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É DADO O SEGUINTE ALINHAMENTO: CLUSTAL 2.1 multiple sequence alignment 1 -----------ATT-TCCTTGCCTTAAA----TCAATCAA----CCGTACTCACA--CTG 38 5 ---------GGACTATCCTCTCCTCAAAGATCCCCACC------CCGTATTCAAT--TCC 43 4 CCCACTTTGGAATTTTCTTTACCTTA------CCTAAGAA----GGGTATTAATT--TGA 48 2 CTCTCCTCGTGGTCGCCAGCACTGCCGTG---CCGAGCCGGAGGCCGAGCTCGGCAGCTG 57 3 ----------GGCCCTCA-CTCTCCC------TCGGTCC----------CTCGTC----- 28 * * * * 1 TTCTTCTTCTT-----------CTAACAACTTCTGC--------AATTTGTTCTCA---- 75 5 CTCTTCTCCTT-----------CTCTGAATTGCT------------TTCCATCTC----- 75 4 TTCTTGTGGGAAGGAAGAAGGATCAAGAATGGCG----------ATTTCGATTTCT---- 94 2 CCCGCCTCGGTGTGTCGC--CGCCGCACCCCACCCACGTAGCAAAAGCCGTCTTCCCGTA 115 3 TCCACCTTCGC---TCGCG-CGCCG-GCGCCGCTCAC--------ACCCGATTTCC---A 72 * * * ** 1 -TTTATG--TTCAG-----ACAGCTTCTCAGCAGC-----------TAATCTCT-CAAAA 115 5 -TTTGTG--TTTTC-----TCCATTTTTT----GT-----------T--TCTTT-CTGGA 109 4 -GCTATGAGTTTTGGAACCTCAGTTTCTTCA---------------TATTCTTG-TTTTA 137 2 CCGTAGGCAGCGGCA---AACAGCGCCCCCTCATCACTCGGTCTCGTTCCCTCCTCTCCA 172 3 CCTCACAAAACC-CA---CACGGAACCCCGGCACC--------------CCTCC------ 108 * ** 1 ATAATAAGAAAATGGCAGCAG------CAGGTCTCTCTTCTTCTTCATGTACT-GTATTC 168 5 GAGAT---AGAATGGC--CA-------TGAGTATCTCACCG-------GCAGC-ACATCC 149 4 GAGCT---AGGA--GTTTTGA------GAAGT----CATCA---------GTT-TTATGC 172 2 ACTTCACTCGCGTCGCGCCCCTCCACTGCACCAG--CGTCATG-----GCGGTGGCCTCG 225 3 ACTCCACCAG----GCGCC----------AGCGT--CGTCATG-----GCGGTAGCCTCG 147 * * * * 1 ACACTCTCCTCATCCTTTAAAACCAGAA-GACA-TCTCACTAAAACCCCACAAAACCCTT 226 5 A-ACCTCAATCTCCATTAGAAATTGTAA-AACCCTTTCCCCTTTCCCCCGCCAAA----A 203 4 A----------ATTCCCAGAACCCAT---GTCGGTTTAATTCTGTTTTTCCGA------- 212 2 ACCTCGCCGCTGTCCGCCAAGCCCGCCACGGCCCCTTCGCCGCCCGCTCCCGGA------ 279 3 ACGTCGCCCCTTTCCGCCAAGCCCACCACGGCCACCTCGCCGCCCACTCCCGCA------ 201 * * * * * 1 TGCTTTTCAAACGCAACAACAGCAGCAACCGCACTTCACATCCCTTTCTT----CCTTCC 282 5 TG-TTTTGAGATACAATCCC---------CTCTCTCCTCGTCGATCTAAA----GTCTCT 249 4 ---TTCGGAAATCTGATGGG------------GCTTCACGGTGTTCT-------GTTTCT 250 2 ---TCCGGGCTCCTCGCTCTCGG------CGTTCGCCGCGCCCCCGCCACTGCCGCGTGG 330 3 ---TTCGGCTTCCTCGCTCCCCG------CTCCCGCCGCGGCCGCGCCAC---CGCGTGG 249 * * * * * 1 AGAAAGTT--GTGTAGAGTTCAAGCAACAATTTTGAGAGAAGATGAAGAGAAGAAAGTTG 340 5 GGGAAGCT--GTTCCGAGTTCAAGCAACAGTTTTGCGAGAAAATGATGAG---AAAGTGG 304 4 AGGAAATC--ATGTAGAGTTCGAGCAACGTTGTTACAAGAGAATGAAGAA---GAAGTGG 305 2 AGGAGGCTCCGCGTGGAGG-CGATCAG-GACGCAGCGA------------ACGGAGGTGC 376 3 AGGAGTCTCCTCGTGGAGG-CGATCCG-GACGCAGCGGGAGAAACAGGGGACGGAGGGGC 307 * * *** * * * * * 1 TTGTTGAGGAATCATTTCAACCAAAG---------------ACTTTCACTCATGAGCC-- 383 5 TGGTAGAGGAAACATTTCAACCCAAA---------------ACTTCTACAAATGAGGAGA 349 4 TTGTGGAGAAATCTTTTGCACCTAAG---------------AGTTTTCCTGATAACGTGG 350 2 CCGTCGAGGAGTCCGCCCCCGCCAGGGACGCCGCCGCTGCCGCGCCCCTGGACGGAAACG 436 3 ACTTCGAGGAGTCCGCC---------------------GCCGCGCCCCTGGACGGAGTCG 346 * *** * * * 1 -------------TGTTCGGGGCTCACCACAATCTTCATCGCCTGGTGGATTAGAGACTT 430 5 GAAAAGGAGGAGATGGT-GAGCCCCAGGATGA--TTCCTCATCAGGTACTCTGGAGAAAT 406 4 GAGGGGGAAGTAATGGG-AAGCCACCAGATGA--TTCATCCTCTAACGGTCTAGAGAAAT 407 2 GAGCCGGAGCGGACG-----GCTCCGTGGT----TCCTTCCTCGGACG------ACAGCT 481 3 GAGTCGGAGCCGATG-----ACCCCGTGGT----TCCCTCCTCGGACGC---GAGCGACT 394 * * * * ** * * 1 GGGCTATCAAGCTTGAGCAATCTGTCAATGTCTTTCTCACTGATTCGGTGATAAAAATTC 490 5 GGGTTATCAAGCTTGAACAATCTATCAATATCTTTCTCACGGATTCAGTGATAAAGGTTC 466 4 GGGTTATAAAGCTTGAGCAGTCTGTAAATATCTTACTCACGGATTCAGTGATAAAGATTC 467 2 