATTGAATGCTTATGATCATCAATTGGTTTCTT

[0320] TCGAACGTGTCGTTGATCGTGTTGTATCCACACGTGATATGAGTATGAGCCCATTGTTTCAGGTATTGTTTGTGTTACAAAACACTCCTGAACAAA

[0321] CAGATTTATCCTTAGAAGAGATTGCAATTTCTACATATGATTATCAAGAAACTACTGCTCAATACGATATTACATTTAATGTTATAGAACATGATTCTG

[0322] ACATTTCATTACAGGTAGAGTATTGTACAGCGTTATTTGATAGTGCTACAATAGAACGTTTGTTATGTCATTATGAGGAGTTATTACAAAAAGTAGTT

[0323] AAAGACAGTAAAATACCGGTTAACTCCATATCATTGTTAGGGGTAGAAGAACGAGAAACATTACTTCATAGTTTTAATGATACCAAAGTAGCATAC

[0324] CCATCAGAACAAACATTAATAACTCTGTTTGAATCTCAGGTAGAGAAACATCCAGAAAGGGAAGCGGTGTTTTTTGAAGGGAAATCGTTGACGTA

[0325] TAGGGAGCTGGATGAACGTTCCAACCAATTAGCACGTTACCTTCTTAATCAAAAGATCGAACCAGAAGAATTAATAGGAATTTGCTTAGAGCGTTC

[0326] GATAGAAATGATCGTTGGTATTTTAGGAATCTTAAAAGCAGGAGCAGCGTATGTACCGATTGATCCAAATTATCCATTGAGTCGTATCGAGTATATG

[0327] CTTGATGATGCAGCTATTGATATGGTAATGACAACAAGTGATATTGTATCTGTTTTTGGTGAAAGAGAAAAATTAAACACAATTTTATTAGATGGGC

[0328] ATTGGAAGATTATAGAGCAGTTGGATGTGAGTGCAATACCCGAGCAGTCTAATCCTGAACATCTGGCATATGTTATTTATACGTCAGGAAGTACGG

[0329] GCCTTCCTAAAGGAGTTATGATTGAACATCGTAGTATTGTTAATACCATTTGTTCACAAGTAGAAGCTTTTTCAATTACAAAGGAGGATAGATGCCT

[0330] TCAATTTGCAAGTTCTTCTTTTGATGCTTCTGTCTGGGAAACTTTTATCTCATTGCTTGGAGGAGCTTCGTTATATATTGCGACCGAGGAAAGTAAG

[0331] TCAGATGTAACTTCTTTTGGACAATTTATCAAAGAACATTCGATTACGGTTGCTACTTTGCCCCCTGCATTTTTACAGATACTACCTGTTGATGCTTT

[0332] ATCGAATGTACGTACCTTAATTACAGCAGGAGAGGCTATATCTGTAGATTTAGCTAAGGCATTTTCTGAAGGACGTACCTATATTAATGCATATGGTC

[0333] CTACAGAGAGTAGTATATGTGCCACCACATTTTCAGGAATTATAGAAAATCGTGTGCCTATCGGAGTTCCTATTTCTAATACTGAGGTTTACATTTTA

[0334] AATTCGGAGCTTGATTTATTACCTATAGGAGTTGTTGGTGAGTTATGTATTAGTGGTTCAGGTTTGGCTCGCGGGTATTTAAATCGTGAAGATTTAAC

[0335] AGAAGAGAAATTTATTGCTCATCCATTTAAATCGGGAGAGCGTTTATATAAGACAGGGGATTTGGCCCGCTGGTTATCTGATGGTACTATAGAGTTT

[0336] ATAGGCCGTAAGGATACCCAGGTGAAGATTCGTGGTTATCGTATTGAGCTTGGAGAGATAGAAAGTGTTTTATCAGATCTATCAATTGTTCAGAGTT

[0337] GTTGTGTATTAGCTCAGGATGACGCTAATGGCAGTAAACGTTTAGTAGGTTATGTTGTTGTAGAAGGCACATTAGAAAAGGATGAGTTAGAGTGGG

[0338] AGCTACACTTGAGGTTACCGGATTATATGGTTCCCCAGCTATGGGTTGAGTTAGATGAGATGCCTTTGACCAGCAATGGTAAGATAGACAGGAATG

[0339] GGTTACCTTCATTAGATGTTTCGCAGTTGTCGTTACAGGAGTATGTTGCTCCCCGTACGGAGATTGAAACACAACTGTCGGTTATCTGGCAGGATT

[0340] TGTTAGGAGTTGATAAGATTGGAGTATATGATAATTTCTTTGAGTTAGGCGGTCATTCGTTATTAGCGACACGTTTGGTTGCTATGATCCGCAAAGA

[0341] ATTATCTGTCGAAGTTGCTATTCGGGATATATTTTCACATTCTACGATTTCGGGCTTAGGGCATCATTTATCTGATGCTTCAGAAAACACAGTTTTAC

[0342] CTTCGATTATACCACAGGATAAGCCAGAACATATTCCATTATCCTTTAGCCAGGAACGATTGTGGTTTATAGATAAGTTAGAAGGTAGTATTGCCTAT

[0343] CATATTCCTGCAGTTTTACGATTAGAAGGGGAGTTAGATATAGCTATTTTAGAAGCTTCCCTAAGAAGTATTGTTTTTCGTCATGAAGTATTACGTAC

[0344] GGTATTTTATGAAGAGGATGGAGTAGGATACCAAAAAGTGATTGAATCTGATGATTGGGTATTGCATAGAGAGCTAGTAGCCGATATGACGAGTAT

[0345] AGATGATAAGATCGCTGCTTTTGTCGAGGATTCTTTTGATTTGTCAAAGGATTATATGTTTAGGGCTTGTGTATATGATTTGGGAGGTTCGACTTATG

[0346] TATTGGCAGGAGTTTTTCATCATATTGCCAGTGACGGTTGGTCTCAGGGTATTTTGATACGAGAATTTCTTGAGACTTATAAAGCTATTAAAGCAGG

[0347] AGGTTCGAGAACCTTACCAGAATTACCTTTACAATATACCGATTATTCGATTTGGCAACGAAAGTATATCGAAGGGGATATTTTAGAAGATCAATTA

[0348] TCGTATTGGGAGGATAAACTAGGAGGAGTTACTCCATTATCCTTACCAACAGATTATGCACGACCTTCGATACAGAGTTATGAAGGGAGTAGTGTT

[0349] TCGCTTACGTTGGATGCTGATTTGAGTAGCATGCTATCAGAACTTTGTACAGAAGAGGGAACTACCTTGTATATGGTTTTGTTATCAGCATTCAAGG

[0350] TATTATTATCCCGTTATAGTGGTCAGGAAGATATTTGTGTAGGTACTCCTATCGCTAATCGTACGCAATCTGAGTTAGAAGATATGATTGGTTTCTTTG

[0351] TTAATACACTCGCACTTCGTAGTGATTTAGGAGGAAATCTGAGTTTTAAAGAAGTTTTACATAGTGTAAAAGAGACAGCATTAGCCGCTTATGATCA

[0352] TCAATTAGCTCCTTTTGAAGAAGTGGTTGATAGAGTCATAGATACCCGAGATATGAGTATGAGTCCTTTGTTTCAGGTGATGTTTACATTACAAAAC

[0353] ACACCAGAAGTAAAAGATTCTGACTTGGATGATATAAGAACTTCTCCTTATAATCTTGAAGAAACCACTTCTCAGTTTGACCTGACCTTGACCGCA

[0354] ATAGAAGAGGCAGGAGGTATTTCTTTAAGTATAACGTATTGTACAGCTTTATTCACTCAGGGTAGTATGGAGCGTATGTTGGCTCATTATAAAGAGC

[0355] TGTTGAGTAGCCTCGTTAAGGATGTTACTCTACCCATCCATTCGTTGTCCATGATGACAAATCAGGAGAGAGAAGAATTACTGTATGGGTTTAATGA

[0356] TACGATAGTAGCATATCCAGAAGATAAGAGCATCATTGATTTGTTTAAGCAGCAGGTTGAAAAAACTCCCGATGCTATTGCTATTGTTGATGAAGAT

[0357] GTTAAGATAACATACAAAGAACTAGACGAAAGATCTAATCAATTAGCGCATTACTTACTATCAGAAGGAGTGCAGGAAAAATCCCGATTAGGGCTT

[0358] TTATATGAGAGAGGAATGGATATGGTGATAAGCATGATAGGTGTGTTAAAATCAGGTGCTACTTATGTTCCTCTCGACACTTCTCTACCACCGAATC