GGGTTGTCAAGCTCGAGCAGTCGTTCAACATTTTCGCCACGGATTCGGTGATTATGGTAC 541 3 GGGTGGTCAAGCTCGAGCAGTCCTTCAACATTTTCGCCACGGATTCGGTGATAATGGTAC 454 *** * ***** ** ** ** * ** * ** *** ***** ***** * * * 1 TTGATACTCTTTATCACGACCGCGATTATGCTAGGTTCTTTGTTTTGGAAACTATTGCTA 550 5 TAGATTCTTTGTACCATGACCGAGATTATGCAAGGTTCTTCGTGTTGGAAACTATTGCAA 526 4 TTGACACTTTGTATCACAACCGAAACTATGCGAGGTTTTTTGTTCTGGAAACAATTGCAA 527 2 TCAAGGGCGTGTACGGTGATCGGTACTACGCCAGGTTCTTTGCGCTGGAGACGATTGCGA 601 3 TCAAGGGCGTGTACCGTGATCGGTACTACGCCAGGTTCTTTGCGCTGGAGACGATTGCCA 514 * * * ** * ** * ** ** ***** ** * **** ** ***** * 1 GAGTTCCTTATTTTGCCTTTATATCTGTTCTTCACATGTATGAGAGTTTTGGTTGGTGGA 610 5 GAGTTCCTTATTTTGCCTTTATATCTGTCCTTCATATGTATGAGAGTTTTGGTTGGTGGA 586 4 GGGTTCCTTATTTTGCATTTATATCGGTTCTTCACATGTATGAGAGCTTTGGCTGGTGGA 587 2 GGGTGCCGTACTTCGCATTCATATCGGTGCTTCACTTGTATGCGACCTTTGGATGGTGGA 661 3 GGGTGCCGTATTTCGCGTTCATATCGGTGCTTCACATGTATTCGACCTTTGGCTGGTGGA 574 * ** ** ** ** ** ** ***** ** ***** ***** ** ***** ******* 1 GAAGAGCTGATTATCTCAAAGTGCATTTTGCTGAGAGCTGGAATGAGATGCATCACTTGC 670 5 GAAGAGCGGATTATCTCAAAGTACATTTTGCTGAGAGCTGGAATGAGATGCACCATTTGC 646 4 GAAGGGCAGATTATATGAAAGTGCATTTTGCTGAAAGCTGGAATGAGATGCACCATTTGC 647 2 GACGAGCTGATTACATAAAGGTTCACTTTGCGCAGAGCTGGAACGAGTTCCATCACCTCT 721 3 GACGAGCGGATTATATAAAGGTTCACTTTGCGCAAAGCTGGAACGAGTTCCATCACCTCT 634 ** * ** ***** * ** ** ** ***** * ******** *** * ** ** * 1 TTATCATGGAAGAATTGGGTGGTAATTCTTGGTGGTTTGACCGGCTTCTTGCTCAAGTTA 730 5 TTATCATGGAAGAATTGGGGGGCAATGCTTGGTGGTTTGATCGGTTTCTTTCCCAACATA 706 4 TCATTATGGAAGAATTAGGGGGAAATGCTTGGTGGTTTGATCGATTTCTTGCACAACATA 707 2 TGATCATGGAAGAATTGGGTGGCGACTCTTTGTGGTTTGACTGTTTTCTTGCTCGGTTTA 781 3 TGATCATGGAAGAATTGGGTGGCAACTCTTTATGGATTGACTGTTTCCTTGCTCGGTTTA 694 * ** *********** ** ** * *** *** **** * * *** * * ** 1 TAGCAACCTCTTATTATTTCATGACAGTCTTAATGTATGCATTGAGCCCAAGAATGGCAT 790 5 TAGCCGTCGTGTACTATTTTATGACAGTTCTCATGTATGCAATAAGCCCAAGAATGGCTT 766 4 TAGCTATATTCTATTATTTCATGACAGTCTTGATGTATGCTTTGAGCCCGAGAATGGCAT 767 2 TGGCATTCTTTTACTACTTCATGACTGTTGCAATGTACATGCTGAGCCCACGAATGGCAT 841 3 TGGCGTTTTTTTACTACTTCGTGACTGTTGCGATGTACATGCTGAGCCCAAGAATGGCAT 754 * ** ** ** ** **** ** ***** * ***** ******* * 1 ATCACTTCTCTGAATGTGTTGAGAGCCATGCATTTGCAACTTATGACAAATTTATCAAGG 850 5 ATCACTTTTCTGAATGCGTGGAAAGCCATGCATTTTCAACTTATGACAAATTTATCAAAG 826 4 ATCATTTCTCTGAATGTGTGGAGAGCCATGCATACGAGACTTACGATAAATTCATCAAGG 827 2 ATCACTTTTCCGAATGTGTGGAGAGACATGCATATTCCACCTATGATGAGTTCCTCAAGC 901 3 ATCACTTTTCTGAATGTGTGGAGAGACATGCATATTCCACCTATGACGAGTTCCTCAAGC 814 **** ** ** ***** ** ** ** ******* ** ** ** * ** **** 1 CCCAAGGAGATGATTTGAAAAAATTGCCTGCACCTGAGGTTGCTGTAAAATATTATACCG 910 5 CACAAGGAGAGGAGTTGAAAAAATTACCTGCTCCTGAAGTTGCTATAAAATACTACACTG886 4 ATCAAGGAGAGGAATTGAAGAATTTGCCCGCTCCAAAGATTGCAGTGGACTACTACACGG 887 2 TCCATGAAGAGGAATTGAAAAGACTACCAGCTCCAGAGGCAGCATTGAACTATTACATGA 961 3 TCCATGAAGAGGAATTGAAAAGACTACCAGCTCCAGAGGCAGCATTAGAATATTACCTGA 874 ** * *** ** ***** * * ** ** ** * ** * * ** ** 1 AGGGTGATTTGTACTTGTTTGATGAATTTCAAACTTCCAGAGCTCCCCATTCTCGTAGGC 970 5 GTGGAGACTTGTACTTGTTTGATGAGTTTCAAACTTCCAGAGCCCCCAATACACG----- 941 4 GAGGTGACTTATATTTATTTGATGAGTTTCAAACTTCACGAGAGCCTAATACTCGAAGAC 947 2 ATGAGGACCTTTACTTATTCGATGAGTTTCAGGCATCAAGAACTCCAGGTTCTAGGAGGC 1021 3 ATGAGGACCTTTACTTATTCGATGAGTTTCAGGCATCAAGAAGTCCAGGTTCTAGGAGGC 934 * ** * ** ** ** ***** ***** * ** ** ** * * * 1 CAAAAATAGAGAATTTGTATGATGTATTTCTGAACGTCAGAGATGATGAGGCTGAACATT 1030 5 ------------------------------------------------------------ 4 CAAAAATAGATAATCTCTATGACGTATTCATGAACATTAGAGATGACGAAGCAGAGCATT 