[0359] GTTTGGATTTTATTTTGAAAGATGCTGAGATAAACTACATTATTCATAGTGATTCTTCTTTATTAACAGAATTATCCTCTGATGAATATACTTTTTTGAA

[0360] TGTCCAAAAATCTTTAGAACATTCTATTGATGTATATAACTCAAATAGAAGTATAGTATCTGATACCTATATAATGTATACCTCCGGAACAACCGGTAC

[0361] TCCAAAAGGTATAGTGATTAGTGACGAAAATGTAATAACGCTTATCAATGATAAAGAGAGTTCAATCGCTATCCATCCATCAGATCGGGTATTGCAA

[0362] TGGTCAAATTATGCTTTTGATGGTTCTACATACGAGATTTTTGGAAGCCTTTTAAATGGTGCATCATTGTATTTGATCCCAGGTTCTATAGCTTCTGAT

[0363] GTTGATGCTCTAACTCAGGTTATTAGTAAAAATCAATTAAGCGTAGTTTTTATAACTACAGCGTTGTTTAATTCTTTTGCAGATTATGACTTGTCAAG

[0364] ATTGTCGAGTTTAAGAATATTATTATTCGGAGGCGAAAAAGTTTCTGTTAGTCCAACAAGAAAAATAGCATCAGCTTTAGGGCAAGAAAGATTACT

[0365] TCACGTATATGGTCCGACCGAAACTACTGTATATGCAACTTGTTACCCAATTTGTGATATTCCTGATAATGCAGGAACTATTCCTATTGGAGGCCCGC

[0366] TTACTAATACGAGCCTTTACGTTTTAGATACCAATATAGAATTAGCCCCTATGGGTGTAGTTGGTGAGTTATGTATTGGAGGGGCAGGTTTAGCCAA

[0367] AGGGTATTTGAACCGACACGAATTAACCGCAGAGAAATTTGTAGTTCACCCTTTTAAACCAGGAGAACGTTTATATAGAACGGGAGATTTAGCCC

[0368] GATGGTTACCTGGCGGAAATATAGAATTTATTGGGCGTAAGGATAATCAGGTTAAAATTAGAGGATATCGTATAGAATTAGAAGAGATAGAAAGTGT

[0369] ACTGTCTTCTGTAGAAGGTATTCAGAGTTGTTGTGTATTAGCACAAGAAGATGCTAATGGCAGTAAACGTTTGGTAGGATATGTGGTTGTAGAAGG

[0370] ACAGTTGATTAAGGAAGAATTACAACAAAAATTACACTCTGGATTGCCAGAGTATATGGTTCCTCAATTATGGGTCGAATTAGAAGAAATACCTTT

[0371] AACCAGCAATGGTAAGATTAATAAGAAAAAACTACCTGTTCCTGATCTATCTTTGTTATCCTCAAAAGAATATATTGCCCCTCGTAATGAGATGGAA

[0372] GAGCAATTAGCTTTGATCTGGAAGGATTTACTAGGGATTGACAAGATTGGAGTGTATGATAATTTCTTTGAACTGGGAGGACATTCATTACTCGTAG

[0373] TAAAACTCATCTCCCGATTAAAAGATCAGGAGCTTCAAGTAGGAGTTCGTGATATCTTTGCCAATCCAACAATTGGAACATTAGCATTGCGATTAG

[0374] GAGTTGCTGCCCCAAGTTATGAGATTCCTGCCAATGGTATCACAACAGATTGCGAGAAGATTACTCCCGATATGGTCCCGTTACTTGATTTTAGACA

[0375] GGAGGAGTTGGATATGGTTGTAGATCATATTGAAGAAGGCGTTGCTAATATCCAAGATATGTATCCGCTTTCTCCACTGCAAGAGGGAATGTATTTT

[0376] CATCATTTGATGAGTGATCAGAGTAATGGAGATCCTTACATATTATTTAATTTGTTGTCTTTTGATAGCCTGGATAAACGTTCACAATTTATAGAAGC

[0377] ACTCCGCTATGTGATTACACGTCATGATGTACTTCGAACTTGTATTTTGAGCGAAGGGCTACCTCATGCTGTTCAGGTAGTTCTAAGAGAGGTTAA

[0378] GCTTCAAGTATCAAAATTATCTTTTTCTTCATCGCAAAATGTAAAAGAAGAACTAGAATCAATAATTGATTCTGGGAAACATTGGATGGATTTATCA

[0379] AAAGGGCCATTGTTACAGTTGCAAACTGCTGATGATACAAGTAGTGGTCGTTTCTTCTTACTATTTAAACAACATCATGTAATAATAGATCATGTTG

[0380] GTTTGGAGAAGATAATAGAAGAGATTGAAATGTATTTGTCTGGAGACACGGTTAATTTTACCCAACCGGTTTTATATCGCGATTTTATAGCACATGT

[0381] TTTACATCAACAAGCGACTAATGATAGCGAATCTTATTTCCGTTCACGTTTAGGAGATATCACAGAACCTACTTTTCCTTTTGGTTTGTCTGATACAT

[0382] TGGGAGATGGTAGCGATATACAGGAAGCAAGTAAGATGCTTTCTTCAGGTATATCTGCTCAAATTCGTTTACTATCTCGCAAATTAAAGATAAGCCC

[0383] AGCCGTAATTTTTCATGCCGCTTGGGGTATTGTGATAGGTAGATGTAGTAATAAAGAGTATGCCGTTTTTGGAACTTTGTTTTCAGGACGACTACAG

[0384] GGATCATTGGGAGCAGATCAGTCTTTAGGGTTGTTTATTAATACGCTCCCCGTGATACTACATTTAGATCATAGTGTATCAGAATATTTAGATCATGT

[0385] AAAACAAGAGTTACAAGGTTTAATGCCTTACGAGCAAACACCATTATCTAATATTCAACATTGGAGTGGTGTTTCAAATGATACTCCGTTGTTCAG

[0386] CGCATTGTTAAATTATCGTCATTCAGAACCCCAATCTATATCAGATAATGTGGGTATTGATTTAGGAATAACACTTCAGGTAGACAAAGAACGTACA

[0387] AATTATCCATTTGATTTGAGTATTGACGATTACGGAGTTGATTTTGGTTTAACAGCCTGTATAGATGGGATTGAGCCGGATCGGGTTATTGCATACAT

[0388] GGAAACAGCTTTAATAAAGCTTCTTGAGGAGATTGAATGCCCAAATAACGTTGAGGTGAGTAGTTTGAGTATTTTGGCAGCCGAAGAACTACAGG

[0389] TATTAAATAGTTTTAATGTTACAGATATTGGCAATCTTCAGGATAAAACTATTCTGGATATATTCGAAACACAAGCACAACAGACTCCTGATGCAGT

[0390] GGCAGTTGTTTACCAAAACGAAAAGATTACCTATAGAGAATTAAATGAAAAATCAAATCAATTAGCACATTATATAAGTAATCAAAACATTTCTGA

[0391] GAGTGCTTTAATCGGTGTTTGCGTGTCTCGCTCTATAGAAATGATTGTCGGTGTTTTAGGAGTTTTAAAATCAGGAAATGCCTATGTTCCAATTGAC

[0392] CCGGAATATCCTTTGACTCGAATTTCTTACATCATTGAAGACGCTAAGCTTGATCTTATAATAAGCACTACTTCAGACATAAAACAATTAGATGTTAA

[0393] TGAGGAAGTGAAGTGTATATTGTTTGACAGAGATCACGAATTAATTACCAAGGAGATCACTACAACACCCGGGATAAAATTGACTCCCGAACATAT

[0394] GGCATATGTGATTTATACGTCAGGAACAACAGGTAAACCAAAAGGAGTTGCTATAACTCATCTCGGTTTGTACAATGTATCATTGAGTTGGCAAAA

[0395] AGAGTACGATTTGGATGAGAATACTCGTTTATTACAGATAGCCAGCTTTTCTTTTGATGTATTTACAGGAGATCTTTGTCGTAGTCTATTATTTGGAG

[0396] GGCAGATGATTATTTGCCCTGGCGCTATTCGCTTTGATCTTCGAGAACTGTATAAATTAATGGCTAGCCATAAGATCAATATAATGGAGGTAACTCCT