1007 2 CTAAAATAGATAACTTATACGATGTATTCGTTAATATACGAGAAGATGAGGCAGAGCACT 1081 3 CTAAAATAGATAACTTATATGATGTATTTGTCAATATACGAGATGACGAGGCAGAACACT 994 1 GTAAGACCATGAAGGCCTGCCAAACACATGGAAATCTCCGCTCTCCACATTCATATCCAG 1090 5 ------------------------------------------------------------ 4 GTAAAACGATGAAAGCCTGTCAAACTCACGGGAGCCTTCGTTCTCCACACACAGATCCAT 1067 2 GCAAGACAATGAAGACCTGTCAAACACATGGAAATCTTCGTTCTCCTCATTCAA---CGC 1138 3 GCAAGACAATGAAGACCTGTCAAACACACGGAAATCTTCGTTCTCCTCACTCAA---CGC 1051 1 AGGATGCTTTTGAAGATGATACTGGCTGTGATCTTCCTCAAGCAGATTGTGAAGGTATTG 1150 5 ------------------------------------------------------------ 4 GCGATGATTCTGAAGATGATACAGGGTGTTCCGTACCTCAAGCTGATTGTATAGGTATCG 1127 2 CGAACTGCTTAGAAGATGATACGGAATGTGTAATACCTGAAAACGACTGTGAAGGTATTG 1198 3 AGAACTGCTTAGAAGCTGATACGGAATGTGTAATACCTGAAAACGATTGTGAAGGTATTG 1111 1 TTGATTGTATAAAGAAATCTGTAACATCTCCTCCATCAAAGCAAAATATTTGAAGGAGAA 1210 5 ------------------------------------------------------------ 4 TGGATTGTATAAAGAAGTCAGTCACCGATACTCAAGTAAC-CAAAAGGTAGGAAAAGGAA 1186 2 TGGACTGTGTCAAAAAGTCCCTTAC---------------------------AAAGTAAA 1231 3 TGGACTGTGTCAAAAAGTCCCTTAC---------------------------AAAGTAAA 1144 1 CAA---GGAAACAGTATACATTT-----------CAGGTGATGGA--------------- 1241 5 ------------------------------------------------------------ 4 AAACGCGGACAAACTATACTTGTATATACTAGTATAGACAAAAAAAAAAAAATACAAAGA 1246 2 TAG---------ACTATATTCTTT----------AATGTGTTTGA--------------C 1258 3 TAG---------ACTATATTCTTT----------CGTGT--------------------- 1164 1 --TAGTGAGATATTGTAT-----------ACATAGAAA-------GAAACATTACAA--- 1278 5 ------------------------------------------------------------ 4 TATAGGTACATGTTGTATCTTTTTGTCTGAGGTAGAGATGTTTTGGAAAGTTTTCAAGGC 1306 2 TGTTTTCAGGTGTCGTA------------GTATACAAAG----TATAAAATTTTGATG-- 1300 3 -GTTTCCAGGCGTCATA------------GTATACAAAG----TACAAAATTTTTATG-- 1205 1 -ATTTACATAATTTACAT-------------------------GCCTTCAAGCTAAAAAT 1312 5 ------------------------------------------------------------ 4 AAAATACATAGTTTACCTCATGTAAGCTCTCCTTACTAAGATTGTTTTTATATTTAGAAT 1366 2 -ATATCCTCTCT------------------------------------------------ 1311 3 -ATATCCTCTTTTTTCCT---------------------ATGTACCTTTTTAAGTAAAAT 1243 1 ------------------------------------------------------------ 5 ------------------------------------------------------------ 4 TTGTACCCGAAACCTGTTATAGGTGGATGGATCTTGACAACTTTTGGATATATCGACTCC 1426 2 ------------------------------------------------------------ 3 CTGGGTCATATGTAAGAGAAGCCCAGTTTAGAACTTTAGATACAATTATGCAAC------ 1297 1 ------------------------------------- 5 ------------------------------------- 4 ATGAGCTTTTTACACACGCAAAAAAAAAAAAAAAAAA 1463 2 ------------------------------------- 3 ------------------------------------- 1 ) Obter as sequências no formato fasta. Para obter a sequência no formato FASTA copiar toda a sequência com o nome CLUSTAL e levar para o site https://www-bimas.cit.nih.gov/molbio/readseq/ sequências cheias de gap ir na ferramenta reverso complemento fazer reverso complemento copiar o resultado e fazer novamente o reverso complemento. >1 