[0397] TCATTAGGAATACCATTAATGGATCATATCTATGATAATGAATTAGATTTTGAGTGGCTTAAACTCTTTATTTTAGGATCTGATGTTTGTACTTCTGTC

[0398] AATTTTTGCCGACTAGAAAGTCGATTTGGAAAACAAGTAAGAATTATAAATAGTTATGGAGTTACAGAAGCTACCATTGATAGTTCTTATTATGAGG

[0399] CCGAAGATCTAAATAGCCTGGATAGCTTGACTAATGTTCCTATTGGTAAACCTATGGGGAATACTCATTTTTACATTTTAGATGCTTTTAGAAATGTA

[0400] TTGCCTATAGGTGTTATAGGAGAACTATATATCGCGGGACCAGGTCTGGCAAAAGGATATTTACATAAAGAAGAACAGACAAAAGAGAAATTTGTT

[0401] TCAAATCCATTTGTAGAAGATACTTTAATGTATCGTACAGGAGATTTAGCACGTTGGTTACCTGATGGAAATGTAGAATTTATTGGTAGAGGAGATA

[0402] CACAGGTAAAAATTCGAGGATATCGAATTGAGTTAGGAGAGATTGAAAATGTATTGGCATCTTATGAAGCCGTTATAAATTGTTGTGTTTTAGCTAA

[0403] AACCGATACGCACGGAAATAATCGTCTTGTAAGCTATGTTGTTTTAAAAGAGAGTCTGGATAAACAGGCTGTAGAAGAGTATTTAAAAGAAAGCA

[0404] TACCAGAATATATGATTCCACGTATTTGGGTGGAGTTAGAGAAAATGCCTTTAACACCAAATGGCAAGATAGATAAGAAATCGTTACCCGATCCCG

[0405] ATAATTTGGAGTTATCAACCCAGGAATATACTGCTCCTGGTAATGAACTGGAAGAACAATTAGTTGATATTTGGCAAAAACTTTTAGGAATAGAAC

[0406] GAATAGGAGTGTATGATAATTTCTTTGAATTGGGGGGGCATTCTTTATTGGTAACAAGATTAGTTTCCATAATTCGTAAGGAATTGAATATCGAAGTT

[0407] TCTATTCGAGATATTTTTGAGTATACCAACATACATGAATTGGCTCTTCATTTAAAACAACAATCAGTAGGTGTTTTATTACCCGAAATTGTGGTTCA

[0408] ACAACGTCCGGAACGTATTCCTTTATCATTTAGCCAGGAACGTTTATGGTTCTTAGATCAGTTAGAAGGTAGCGTTGCATATCATATTCCTATAGTAC

[0409] TACGATTATCAGGTAGTTTGGATATTGATATATTAGAAACCTGTTTTAGAACTATTGTATCTCGTCATGAGGTATTACGAACAATGATTCATTCTGAGG

[0410] AAGGTATAGGTTACCAGGAGATTGTTGATTCTGATGCTTGGAAACTGGATCGGAATATGGATCAATCAGACGTTTCGTTAGAAGATTCTATAGCATC

[0411] TTATTTAACCATCCCATTTGATTTGTCGAAGGACTATAAATTCAGGGCTTGTATTTATGATCTGGGATCAGATCGTTATGTTTTGGCCGGAGTTTTTCA

[0412] TCATATTGCCAGCGATGCATGGTCAGAGGATATATTGGTAGCAGAGTTTGCAGAGCTCTATAGTGCATATCAGGAGAACCGTACGGTAGCTTTACC

[0413] AGAGTTATTATTACAATATTCGGATTATGCGATTTGGCAACGCAATTATATAGAAGGTGAAATATTAGAGAATCAGTTGTCTTATTGGGAAGAGAAG

[0414] TTACAGGGGGTTACACCTTTAGGACTACCAACAGATTATGCTCGTCCATCAGTGCAGAGTACAGAAGGAGCGACCTTTAACTTTGAGTTAGACAA

[0415] AAAGTTAAGTGATTCAGTTACCGAATTATGCCAAGAAGAAGGGGTTACCTTATTTATGGTGATGCTATCCGCTTTTAAAGTTTTATTATATAAATATA

[0416] GTGGCCAGTCTGATATTTGTGTGGGAACTCCCATAGCTAATAGAACTCAATCAGAGCTAGAAAATATGATTGGGTTTTTTGTTAATACATTGGCATT

[0417] GCGAAGCAATTTAAAAGATACTGCTGGTTTTAATGAATTGCTACAAGAAGTAAAGAGTACAACGCTAGAGGGATATGACCATCAATTGGCTCCTTT

[0418] TGAAAAAGTAGTAGATCGGGTAGTCAAAACTAGGGATATGAGCATGAATCCACTGTTTCAGGTGCTGTTTACACTGCAAAATATATCTGCAGAAAC

[0419] AGATTTAGTAATGCATGGTGTAACTATGTCGCCTTTTGAATATGAAGGAAACACTTCACAATTTGATTTAACCATTAACATAACAGAAGAAGTTACT

[0420] GGTATTTCATTTAATGTAGAATATTGTACAGCTTTATTCAAGAAAGAGACTGTAGAGCGTATGTTGATGCACTATCAAGAACTATTACATAACATTGT

[0421] AGCCAATCCATCAGAGAAAATCGGAAGCCTTTCTATGTTGAGAAAAGAAGAAAAATCTCAATTGGTAGATGCATTCAACGCTACAGATGTTTCTTA

[0422] TGATCAAAATAAAACCATCGTAAGTCTATTCGAAGAACAAGCCAATAAAACTCCGGATGCTGTTGCTTTAATCTTTAAAGATAAGCAGCTAACATA

[0423] TAAAGAGCTTAATGAACAATCCAACCGCTTGGCATATCATTTAAGGGATACGTATAGTCTGAAAGCAGATGATATCGTCGGTATTTTATTAGATCGAT

[0424] CTGATTGGGCTGTGATTTCAATTTTAGGGATACTAAAAGCTGGTGCAGCCTATGTTCCTATTGACACCAGTTACCCAACATCTCGAAAATCATATATT

[0425] GTGGAGGATAGCACTCCTAAGGTTTTGATTACAGACTCCGAACATCTTTTTGATAGTATAGAGTTTAATGTAAGTATATTTTCGATAGATGTAGAATT

[0426] AGAAGAACTACTTGGTGACGCAGAATTATTGACCAATCCTTCTTTTATATCTTCTCCAAAAGATCTGGCATATATTATTTATACTTCAGGAAGTACAG

[0427] GTAAGCCTAAAGGAGTAATGATAGAACATCGCAATATAACTAACTACGTGTTATATGGTTTATCGGCATATCAATTAGAAACAGGAAGTCATAATTT

[0428] CCCATTATTTACTTCCCTGGCTTTTGATTTAACACAAACCAGTATCTTTCTCAGTTTACTAACTGGTGGTACCCTGTTTGTTTATGAAAAAGAAGCA

[0429] ATAGAATCTTTACCCTTAATTTTGAAGACCAAAGAAATTACCTCTATTAAATTAACGCCATCTCATTTATCATTTTTAGAGGATACAGAGCATGCTCA

[0430] TTTGAAACGTATTATTATAGGTGGAGAAGCCTTAAAAGGATCGCATATTGATTTGTTAGGACCACTGGATAATTCGGTTAAGGTGTTTAACGAATAT

[0431] GGCCCAACCGAAGCTACAGTTGGTTGTTCTGTTATAGAAGTTAGCACATATGATACTTCTTTATCATCTGTACCCATTGGAAAACCTATAGACAACA

[0432] CGCAAATCTATATACTAGATGACCAACAACAACTGTTACCTGTTGGAGTAATGGGAGAATTGTGTGTTGGAGGGAATGGACTGGCAAGAGGATAT

[0433] CTGAATAAGGAAGAACTAACACAAGAGAAATTTATATCCAATCCATTTGTAGAAGGAGAACGTTTGTATAGAACCGGAGACTTAGCAAAGTGGTT

[0434] ACCCGATGGAAATATTGAGTTTATTGGTCGAAAAGATGATCAGATTAAGATTAGAGGATATCGTATCGAATTAGGAGAGATAGAAAATGTTTTGAG

[0435] CCAGGAAATAGATATTCTCGAATGTTGTGTGTTATCAAAACCAGATGCTGAAGGAAATCACCGTTTGATCGGATATGTAATTAGCGAAAATGATTTT

[0436] GATAAAGATAGATTACAATCCTCATTGCTACAGAAACTACCTGGTTATATGGTACCAGGTCTTTGGGTTGTTCTTGAAACGATGCCTCTAACAAGTA

[0437] ATGGTAAGATAGATAAAAAAGCATTACCTGATCCTGAACTGATCGATATTGCCGCAGCTAATTATATCGAACCTCAAGAAGAAATGGAAATCCAGT

[0438] TAGCTGCAATCTGGAAAGAGCTATTAGAGGTAGAACGAGTAGGGATTCAGGATGACTTTTTCTCTTTAGGAGGACATTCCTTGCTGGCTACTCGTT

[0439] TGGTATCAATGATTCGAAAAAAATTGAATATTGAAATAGCAATTATCGATGTTTTTGAATATCCGATATTATCAGATTTATCAAAGTATGTAGATTCTC

[0440] AATCCCGAGGTAGAAGTATACTGCCAGAAATTGTTGCTAGAGAAAATAATGCAAAGATCCCATTATCATTTAGTCAGGAACGTCTGTGGTTTTTAG

[0441] ATCAATTGCAAGGAAGTGTAGAATATCATATTCCGTTTACCCTTCGTTTTACAGGAGATCTGAATACTTCAATATTAGAAAATGCTATTCAAGAAGTA

[0442] ATTTCTCGTCATGAAGTTTTACGCTCTGTTATTGAAGCTGAAGAAGGAATAGGGTATCAAAAAATAAGGCCTGCTACCGATTGGTCTATGGATTTG

[0443] GTTCAGGTAGAAAATGAGGAGATGATTCTTGAAAAACTAGAATCATTTATCGATACTCCATTCGATATGGCAGTCGATTATATGTTTAGAGCCTGCT

[0444] TATATCAATTAGATGAAAATGATTATGTATTCTGCGGAGTTCTTCATCATATAGCAAGCGATGGATGGTCTCAGGGTATTTTGATAAATGAGTTTACTC

[0445] ATATATATAGTGCATTACAAAAAGGTCAATCATTTATTTTACCAGAGTTACCAATACAATATGCAGACTATGCGATATGGCAACGAGAATACATAGAA

[0446] GGTGAAATCTTAGAGCAACAGTTGTCGTTTTGGGAAACCAGGCTTCAGGGAGTTCCTCCTTTATTATTGCCTACCGATTATGATCGTCCTTCAGTTC

[0447] AAAGTGTTGAAGGAGCATCTGTTAACTTCGAATTAGATGCAAAAACAACTGAAACCATAAAAGAATTTTGTAATGAAGAAGGAGTAACCCTGTTT

[0448] ATGTTCTTATTATCGGTCTTCAAGGTATTGCTTTATAAATATAGTGGTCAGGATGATATCTGTATAGGAACACCTATTGCAAACCGAACACAAGCAGA

[0449] ATTAGAAGGGTTGATAGGTTTCTTTATCAATACATTAGTGCTCAGAAGTGATCTAAGTGATAATCCAGAGTTTAGAGAGCTATTACAGCAAATAAAA

[0450] TCAACAACACTCGAAGGGTATGACAATCAATTAGTGCCTTTCGAAAGAGTTGTAGATCGCGTTGTAAAAACCAGGGATATGAGTATACCTCCTCTA

[0451] TTTCAGGTAAAATTTATGTTAGAGAATACTCCCGAATCAGAAGATTTACATCTCGAAAATTTGGAAATCAACTCATATGAGTATGATGCTGAGTTTT

[0452] CTCCATTGGATATATCTTTAAATGCAGTGGAGTCAGGAGACGGTCTATACTTTAATTTTGAGTATAGTATTGCGTTGTTTGCCAAAGAAACGGTACA

[0453] AAGAATGGCATTGCATTACAAGGAGTTAATACAGCATATACTGGTATCTCCGTCATCTAATATAGGAAGCTTAACTATGCTATCAAATGAAGAGTCT

[0454] TCAATACTGTTAGATGTATTTAGTAGGTCTAATACCACTTCAGTTTCTTATGGAGAGGATAGTACATTAGTCTCGTTATTCGAAAAACAAGCCGCCG

[0455] AAACACCAGAGGCAATTGCAGTAGTTTTTGAAGATAGAAAGTTGACATATAAAACCTTAAATGAAAAATCAAATCGTTTGGCACGCCACCTAAGA

[0456] GAGAACTACGAGTTAAATTCTGGTGATTTGGTAGGAATACAATTAAACCGTTCTGACTGGTCTGTTATTTCAATGCTGGGTGCTATGAAAGCGGGA

[0457] GCTGCCTACGTTCCGATTGATGTAGAATATCCTGCTTCGCGAAAAGCGTTCATTATTAAAGATACACAGCTAAAAGTGTTAATCATAACTTCAGAAA

[0458] AATTATTTGATAGCATAGAACTTAATGTGCCGGTTTTTTCAATTGATATAGAGCTAGATGATTTATCTCAAGAAAAGGCATATCTACAAAACCCTGAT

[0459] TATTCTGTAACACCTGGTGATTTAGCGTATTTGATTTATACTTCGGGGAGCACCGGGCAACCAAAAGGAGTGATGATTGAACATCGGGGTATTGTC

[0460] AATACCATATGTTCTCAGGTCGATGCATTTTCGATTACTGCAAATGACCGCTGCCTTCAGTTTGCAAATCCGTTTTTTGATGCTTCTGTATGGGAAAT

[0461] TTACATTACTCTTCTGGGAGGTGCTAGTTTACATATACCATCAGAAGAAGAAAAAGCAGATACAGTTTCTTTTGCAAAATTTATGAAAAAGCATGA

[0462] GGTAACATTTGCCACATTGCCACCTGCTTTTTTACAATTATTAGAAAGCGATAGCCTTCCGACCATAAAAACCTTGGTAACAGCAGGAGAAACTAT

[0463] ACCATTAGAAGTTGCCAGAGCCTTTTCTGATTCCTGCAAATATATAAACGCTTATGGTCCAACCGAAACCAGTATTTGTGCAACAACTTTTGAAGG

[0464] AGATATTACAGGAGGTAGCGTTCCTATAGGAAAGCCGATCACGAATGCTAATGTTTACATTTTAGACGCAGATATGAATTTGATGCCTGTTGGTGTT

[0465] CCTGGTGAATTATGTGTAGGAGGACCAGGATTGGCCAGAGGATATCTTAATCGTGAAGAATTAACAAATGAAAAGTTTGTGGATAATCCTTTTGCA

[0466] GAAGGGGAACGATTGTATAAAACCGGGGATTTAGCTCGATGGTTAGCAGATGGTAATATTGAGTTTATGGGAAGAAAAGATGATCAGGTAAAAATT

[0467] AGAGGATATCGTATAGAATTAAGTGAGATCGAAAGAGTACTTCTAGAAAAAGAAGAGATCCAGCATTGTTGCGTAGTAGTAAGAGGTAGTACAAT

[0468] CGATACCAGGAGTTTAATAGGCTATGTCGTTGCACAAGAAAACTATGATACTAGAGATATCAATGATTATTTGAAAGAAAGACTTCCAGAATATATG

[0469] ATACCGGGGGTATGGGTTTCTATGGATGTATTGCCGATGAACAGAAATGGTAAAATCGATAAAAAAGCTTTACCAAGTCCAGAAGTTTCTGATATAT

[0470] CTAAAGCAGAATATGTGGCACCACGAAACGATACAGAAGTGAAATTAGTTGCTCTATGGGAAGAGTTATTAAGTATAGAAAGAATTAGTGTACATG

[0471] ATGATTTCTTTGCTTTAGGAGGCCATTCTATCCTGGCAATACAATTAATATCTCGCATTAATTTGTTATTAAATACATCGCTTAGCGTTTCTGTTATATT

[0472] CGAGAATCCTACCATTTCAAAATTCGTTCAGAAGATGTCTTCTACAGACTTTTTTACGTATAAGGTCATGGTAACCTTACAAGGAAATGGAAGTAA

[0473] AAAACCAATATTTTTTGCCCCTCCGGCTGCAGGTACTGTAGGTTCTTATTTACAATTATCCAAAGCACTAGGAGATGATCAACCAGTGTATGCGTTT

[0474] CAATGCCCGGGATTAGACGGAAAATCAAAAACATCTGAATCTGTAGAAACCATGGCAGAAACGTTTATTACAGCAATGCAAGAAGTAGATCCTTC