ATTTCCTTGCCTTAAATCAATCAACCGTACTCACACTGTTCTTCTTCTTCTAACAACTTCTGCAATTTGTTCTCATTTATGTTCAGACAGCTTCTCAGCAGCTAATCTCTCAAAAATAATAAGAAAATGGCAGCAGCAGGTCTCTCTTCTTCTTCATGTACTGTATTCACACTCTCCTCATCCTTTAAAACCAGAAGACATCTCACTAAAACCCCACAAAACCCTTTGCTTTTCAAACGCAACAACAGCAGCAACCGCACTTCACATCCCTTTCTTCCTTCCAGAAAGTTGTGTAGAGTTCAAGCAACAATTTTGAGAGAAGATGAAGAGAAGAAAGTTGTTGTTGAGGAATCATTTCAACCAAAGACTTTCACTCATGAGCCTGTTCGGGGCTCACCACAATCTTCATCGCCTGGTGGATTAGAGACTTGGGCTATCAAGCTTGAGCAATCTGTCAATGTCTTTCTCACTGATTCGGTGATAAAAATTCTTGATACTCTTTATCACGACCGCGATTATGCTAGGTTCTTTGTTTTGGAAACTATTGCTAGAGTTCCTTATTTTGCCTTTATATCTGTTCTTCACATGTATGAGAGTTTTGGTTGGTGGAGAAGAGCTGATTATCTCAAAGTGCATTTTGCTGAGAGCTGGAATGAGATGCATCACTTGCTTATCATGGAAGAATTGGGTGGTAATTCTTGGTGGTTTGACCGGCTTCTTGCTCAAGTTATAGCAACCTCTTATTATTTCATGACAGTCTTAATGTATGCATTGAGCCCAAGAATGGCATATCACTTCTCTGAATGTGTTGAGAGCCATGCATTTGCAACTTATGACAAATTTATCAAGGCCCAAGGAGATGATTTGAAAAAATTGCCTGCACCTGAGGTTGCTGTAAAATATTATACCGAGGGTGATTTGTACTTGTTTGATGAATTTCAAACTTCCAGAGCTCCCCATTCTCGTAGGCCAAAAATAGAGAATTTGTATGATGTATTTCTGAACGTCAGAGATGATGAGGCTGAACATTGTAAGACCATGAAGGCCTGCCAAACACATGGAAATCTCCGCTCTCCACATTCATATCCAGAGGATGCTTTTGAAGATGATACTGGCTGTGATCTTCCTCAAGCAGATTGTGAAGGTATTGTTGATTGTATAAAGAAATCTGTAACATCTCCTCCATCAAAGCAAAATATTTGAAGGAGAACAAGGAAACAGTATACATTTCAGGTGATGGATAGTGAGATATTGTATACATAGAAAGAAACATTACAAATTTACATAATTTACATGCCTTCAAGCTAAAAAT >2 CTCTCCTCGTGGTCGCCAGCACTGCCGTGCCGAGCCGGAGGCCGAGCTCGGCAGCTGCCCGCCTCGGTGTGTCGCCGCCGCACCCCACCCACGTAGCAAAAGCCGTCTTCCCGTACCGTAGGCAGCGGCAAACAGCGCCCCCTCATCACTCGGTCTCGTTCCCTCCTCTCCAACTTCACTCGCGTCGCGCCCCTCCACTGCACCAGCGTCATGGCGGTGGCCTCGACCTCGCCGCTGTCCGCCAAGCCCGCCACGGCCCCTTCGCCGCCCGCTCCCGGATCCGGGCTCCTCGCTCTCGGCGTTCGCCGCGCCCCCGCCACTGCCGCGTGGAGGAGGCTCCGCGTGGAGGCGATCAGGACGCAGCGAACGGAGGTGCCCGTCGAGGAGTCCGCCCCCGCCAGGGACGCCGCCGCTGCCGCGCCCCTGGACGGAAACGGAGCCGGAGCGGACGGCTCCGTGGTTCCTTCCTCGGACGACAGCTGGGTTGTCAAGCTCGAGCAGTCGTTCAACATTTTCGCCACGGATTCGGTGATTATGGTACTCAAGGGCGTGTACGGTGATCGGTACTACGCCAGGTTCTTTGCGCTGGAGACGATTGCGAGGGTGCCGTACTTCGCATTCATATCGGTGCTTCACTTGTATGCGACCTTTGGATGGTGGAGACGAGCTGATTACATAAAGGTTCACTTTGCGCAGAGCTGGAACGAGTTCCATCACCTCTTGATCATGGAAGAATTGGGTGGCGACTCTTTGTGGTTTGACTGTTTTCTTGCTCGGTTTATGGCATTCTTTTACTACTTCATGACTGTTGCAATGTACATGCTGAGCCCACGAATGGCATATCACTTTTCCGAATGTGTGGAGAGACATGCATATTCCACCTATGATGAGTTCCTCAAGCTCCATGAAGAGGAATTGAAAAGACTACCAGCTCCAGAGGCAGCATTGAACTATTACATGAATGAGGACCTTTACTTATTCGATGAGTTTCAGGCATCAAGAACTCCAGGTTCTAGGAGG CCTAAAATAGATAACTTATACGATGTATTCGTTAATATACGAGAAGATGAGGCAGAGCACTGCAAGACAATGAAGACCTGTCAAACACATGGAAATCTTCGTTCTCCTCATTCAACGCCGAACTGCTTAGAAGATGATACGGAATGTGTAATACCTGAAAACGACTGTGAAGGTATTGTGGACTGTGTCAAAAAGTCCCTTACAAAGTAAATAGACTATATTCTTTAATGTGTTTGACTGTTTTCAGGTGTCGTAGTATACAAAGTATAAAATTTTGATGATATCCTCTCT>3 GGCCCTCACTCTCCCTCGGTCCCTCGTCTCCACCTTCGCTCGCGCGCCGGCGCCGCTCACACCCGATTTCCACCTCACAAAACCCACACGGAACCCCGGCACCCCTCCACTCCACCAGGCGCCAGCGTCGTCATGGCGGTAGCCTCGACGTCGCCCCTTTCCGCCAAGCCCACCACGGCCACCTCGCCGCCCACTCCCGCATTCGGCTTCCTCGCTCCCCGCTCCCGCCGCGGCCGCGCCACCGCGTGGAGGAGTCTCCTCGTGGAGGCGATCCGGACGCAGCGGGAGAAACAGGGGACGGAGGGGCACTTCGAGGAGTCCGCCGCCGCGCCCCTGGACGGAGTCGGAGTCGGAGCCGATGACCCCGTGGTTCCCTCCTCGGACGCGAGCGACTGGGTGGTCAAGCTCGAGCAGTCCTTCAACATTTTCGCCACGGATTCGGTGATAATGGTACTCAAGGGCGTGTACCGTGATCGGTACTACGCCAGGTTCTTTGCGCTGGAGACGATTGCCAGGGTGCCGTATTTCGCGTTCATATCGGTGCTTCACATGTATTCGACCTTTGGCTGGTGGAGACGAGCGGATTATATAAAGGTTCACTTTGCGCAAAGCTGGAACGAGTTCCATCACCTCTTGATCATGGAAGAATTGGGTGGCAACTCTTTATGGATTGACTGTTTCCTTGCTCGGTTTATGGCGTTTTTTTACTACTTCGTGACTGTTGCGATGTACATGCTGAGCCCAAGAATGGCATATCACTTTTCTGAATGTGTGGAGAGACATGCATATTCCACCTATGACGAGTTCCTCAAGCTCCATGAAGAGGAATTGAAAAGACTACCAGCTCCAGAGGCAGCATTAGAATATTACCTGAATGAGGACCTTTACTTATTCGATGAGTTTCAGGCATCAAGAAGTCCAGGTTCTAGGAGGCCTAAAATAGATAACTTATATGATGTATTTGTCAATATACGAGATGACGAGGCAGAACACTGCAAGACAATGAAGACCTGTCAAACACACGGAAATCTTCGTTCTCCTCACTCAACGCAGAACTGCTTAGAAGCTGATACGGAATGTGTAATACCTGAAAACGATTGTGAAGGTATTGTGGACTGTGTCAAAAAGTCCCTTACAAAGTAAATAGACTATATTCTTTCGTGTGTTTCCAGGCGTCATAGTATACAAAGTACAAAATTTTTATGATATCCTCTTTTTTCCTATGTACCTTTTTAAGTAAAATCTGGGTCATATGTAAGAGAAGCCCAGTTTAGAACTTTAGATACAATTATGCAAC >4 CCCACTTTGGAATTTTCTTTACCTTACCTAAGAAGGGTATTAATTTGATTCTTGTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACCTCAGTTTCTTCATATTCTTGTTTTAGAGCTAGGAGTTTTGAGAAGTCATCAGTTTTATGCAATTCCCAGAACCCATGTCGGTTTAATTCTGTTTTTCCGATTCGGAAATCTGATGGGGCTTCACGGTGTTCTGTTTCTAGGAAATCATGTAGAGTTCGAGCAACGTTGTTACAAGAGAATGAAGAAGAAGTGGTTGTGGAGAAATCTTTTGCACCTAAGAGTTTTCCTGATAACGTGGGAGGGGGAAGTAATGGGAAGCCACCAGATGATTCATCCTCTAACGGTCTAGAGAAATGGGTTATAAAGCTTGAGCAGTCTGTAAATATCTTACTCACGGATTCAGTGATAAAGATTCTTGACACTTTGTATCACAACCGAAACTATGCGAGGTTTTTTGTTCTGGAAACAATTGCAAGGGTTCCTTATTTTGCATTTATATCGGTTCTTCACATGTATGAGAGCTTTGGCTGGTGGAGAAGGGCAGATTATATGAAAGTGCATTTTGCTGAAAGCTGGAATGAGATGCACCATTTGCTCATTATGGAAGAATTAGGGGGAAATGCTTGGTGGTTTGATCGATTTCTTGCACAACATATAGCTATATTCTATTATTTCATGACAGTCTTGATGTATGCTTTGAGCCCGAGAATGGCATATCATTTCTCTGAATGTGTGGAGAGCCATGCATACGAGACTTACGATAAATTCATCAAGGATCAAGGAGAGGAATTGAAGAATTTGCCCGCTCCAAAGATTGCAGTGGACTACTACACGGGAGGTGACTTATATTTATTTGATGAGTTTCAAACTTCACGAGAGCCTAATACTCGAAGACCAAAAATAGATAATCTCTATGACGTATTCATGAACATTAGAGATGACGAAGCAGAGCATTGTAAAACGATGAA AGCCTGTCAAACTCACGGGAGCCTTCGTTCTCCACACACAGATCCATGCGATGATTCTGAAGATGATACAGGGTGTTCCGTACCTCAAGCTGATTGTATAGGTATCGTGGATTGTATAAAGAAGTCAGTCACCGATACTCAAGTAACCAAAAGGTAGGAAAAGGAAAAACGCGGACAAACTATACTTGTATATACTAGTATAGACAAAAAAAAAAAAATACAAAGATATAGGTACATGTTGTATCTTTTTGTCTGAGGTAGAGATGTTTTGGAAAGTTTTCAAGGCAAAATACATAGTTTACCTCATGTAAGCTCTCCTTACTAAGATTGTTTTTATATTTAGAATTTGTACCCGAAACCTGTTATAGGTGGATGGATCTTGACAACTTTTGGATATATCGACTCCATGAGCTTTTTACACACGCAAAAAAAAAAAAAAAAAA >5 GGACTATCCTCTCCTCAAAGATCCCCACCCCGTATTCAATTCCCTCTTCTCCTTCTCTGAATTGCTTTCCATCTCTTTGTGTTTTCTCCATTTTTTGTTTCTTTCTGGAGAGATAGAATGGCCATGAGTATCTCACCGGCAGCACATCCAACCTCAATCTCCATTAGAAATTGTAAAACCCTTTCCCCTTTCCCCCGCCAAAATGTTTTGAGATACAATCCCCTCTCTCCTCGTCGATCTAAAGTCTCTGGGAAGCTGTTCCGAGTTCAAGCAACAGTTTTGCGAGAAAATGATGAGAAAGTGGTGGTAGAGGAAACATTTCAACCCAAAACTTCTACAAATGAGGAGAGAAAAGGAGGAGATGGTGAGCCCCAGGATGATTCCTCATCAGGTACTCTGGAGAAATGGGTTATCAAGCTTGAACAATCTATCAATATCTTTCTCACGGATTCAGTGATAAAGGTTCTAGATTCTTTGTACCATGACCGAGATTATGCAAGGTTCTTCGTGTTGGAAACTATTGCAAGAGTTCCTTATTTTGCCTTTATATCTGTCCTTCATATGTATGAGAGTTTTGGTTGGTGGAGAAGAGCGGATTATCTCAAAGTACATTTTGCTGAGAGCTGGAATGAGATGCACCATTTGCTTATCATGGAAGAATTGGGGGGCAATGCTTGGTGGTTTGATCGGTTTCTTTCCCAACATATAGCCGTCGTGTACTATTTTATGACAGTTCTCATGTATGCAATAAGCCCAAGAATGGCTTATCACTTTTCTGAATGCGTGGAAAGCCATGCATTTTCAACTTATGACAAATTTATCAAAGCACAAGGAGAGGAGTTGAAAAAATTACCTGCTCCTGAAGTTGCTATAAAATACTACACTGGTGGAGACTTGTACTTGTTTGATGAGTTTCAAACTTCCAGAGCCCCCAATACACG 2) Identificar cada uma das sequencias: Função (se existente) e organismo Para essa questão vamos usar o BLAST, utilize os seguintes bancos de dados Refseq - RNA, EST e TSA não selecione o organismo (até porque não sabemos qual é o organismo). Com Refseq – RNA vamos encontrar a função e com os demais EST e TSA o organismo, realize um MEGABLAST. Sequência 1 Organismo: Populus trichocarpa, Função: Ubiquinonona oxidase 4 Sequência 2 Organismo: Zea mays, Função: Ubiquinonona oxidase 4 Sequência 3 Organismo: Sorghum bicolor , Função: Ubiquinonona oxidase 4 Sequência 4 Organismo: Solanum lycopersicum, Função: Ubiquinonona oxidase 4 Sequência 5 Organismo:, Função: Euphorbia esula, Função: Ubiquinonona oxidase 4 3) Em caso de corresponder a cDNA ou gene, informar a sequência de aminoácidos deduzidos Vamos nos guiar pelo Refseq- RNA. Vamos ter o cDNA observe se a sequência aumenta aumenta, caso não fazer o REVERSO COMPLEMENTO MUITA