[0475] TGGCCCATATCGATTAGGTGGGTATTCTTTTGGAGGAGCCGTAGCACTCGAAATGGCATTACAATTAAGAAATAAAGGATTTGAGGTTGAAGAATT

[0476] ATTGATGATTGATGCTAAAATTCATGAAGTAAGTGATGAGATATATGAACAAATAGATGAAGCACAAAGATTCCGCGAATTACTTTACGAAGAAGT

[0477] AGATGAGTTAAGAGAGATCTTCGATTCTGAGTTAACCAGATCAAAATTAGCACTCGAAGGTAAATCTATACCCGAGCAGATCAATGTAATTTGTGA

[0478] ACTGGCCAGTGGCTCAGAATTTTCTGTAAGCGATGATGATATTAAAGGGCAATTAGAAGTTCTTTACAGTAATTCGACCTATAGATATATACCGAAT

[0479] CTCGATCAAAAACTGGATACTCAAATCACATTATTTAGAGCAATGTATGTTCCAAAAATATTGGAAAATTCAGAAATGGAAATGATAAATAATGATG

[0480] CAACAGAATTTGATTATGGATTACAGCAATATACCAGTAAGAAGGTCGTAATACATGCTATACCAGCTGTACATACTACCATACTAGAAGAAACGCA

[0481] TGCAAAAGAAATTTCAGAGTTTCTTAAAGTCGATAACATGATAGAAGTAGCTGATCAAATGATATAATTTTTAACCTTCCATATCCAGCAAAACTTA

[0482] CCAATAGTAGTAGATAACTAATGAAGAAATACTTTTTATACGTAGGCCTGATAATGATTTCATTATTAGGTACCGATTTAACCGATTTTGCCTTAAGTA

[0483] TTTGGGTGTTAAAACAACCAGAAGGAACAATTACTTCATATACTATGATATGGTTTTTTGAAGCGGCTCCTGCTGTTCTTTTAGCACCATTTATTGG

[0484] CAGTTTTGTAGATAGATGGAGTAAAAAGAAAATAATCATTTATGGGCAGATCATAACAGGTATAGGTAGTAGTATACTGATGTCACTTTTTTATTTCG

[0485] AACATTTACAACCCTGGCATATTATGATCGTATCAGGAATAGGTTCTATAGCCAGTATGTTTGTTTTTAGTGCTTTTTATGTCGCTACAACGGCATTA

[0486] GTATCAAAAGATAAACTGATAAAGGCGCAGGGAGTATCAACTGCAATTTATGCTGCTATAAGTATTGGTGTTCCTGTGGTTTCTCCGGTATTATTCA

[0487] AAATGATCGGTATGGAACGCATATTTATGATTGATATTATCACTTTTTTGATCTCGATCCTTGCTTTTTTGATGATACGATTTGTTGTTGTTGCGAAAA

[0488] CAGAAGAAGAATTCAATTTTAGAAATGATTGGAAATTAGTGAAAGAATTTCTAAAAGAGCGAAAAGGATTAAGATCACTATTAGGTTTTTTCTTTA

[0489] TAATCGGTTTTTTAATTGGACTGATCAAAGTACTATTTACACCTCTGATATTGGATTTTTCAAATGAGTACTATTTGGGCTTGGTATTATCTGTAGTAG

[0490] GAATTGGTACTGTGATAGGAGGAGGAATAATGAGCATGAGTAAAAAGATCGAACATCCTATTAAAACTATCTTTAATGTTAATATCCTGTTAGGTGT

[0491] CATTCTTGGTTGCTTATGGATTAATGTTAACCTTTATGTTTTGGCTATTGGAGGAATGTTGGCTGTTCTTTTGTTTACCATTTCCGATATTATGAATAAT

[0492] GCTTTTTTTCAAGCTGTCGTTCCAACAAAAATGTTAGGAAGATTATCAGGTTTTGTTGGATTAGTCATTGGGGCTTCAGAGCCATTGTCTTATATGC

[0493] TATCGGGGGCTTTTGTAGATTATCTAAGTAATGTTGTTTCTGATGTTTCTTACGAATTTTCAAATCACTTCCCAGGAACCACTATTACAATTGCCATC

[0494] ATCCTCATCTTTACAATATCTGGAGTAATGCTAATGATTGTTTCCTATTTGTTTCAAAGAAATAAGGGGATAAAAAACCTGGATGCTTTATACCAGGA

[0495] AAAACTTAAGAAAGAATCAAAAGATCAATCAGAACATAATAAAAATACAGCACATATCAATTCTCCTAGTATGAAATGATGCTTTATCATAAACTTT

[0496] TAAAATCAGTAATAATTCAATGAACGAATTTCAAGTAATACAGGAAAAACAAATTCTTGAACATATACCTATTCGAAATGGTATAACCAGGAAAGA

[0497] ATTTTACAAGGATTTTGTAAACAAAGGACAACCAGTAGTTATTAGAGGAATGACTACAGGTTGGAATGCTTTAGAGTGGGATGTAGATTATTTCAA

[0498] AGCTAAAGAAACCGGAGTTAAAATTCCTATTAAAGTAGGTAAAGTGAGCGAAGGTAGACGAGAAATGATGCTGATTAGTCAATATGTCGAACTGT

[0499] TGGATACATACGAAAAAAACGTAAAACTTGGGTTAGATGCTGACCCGGTGGGATACTTACATGATATTCCCTTTTTTTATATGTTTCCAGAGTATTTG

[0500] GCAGATATATTACCTTTTCCAAAAGAATTGTTGCCCGAGTGGTATTGGCCAAAATGGCAAAACTATATTCAATTCTTCATGGGAGCGACAGGAAGC

[0501] CTTACCCCATTACATTTTGATACGCTTTTAACTCATAATTTGTTTTTTCAGGTTGTAGGTAGAAAGCAATTCATCTTGATTGATGGGGCCCAAAAAG

[0502] ACCAGTGTTACATAAAAGGATGGAGATGGTCCAAATTTGATCCCAAAAATCCGGATTATGAAAAGTTTCCTAAAGCCAAAGATCTAAATATAAAAG

[0503] AAGCGATTTTAGAACCTGGGGATATTTTGTATATGCCTTCAGGAATGTTGCATCAAGTCCATGGGTTAAGTCAATCAATTTCTTTCAATATAGATTGG

[0504] CATACCTCTAGAAGTGCTAAAAAAGGAGTGGGAACTTTATGGAAAGGAGCTCCTCTTAAAAATGTATACTATAATGCATTGGTGTATGCAGGATTA

[0505] CAACTCAAAGTTCCCGAAAAAAGCATATTTCCATTATATAAGTCATACCTAAATTATATCAGTTAATATTTAATATAAAAGAAATGAGTACAAAAGTA

[0506] TATCTTAAACCTAATGTTGTTATTGAACCCTTATTTGATAATTGGTATGCCTGGAGTCATTTGATTTCTCCTGCTACAGGAGCAATGAATGTAGTAGG

[0507] GCGACATTTAAAAATAATGAGCTCTTATATACAGGCCCCTGCTGCACATGCAGCAGCAGTTAAAAACCCAAAGATGCTGGGAGGACCTTTTATTGA

[0508] TTATGGAGGAAAGCGTGTTGATGAAATAAAAAACCTTAAAGAAGAAACTTTACAAAAACAAAAAGATTTGATTGCATTTAATGATGCCGTTAAAG

[0509] AATTAGATGCATTGTTAAAAAGTAAAGCCAAAGGTTTTTCGTTAGAGCCACTATATGATGAGGTGCCAGAAATATTACAAGGATATGTCGAGTTAAT

[0510] TTATGATTTGAATAACCAACCTTCTTTTAGGTTTTTTGAATCTCTGTTATATAAGAGTAAATTTTATAATCCGGATTCACAAAGTATTTTACTGTGGAT

[0511] CACAGAGAATGATGAACGCCCTTTCGTATTAAGTACACCTCGATTACCCGATAGTAATCAAGTGCATTTGGATATTCCTTTTGATCATAAAGGCTTG

[0512] GATAAACTAAGCAGAATGAAACGAATACCAGGTTCTGTAAGTGAAGTAAAAGAACTATTGGGGATATCAAAAGAACAGGAAGAAGTATTTGATAC

[0513] TTTTTTTACAACAGAACCTCCTAAAGAATACAATCAATATACCGGTGATAAGGTGCGTATGCGTTATTTTGGTCATGCAAGTATACTTGTTGAGACA

[0514] AAAGATATTTCTATTATGTTGGATCCTGTAATAAGCTATTATGGGTATCATCAAGATGTTTCGCGCTTTTCTGATATGGATCTACCCGAAGTAATTGAT

[0515] TATGTTTTGATTACTCATAATCATCAGGATCATATATTATTTGAAACCTTGATCCCGTTGCGTCATAAAATCAAAAATATTGTAGTTCCCAGATCTTCA

[0516] AATGGTAGTTTACAAGATCCAAGCTTAAAACTAATGTTACAAGCCATTGGTTTTGAAAATGTCATTGAAATTGACGAAATGGAAGCTATTAACTTT

[0517] AGTGACTGTACTATCACAGGAGTACCTTTTATTGGGGAACACTCAGATTTAAATATACAATGTAAAATATGTCATCATGTGGCTGTAGGAGATTTTA

[0518] AAATGCTTTTTGTTGCAGACTCTTGTAATGTAGCAGCAAAGTTATATGAACATGTACAGGCAGAGCTAGGAGATGTTGATGTGTTATTTTTAGGAAT

[0519] GGAGTGTGACGGAGCGCCTTTGTCCTGGCTATATGGGCCATTGTTAACAGAAACGATAGATAGGGATAAGGATTTTTCTAGAAGATTGGCCGGATC

[0520] AAATTTTGAAAGAGGAAAAGCACTTGTTGAAGCATTTAGCCCAAAAGAAGTATATGTATATGCTATGGGGCAGGAACCATGGCTGGAATTCATAA

[0521] GTAGTATCAAATACACAGATGAGTCTAACCCTATTATTCAATCTGATAAATTAGTTAAGTATTGTATGGATGGAAATATAATATCCGAGCGATTATTTG

[0522] GCGAAAAGGAAATTTTGTACAACAAGACACAGGAATCTAAAAAGGTTTTAGAATCTGTATAAAAGAAGGTACAATAACTGTAAAAAAAAAAAAA

[0523] GCCTTTGCTATAATTTTTCTGTCATCGTTTTTGATGAAATGATTAGAGTGTAGAACATACTTTATTTGGTTATTCTATGTACAGAAAAACATCGCTTTT

[0524] TATAGGCTAATCGGTAAATAAATCGATTAGCCTTTTTAAAAAATTGAAAAAAATGAATCATTTCTTTATTGCCAAACCTAAAATAATAGCTATTCCAT

[0525] TTGCCGGGGGTAATCAATATAGTTTTAAAACTATTGAAAAATACATCCAGAAAGATATCGAATGGGTTACCGTAGAGCTTCCGGGAAGAGGTGGAA

[0526] GATTCAGCGAACAGTCGTTAGATGCAATACATCTAATGGTGGCAGATCTTTTTGATCAAATATTACCTCATATAAAAGAAGGGCCATATATGATTTAT

[0527] GGACATAGTATGGGAACGCTATTAGGCTATGAACTAACCAAGAAAATAGTTGAGGCTAAAATGAGATTACCCAAATGTCTTTTTTTTACAGGACGT

[0528] TCTGCTCCATCAGTTTTGACTAAAAAGAAAAGATCAATACTACCCAAAACCTTATTTTGGAAAGAAGTATATGATATAGGAGGGTTACCAACAGAA

[0529] ATTTTAGATAATAAAGATCTTTTAGATCTCTATTATCCAATACTAAAATCTGATTTCAAGGCTATAGAAGATTATGAGTATATAACACAAGAGCAGCC

[0530] TTTTTCTTTTCCGATATATGTTTGTATGGGTACAGAAGAAGTAGGAGAAGATCATAATCAACAAACAATAGAAGAAATAATGGCTTGGGATAATGA

[0531] GACTAAGTCTTTATGTGGCCCCGAATTACTTGAAGGTGATCATTTTTTTATACTGAAGTCCCCTCAAATAATTGCCCAGAAAATTGTAAAAGCTTAC

[0532] AGAACCAATTCGGTCTTAAGGACTTCTTTTAAATAAGCTTCGACAGTATTAATACAATATAAACACATGAAAGTTTTAAAGTTTGGTGGAACCTCAT

[0533] TAGGCTCTGCAGCATGTTTTGAGCAGGTAAGAGATATCATAACAAATATTGATGGCTCAAAAATAGTAGTTTGTTCTGCAATGGCAGGGGTTACTA

[0534] ACATGTTGGTAGAATTGGTAGAAGCGATAAAAAATGACAAGAGAGATAGTGGTAAAGAAATAATATTGCAATTACAAGAGCATCATGAAAATACA

[0535] GTTGATATATTAAGAATAGATGCTAAGGCTTCAAAAGTATTAAAAGAAAGTATAGCTAGTATAGTAAAGGAGTTATCTTATTTAATAGACAAAGCTTT

[0536] CTCTATAGAATTACAAAATCAGATTGTAACATTCGGAGAGGTTTTATCAACACGTGTTTTTTCTGGTTATCTAGCATCTATAGGTGTTAATAATACAC

[0537] TTTTAATGGCTAAAGACTTTATGTATACCGATAGTTCAGGAAGCCCGGATATTACCACAATACGTGAATACCTAAATGATATTTTACAAACTCATAAA

[0538] GAAGAAAAACTATTTATCACACAAGGATTTATTTGTAAAGATGTACAAGGCAGAGTTAATAATTTAGGACGTGGAGGTAGTGATTATTCAGGAACC

[0539] ATAATTGGAGCGGCATTGCAGGTAGAAGAAGTTCAAATCTGGTCAGATATTGATGGAGTTCATAACAATGATCCAAGGTATGTAGATAATACCAGT

[0540] TCGATTCCATTACTATCTTATCAGGAAGCTTCTGTTTTAGCCAGTTTTGGAGCTAAAGTGCTTCATCCGCAAACGGTTAAGCCTTTGGTAGACCTTA

[0541] AGATACCACTATACTTAAAAAATACTTTTACACCATCTGCTACAGGTACTAAAATAGGCGATAATATATCACCACCAGGAATTAAATCTATTGCAGTA

[0542] AAGGATAATATGGTATGGTTAAAAATAAGAGATATACCCAAAAATGAGAACTATAGCCTATATGATAAAGTATTTACCTTATGTAATGCGTATCAATTT

[0543] CCTGTTATTATGATGGATAATGTAGAAAATGGATTGTTTTTGGTAATAGAAAAAACAGATAAAATATCAGAGTTTTTATATCAGCTTAAACAAGGTTT

[0544] TGACATAGATTTTGATTTACCATATAGTGTAATATGTATTGGAGGAAACTCTATCATGAAAGGCCCAAATAGAGGCCATGTCTTTGGTGCATTGGGA

[0545] GCTATACCAATTAAAATGATTTCTTTTGAGTCGGCAGGCAATTATGTGCTTCTATTAGTAAAGACAGAAGAAAAAATCGATGTAATGCAACAATTAC

[0546] ATCAAAAACTTTTTCATGAGCATGTGGAGTTGGATATGGTATAATTAGAGTTTAATAAGTGAATTGTTTTATTCTGATTCAAAATTGAATAGTGATAT

[0547] CATATCCTTCAAGGTGTTCTTTATGTTTATAAAAGATTGTAGAAATGTAGAAATAAATGTTATCCCTGTTACTGATTAATTTAGAGCGAAAAAATAAC

[0548] ATTTTTATGAGCAATTCAGAAAAATATTTCCTTGTTAATCAAATGTTATTTGTCGGTTAAATAATATCATGATGAGAATGTTTTTGTTTTTTAAAACAC

[0549] TGTTTATTAGTGTTTTAAAAACCCTTGTTAATTGCGTTCTTTTTAAGTGCGGTTTTGCCATAAACATCCTGTTGGTTAAGATTTGTACATTTGAAATG

[0550] GCTTTTAGGAAAAGACGTATTCGTATAAGTTAAATGATATAACTACCCCATTGTTTTTATTGTAAAAAAGATTGATTAAGTGAGATTATTTCTATGCCT

[0551] AATTGATCTCTTTTCAAAAATTATTTTTCAATAATCTTAGTTCATAAAAAACCTTACCAAAAAGGAATTGCGAGTCATATGATGAGCTATATCATCAA

[0552] TTAAATAAACCAATATACCCTTAAAACCTTAGAATATGTGTATTAGTAAACCCGATCAATGGTATGTGCTATATGTAAAGGCACGTCATGAAAGAAA