ATENÇÃO!!!! Utilizar o Expasy translate Tool pegar cada sequência e levar para a ferramenta. Coloque as sequências como no exemplo a seguir: >Seq1, ATENÇÃO É IMPORTANTE QUE SE COLOQUE DESSA FORMA PARA QUANDO FOR ENCONTRAR A RELAÃO FILOGENÉTICA O PROGRAMA RECONHECER! >Seq1 MAAAGLSSSSCTVFTLSSSFKTRRHLTKTPQNPLLFKRNNSSNRTSHPFLPSRKLCRVQATILREDEEKKVVVEESFQPKTFTHEPVRGSPQSSSPGGLETWAIKLEQSVNVFLTDSVIKILDTLYHDRDYARFFVLETIARVPYFAFISVLHMYESFGWWRRADYLKVHFAESWNEMHHLLIMEELGGNSWWFDRLLAQVIATSYYFMTVLMYALSPRMAYHFSECVESHAFATYDKFIKAQGDDLKKLPAPEVAVKYYTEGDLYLFDEFQTSRAPHSRRPKIENLYDVFLNVRDDEAEHCKTMKACQTHGNLRSPHSYPEDAFEDDTGCDLPQADCEGIVDCIKKSVTSPPSKQNI >Seq2 MAVASTSPLSAKPATAPSPPAPGSGLLALGVRRAPATAAWRRLRVEAIRTQRTEVPVEESAPARDAAAAAPLDGNGAGADGSVVPSSDDSWVVKLEQSFNIFATDSVIMVLKGVYGDRYYARFFALETIARVPYFAFISVLHLYATFGWWRRADYIKVHFAQSWNEFHHLLIMEELGGDSLWFDCFLARFMAFFYYFMTVAMYMLSPRMAYHFSECVERHAYSTYDEFLKLHEEELKRLPAPEAALNYYMNEDLYLFDEFQASRTPGSRRPKIDNLYDVFVNIREDEAEHCKTMKTCQTHGNLRSPHSTPNCLEDDTECVIPENDCEGIVDCVKKSLTK >Seq3 MAVASTSPLSAKPTTATSPPTPAFGFLAPRSRRGRATAWRSLLVEAIRTQREKQGTEGHFEESAAAPLDGVGVGADDPVVPSSDASDWVVKLEQSFNIFATDSVIMVLKGVYRDRYYARFFALETIARVPYFAFISVLHMYSTFGWWRRADYIKVHFAQSWNEFHHLLIMEELGGNSLWIDCFLARFMAFFYYFVTVAMYMLSPRMAYHFSECVERHAYSTYDEFLKLHEEELKRLPAPEAALEYYLNEDLYLFDEFQASRSPGSRRPKIDNLYDVFVNIRDDEAEHCKTMKTCQTHGNLRSPHSTQNCLEADTECVIPENDCEGIVDCVKKSLTK >Seq4 MAISISAMSFGTSVSSYSCFRARSFEKSSVLCNSQNPCRFNSVFPIRKSDGASRCSVSRKSCRVRATLLQENEEEVVVEKSFAPKSFPDNVGGGSNGKPPDDSSSNGLEKWVIKLEQSVNILLTDSVIKILDTLYHNRNYARFFVLETIARVPYFAFISVLHMYESFGWWRRADYMKVHFAESWNEMHHLLIMEELGGNAWWFDRFLAQHIAIFYYFMTVLMYALSPRMAYHFSECVESHAYETYDKFIKDQGEELKNLPAPKIAVDYYTGGDLYLFDEFQTSREPNTRRPKIDNLYDVFMNIRDDEAEHCKTMKACQTHGSLRSPHTDPCDDSEDDTGCSVPQADCIGIVDCIKKSVTDTQVTKR >Seq5 MAMSISPAAHPTSISIRNCKTLSPFPRQNVLRYNPLSPRRSKVSGKLFRVQATVLRENDEKVVVEETFQPKTSTNEERKGGDGEPQDDSSSGTLEKWVIKLEQSINIFLTDSVIKVLDSLYHDRDYARFFVLETIARVPYFAFISVLHMYESFGWWRRADYLKVHFAESWNEMHHLLIMEELGGNAWWFDRFLSQHIAVVYYFMTVLMYAISPRMAYHFSECVESHAFSTYDKFIKAQGEELKKLPAPEVAIKYYTGGDLYLFDEFQTSRAPNT 4) Obtenha um cDNA completo da especie Prunus persica usando dados de ESTs e que apresente a mais alta identidade com a sequencia 5 como isca. Usar o tblastnest (banco de dados) pegar a sequência de aminoácidos do organismo 5 preencher com o nome do organismo Prunus persica APARECERAM 12 RESULTADOS apertar em ALL dessa forma vamos fazer o download no formato fasta para assim levarmos para o CAP3 e apertamos em “contigs” vamos agora alinhar com a sequência 5 (seq. de nucleotídeos) para saber qual desses contigs é mais semelhante a nossa sequência, na parte de cima coloque os dois contigs e na parte de baixo a seq. 5 e marque more dissimilar. O contig 1 é o mais semelhante .Utilize o Expasy para encontrar os códons de iniciação e parada. >Contig1 GACCCTCTTAGGCGGTTGACACCTTCCCTTATACCAACTGACCGAAACCAATTCCAATTTGATTTCCATTTTCTGCTTTTGAATTTTCGTTTCGGTCTCTGCAAAAAACCGTCGGGAATGGCAGCGGCGACTTTGTCTTCCACGGTGTTTGCAATCTCAACCTCCTGCTCTTCTTCGTCTCTCAAAGCAAGGAACTTCAAGAACTTAGCTTGCTCCACGTTTTGTTCTCAGAATCGTATTCCTTACAATCCCATTTCTGCTCGTCCATCAATTTCCAGAATTCGTGCAACTATTCTGCAAGAAGATGAAGAGAAAGTGATAGTGGAGGAATCCTTTGAGTTCAAGGCTTCTTCTCCCTCAGATGAAGTGAAATCAAGCAGTGGGGGCCCATCAGAAAGTTCGTCTTCAAGTACTTTTGAGGGCTGGGTTATTAAATGTGAACAAACCATCAACATCTTCCTTACGGATACAGTCATAAAGATACTTGATACTCTGTACCGTGACCGAGATTATGCAAGGTTTTTTGTACTGGAAACCATTGCAAGGGTTCCTTACTTTGCCTTTATGTCTGTTTTGCACATGTATGAGAGTTTTGGCTGGTGGAGAAGAGCAGATTATCTGAAAGTGCATTTTGCTGAGAGCTGGAATGAGATGCACCACTTGCTTATCATGGAAGAATTGGGGGGAAATGCTTGGTGGTTTGACCGCTTTCTTGCTCAGCATATTGCGATCTTCTATTATTTTATGACGGCCTTTATGTATATAATAAGCCCAAGAATGGCATATCACTTCTCTGAATGTGTAGAGGGCCATGCATTTTCAACTTACGACAAGTTTATCAAGGCCAGAGGAGAGGATTTGAAAAAGTTGCCTCCTCCTGAGGTTGCTGTAAAATACTACACTAGCGGTGACTTGTACTTATTTGATGAATTTCAAACTTCCAGAGCTCCCAATTCTCGAAGGCCAAAAATAGAGAATTTGTACGACGTGTTTCTGAACATAAGAGATGACGAAGCTGAACACTGTAAGACGATGAAGGCTTGCCAGACTCACGGGAACCTGTGGTCTCCTCATTCCCGTGCAGAAGAAGAAGATGACGCCCCGTGCATCATTCCTCAAACAGACTGTGAAGGTATTGTAGATTGTATAACAAAATCCGTGACAACAAAGCCAGAAAATTGATCATATAAGACACAAGAGGGGACGAGGAAATTAGGTAGAAGTAGTGAATGGTATAAGAAATTTGTTTGTATACACAAGAATGTATAGATCCAAAAGAAGCATTTGGTTATCCAATGAAGGTTGCTTTTCTEsse contig apresenta as regiões 5’UTR, 3’ UTRs (em cinza) e o ORF (open reading frame) que começa com o ATG (codon da 1ª metionina – Verde) e termina no TGA (codon de parada – vermelho). Para o cDNA ser considerado completo ainda seria necessário a cauda poli A na extremidade 3’ que não foi encontrada. OBS. Havendo mais de 1 contig deveria ser feito um alinhamento da isca 5 com os contigs para saber qual o mais semelhante. 5) Qual a relação filogenética entre as sequências obtidas através do Clustal? Dada a sequência gênica anotada abaixo, responda as questões de 6 a 10: 6) Trata-se de que gene? De qual organismo? Phaseolus vulgaris – Organismo Actin like - Gene 7) Obtenha a sequência do cDNA (identificando UTRs e ORF) e proteína deduzida. aaggacacgcgtgaagaacaacacgaatgtctcgttgaggcaatgtgctgagaacgtggacaccttcatttaaaaacccaaactcattgaaacttgaggtttgatttgggcgtccaatcccgtgtatcagttttcgccttcagattttgaaagacaatggccgatgctgaagatattcagccactcgtctgcgataatgggacaggaatggttaaggctggatttgccggtgatgatgctccgagggctgtgtttcccagcattgttggtcggccacgtcatactggtgtgatggttgggatggggcaaaaggatgcatatgttggggatgaggctcagtccaagcgtggtatactgactctgaaatatcccattgagcacggtattgtgagcaactgggatgacatggagaagatctggcatcacaccttctacaatgaactccgtgtggccccagaggagcaccctgttctgctcactgaagcccctctcaatccaaaggctaatcgtgagaaaatgacccaaatcatgtttgagaccttcaacacacctgccatgtatgttgccatccaggctgttctgtccctctatgccagcggtcgtacaactggcattgttctggactctggagatggtgtcagccacactgtacccatttatgaaggctatgccctccctcatgccatcctccgtcttgacttagcaggacgtgacctcactgatgccctgatgaaaatcctcactgagcgtggttactccttcactaccacagctgagagagaaattgtaagggatatgaaggaaaaattggcttatattgcccttgattatgaacaagagcttgaaacttcaaaaactagttcatctgttgagaagagctatgagctgcctgatggacaggtcatcaccattggtgccgagcgcttccgttgtcctgaagttctcttccagccatccatgattggaatggaagctgttggcattcatgagacaacatacaattcaattatgaagtgtgatgttgatatcaggaaggatttgtatggaaacatagtcctcagtggtggttctaccatgttccctggtattgcggatagaatgagcaaggaaatcactgccctggccccaagcagcatgaagatcaaggtggttgctccacccgagagaaagtatagtgtctggattggtggctctatcttggcttctctcagcactttccaacagatgtggattgcgaaggcagagtacgatgagtccggtccttcaattgtacacaggaaatgcttttaaatgcagcagaacggcttggtgtcctcgattggcaatgcagtagccccttgcaatgattgaagatctttcttaaaaaatgtttgtgttgttagaacagtgtactaagataaagtggagttttatggcagaattaattctttcatttgttttgttttaacattgttaattgttgaacagttgatcttt ????????????? 8) Quantos exons e introns existem? (Use uma ferramenta que mostre a estrutura gênica) Região transcrita 5 exons interrompidos por 4 introns 9) Qual o perfil de expressão desse gene no experimento de dados transcriptomicos abaixo: Pegue o cDNA 7 Q fazer um blast colar o cDNADatabase: SRA copiar o número do experimento ver o número de sequências e anotar colocar MAX TARGET 20.000 SUBMIT anotar os hits copiar e colar no excel Experiment Library name SRX339933 Leaf Control SRX340118 Leaf Salt SRX340737 Root Control SRX340738 Root Salt O sal inibe a expressão do gene actin tanto em folhas como em raízes com sal, os dados encontrados no gráfico mostram a diferença do número de transcritos. 