[0553] AATAGAAAACCTGTTGAAAGAAAATCAAATAGAGACATTTTTACCATTGGTTAAAACAGTTCGAGAATGGAGTGATCGAAAAAAAGAGATAGAA

[0554] ATGCCATTATTTCCTTCCTATATTTTTGTGAAAATTAATTTAAAAAAAGATACGCATAAAGTGTTATCTGTAAAATCGTATTGCTTTTTCATTCGCTTT

[0555] GGAGAAGAATTTGCCAAAGTTCATGAATATGAAATAAACTCAATTAAACTTCTTTTAAATATGAATGTACCTATGAAAGTAGAAAATTATGCACCAT

[0556] TATTAAAAGTAGGCGAATATAGAAGAATAGACAAAGGTGCGCTAAAAGGCTTAAACTGCGAAATCATTAAAGCCGATAATGAAAATAAAGTCGTC

[0557] GTGCGCATTGATTCTATTAAACAAAATATTGTAGCAACAGTACCATCATCCTGCATTTCTGAAGTTGTAGCTTAAATAAATTTTGATAATATAAAAAC

[0558] CCAAAAAATATAAACATCAAGATAGCTCAGTTACTTTCATCATACGTACAATGAGTATGTATGAACGCATAAGGAGGTAACCCTTGTCGTTTTTCAA

[0559] TTTCCACATTTATAAAAATTGCTTTGATAGTAGTAATTCTAGCATTGTCGAGCAATTGTATACTATCGCATAGCTTCATTATCGAAGATTGTGTGATCA

[0560] TCAAATGCATAAATAAATCAAATAACAATCAAAAAACTAAAAAATGAACTCACTTAAAGAATACCATTTATCAGAAACATCAAAAATAGTTTCATC

[0561] TTCGGTACCAGGAGAAAAATCAAAACTCTTACTAGATCTACAAAAAGACATAGAAGGTAGTGTAGTTTCTTATCCGAGAAGTATGCCTATAGCTAT

[0562] AAAAAGAGCTAAAGGAAGTATTATTGAAGATGTAGACGGAAACCAATTTATAGACTTTTTTGGTGGATGTGGTGTTTTAAATGTAGGGCATAGTAA

[0563] TGATTTTGTTTTAGAATATGTAAAAAAACAACAATCAGAATTGATTCATGCCTTAGATTTTCCCACAGAAAATAAACTCTTATTAATTCAAAAAATA

[0564] ATTAACCACTTACCAAAAAGCATTAGAGAAGAATATAAAGTGAGTTTTGGTGGACCCACGGGTTCTGATGCTGTTGAAGCGGCAATCAAGTTAGC

[0565] AAAAATCAAAACTGGTAGAGATGGTATAATCGCTTTTACAGGAGGTTATCACGGTATGACTTCTGGGGCCTTGGCGGTGACTTCGGATACATTTTT

[0566] CAGAAATAATTTAACATCCCTGATGCCAAATGTTCATTTTGCACCTTATAATTACCCTTATAGAAGTACCTCCAATAATAACAATGAGATAGATGGTA

[0567] TAGCTTATTTTAAAAATATACTTACCAACGGTCATTCTGGAATATCTAAACCAGCTGCAATTCTTATAGAACCCATTCAGGGTGAAGGAGGAAATAT

[0568] TGTTCCAGAAGACGGATATTTAGAGGAGTTAGTAAGAATTGCACATGAAAATGATGTACTGGTAATATTTGATGAGATACAGTCTGGTTTTTTTAGA

[0569] ACCGGTAGTTTTTTAGAATTTATGAATACCAGTGCTGTACCAGATATCATCACTTTTTCTAAAGGTTTTGGAGGAATTGGATTTCCAATTTCGGGATT

[0570] AATCTATAAAAAAGAAGTCGAAGCCTGGGGACCTGCCGCACATATAGGGACTTTTAGAGGAAACCAGGTTAGTATAGCTGCAGCAAGAGGAGCC

[0571] TTTGATTTTATTGAAGCCAATAATATAGAAGAACATACGATATTAATTGGTTCCTATTTATTAGAGAAATTACAGGAATTAGAAAGTGAAAATCCGTT

[0572] TATAGGAGAAGTAAGAGGGAGAGGAATGATGATTGGAGTAGAGTTTGTAAAAGATAAAAAGAACAAAGAACCTTTTCCAGAATTTGTAAAGAAA

[0573] TTCAGATCTCAGTGTTTTCAAAGAGGTTTATTGTTTGAAGTAGGAGGGCATTATAACAATGTACTACGACTAGTTCCTCCGTTGGTTACTACCTATA

[0574] ATATTGTAGATGCGGCAATAGCTATTATGAAAGAAGCTATAGCTGTTTGTATGGAAGAATATTTAGAATCTGCTCTCGCCTAAATCCATGTACCACAT

[0575] AACAATGTAACTTATAAAAACAACCCGTTTCCAGAGAGAAAATCTATAAAATAAGTGAGTGCCATTAATGAAATATTCGACACTGTATTTACAAAA

[0576] CTATTTGATACGGGCTCTATTACTAGGTTTGAAAAGATTTAAGAATGCATTCTATAGATATATTTTTTACATCCTTTACAAAACCATTGGATCAGAATC

[0577] TTTTTTCGGAGTATTTGTCCTTGCTACCACCAGACTTAAGGGAAAAAAATTCAAAATACCTTCGTTGGCAAGATAGACATTCGCATTTGTTTGGGC

[0578] GATTGTTATTGATTGAAGCATTAAAGCGTTGTGATATCAACAACATATGGGATCTTATCAAGTATAGTTCACACAAACGACCATATTTAACCTTGGAT

[0579] GGATATGATTTTAACATATCCCATTCTGGAGATTATGTGATTTGTGCAATAGGAAAAAATATCAGGCTTGGTATTGATATTGAAGAACACCGAGAGA

[0580] GAAACCTAAAAGATTTCAGGAACTTAATGACTCTGGATCAATGGGATGAAATTAACAGTGCGAGTTGTTCGTTAAATACGTTTTATAAATATTGGA

[0581] CGATTAAGGAAAGCGTTATTAAGGCAGATGGTCGCGGGTTTTATATTCCTCTAAATGAGCTTGAAGTGACCAATAACACGGTTTGTTTTGAGGATA

[0582] AATTATGGTTTGTGCAGGATTTTGTATTTACCAAAGGGTATAGTGTAGCAATCGCAACCAATGAGTTATCAACATTTAACATGCATGCTATAAATTTT

[0583] TATGACTCTTGTATAGTATAGGTCGATGTATAGCATTTACGATTTCAATCTTTCGGTAATTTCAAAACATAGATGGTTAAACTAATCCTATTCTTCAAC

[0584] AAGATCTTAGGTCGAATACCTTGTGGGTTTACCCCAAGGATATTTACTCTTTTTTTTTCGAATTACTAATCATACCAACACTTAAAACCGTTCTAAAT

[0585] CCTAGATGCTCAAGCGAAGAATCTGTGCTAGTAGCCATTCGTGCACTATTACGATAACTGGCACAATAAGAATCGCTACATAAAAAAGAACCGCCT

[0586] TTTATGACTTTTTCTCGTAAAGCAGGATTATTAGGATTATACGGTTTTTCGGCACCTTTAGGATTATGAGTTAATCCTTGTTGTTTTAATTGATTGTAA

[0587] TACGTTGTATTGTACCAATCTTGTGTCCATTCCCAAACGTTCCCTGCCATGTCATAGAGATGATATGAATTAGGCGGATAGGAACGAACAGGGGCTT

[0588] TATTTTCATAACCATCTGCTTTGGTATTTTGT

Claims

1. A dual-target antimicrobial lipopeptide, characterized in that: The lipopeptide can bind to dual targets on Gram-positive bacteria or Gram-negative bacteria.

2. The antimicrobial lipopeptide according to claim 1, characterized in that The lipopeptide is obtained through a structure prediction chemical synthesis strategy based on a non-ribosomal peptide synthesis gene cluster encoded by Aquimarina sp. EL43. The non-ribosomal peptide synthesis gene cluster is numbered No. PQ867817 in GenBank, and the sequence is shown in SEQ ID NO.

1.

3. The antimicrobial lipopeptide according to claim 1, characterized in that The dual targets on the Gram-positive bacteria are cardiolipin (CL) and phosphatidylglycerol (PG), and the dual targets on the Gram-negative bacteria are lipopolysaccharide (LPS) and phosphatidylethanolamine (PE).

4. The antimicrobial lipopeptide according to claim 1, characterized in that The antimicrobial lipopeptide is a linear lipopeptide aquicidine L, i.e., Aquicidine L, with a chemical formula of C 70 H 116 N 16 O 16 , the chemical structure is shown below:

5. The lipopeptide according to claim 1, characterized in that The lipopeptide is a cyclic lipopeptide aquicidine C4, i.e., Aquicidine C4, with a chemical formula of C 70 H 114 N 16 O 15 , the chemical structure is shown below:

6. A method for preparing a dual-target antimicrobial lipopeptide according to any one of claims 1 to 5, characterized in that According to the law of non-ribosomal peptide synthesis, that is, using the composition of amino acids in the conserved position of the active center site of the adenylation domain enzyme, the substrate it recognizes is directly predicted, thereby deducing the linear polypeptide sequence encoded by the non-ribosomal peptide synthesis gene cluster encoded by Aquimarina sp. EL43, and then adding the myristic acid fatty acid side chain, the dual-target antimicrobial lipopeptide is obtained by using a polypeptide solid phase synthesis strategy.

7. The preparation method according to claim 6, characterized in that: The preparation method comprises the following steps: Step 1, loading the amino acid onto 2-chlorotrityl resin and swelling it in dichloromethane (DCM), followed by draining the solvent and washing it with N,N-dimethylformamide (DMF); Step 2: Mix the Fmoc-protected amino acid with 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) in DMF, and gradually couple each amino acid. The coupling reaction is carried out for 1 hour, with occasional stirring and washing with DMF. Step 3, coupling the remaining amino acids; Step 4: Complete the final cleavage of the linear or cyclic peptide to remove the N-Fmoc protecting group. Dissolve each peptide in a cleavage mixture (95% trifluoroacetic acid, 2.5% triisopropylsilane, and 2.5% water) and react for 1.5 hours. Then, add a 1:1 precooled ether:hexane mixture and let stand at -20°C for 10 minutes to precipitate the linear or cyclic peptide. Step 5: Collect the polypeptide precipitate, redissolve it in methanol, and dry it under vacuum.

8. The preparation method according to claim 7, characterized in that: When preparing the linear lipopeptide helobactin L described in claim 4, the synthesis is started from the last amino acid, and step 3 specifically includes: Fmoc deprotection is carried out using a 20% piperidine DMF solution for 7 minutes, repeated twice, and after the resin is washed with DMF, it is coupled with the next N-Fmoc protected amino acid.

9. The preparation method according to claim 7, characterized in that: When preparing the cyclic lipopeptide helobactin C4 according to claim 5, the synthesis starts from the fifth amino acid.

10. The preparation method according to claim 9, characterized in that: Step 3 specifically comprises: mixing the resin with benzoyl chloride, DIPEA, the last amino acid of the peptide and 4-dimethylaminopyridine (DMAP) in DCM and rotating slowly for 72 hours, and after the ester bond is formed, coupling the remaining amino acids according to the method of step 2; The resin was removed by treatment with 20% hexafluoroisopropanol, i.e., HFIP, in DCM for 2 h; The soluble peptide was then cyclized using (7-azabenzotriazole-1-oxy)tripyrrolylphosphonium hexafluorophosphate, PYAOP, and DIPEA in DMF and extracted with 1% formic acid in DCM and H2O. The cyclized peptide was dissolved in methanol and dried under vacuum overnight.

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