10) Em relação a informações contidas no promotor desse gene: Que estímulos ambientais poderiam influenciar sua expressão? acatgaaaataaccagcaaatactaatttagcatgttatagtgaagagggcaaaaagttaacaagatattatccttctcgatagaagaaattgacaaagggaatatgcagaaagaaaagagggaggacccaattggatttgcctgaatttcaccacatatctatcttcgaacatgataaaagtgacgtatcttgaggaggaccacacatgcatatgcgactttctaagattctacacatcaatactttatctggtttttccatcatccaacctatagtactgtaagttgacaacgttatatgaagatgctgttgtttttcttagtattccttttcttttgcaccatctgcatttttctcaagcaatttctctgatacttgagatgtttggtggttgagaaaacctccaaagctacctgtccgtagttgaggaaataatgagggttgagaggtaacatgtgatggtgttcgaaaagtaacactgagggatgcatctatttttagttaattcagcaccgtacagtgtgagttaagttgggaaaattattgatgaaaagatgttgcggattatagtgtgctttaattagtatgagagtggtggtgtaggagtagttaaagcatagaaaattaagaaatggaaaagcataatattgtggggtaggaagagggagaagagggccaagtgtatgaaaagtaaagggagcataacacacactctcatggggcataattgcaccccatgctcatgcctaatcattttgcttcccatctcaatcatgtctccatccctaacaatcatgaaccatctttctttccttaatttcctctttttctgcaatttatcactaaaactgttctttatcttttagatagttaagtttaacaacggatgagcccattggggtctgttctccatgttggatcctattgcaccactcctttctccctccagcatcggattctcagctcttgtcgtatataaataaactcccacttcaatatattttgtcctatatagtttgcaaattactacaatctaaatcaatttgaccctactatacgtttctttttaatttctatattcaaatctcatttactaaaatagcataaattatgtatataaactaaaagaagtataataggtcttgtttttttaatcaaaattcctttactatcacaatcacaatgttagctctccagtaataagtcttgtagtgtttgaaaatgtatggacatggctcaatggagttcaattgtctcttgatcatgaatttcatattacgaagtagactttaagcctaagtcaactcctagtcgatgtgggatctttaatacacccctcacgccgagagtgacagttatgactctgataccatatcacgaagtagactttaagtctaactcaaccccataaaatcgactcatggagtgaggttcacaaccacttatatacaatgaaaatttcataatctctaattgatgtgggatctccaacatactttgtaagtgtgatgcggaacacactttataagtgtggtgcaggtaggtcgggagaccggataatcgagtagtaaaatatagttgagcctccaatagtacacaagattgtaagagttttgttttcaagaacatcatattctcacattggttaagttttgaaattgaatgaaaaataagcaaaattggataaaaatataatgtagtgtaggttgaaggaaagcaaaatatttagattaaaaaggataaaaagagtgagaaaagctatggatagaggggatggaagtgcaatgcaagtgagcccagaattttggggcaaacgaatgagcatggaggcaaagaggcaaagaggcagcaggtgcagtgcatgtcgttttgcatggcctgatgattcgtgaaggaatttgttttgtcgtttagtggaaacattatcgtttgttcccatattttgttgaggaaaagacaacacatgtctatgttggtccgtcacagaaagaattgcattgtggtgagaacgtgcagtcttattttctttcttgcatcatttatcatattttggcctataccaacgaacgtaccgttttttgtttttttgcaaaagccatgtttgcatgagacgacccagtgggtggaaagaaggcccaaatggatagtgaagaaatagcggaagtggtaaaaatgggccaagctaataaaagcaatgtgtgtgcggagccacagaaggaaaggacacgcgtgaagaacaacacgaatgtctcgttgaggcaatgtgctgagaacgtggacaccttcatttaaaaacccaaactcattgaaacttgaggtttgatttgggcgtccaatcccgtgtatcagttttcgccttcaggttaatcgctttcttcctcctatctctctaacattctctctaacattctcatgcattgctttttttctctgctttctttttccaattttcaaacaaaatgttcggattctcatataatcacctaccattagtaataatgtcttttctttcgtaactcgatgaatgtgtttaattacttactcttctctcaattgtatccatactaccaggctgtgatcaatttttatttttttgagttaaaattgttccatgttttccccttgtattgtcttgtctttcaatagtttgagaggaaatgactgctttgtgcagattttgaaagacaatggccgatgctgaagatattcagccactcgtctgcgataatgggacaggaatggttaaggtaaaacatgaaagtaggagtgattggatatgtagtatgttcttttaggtgtgggtggtgtctttggatttgtactgatttagtgagtcttttgcactgtactgcgttgcattgcaatttggataatgcaggctggatttgccggtgatgatgctccgagggctgtgtttcccagcattgttggtcggccacgtcatactggtgtgatggttgggatggggcaaaaggatgcatatgttggggatgaggctcagtccaagcgtggtatactgactctgaaatatcccattgagcacggtattgtgagcaactgggatgacatggagaagatctggcatcacaccttctacaatgaactccgtgtggccccagaggagcaccctgttctgctcactgaagcccctctcaatccaaaggctaatcgtgagaaaatgacccaaatcatgtttgagaccttcaacacacctgccatgtatgttgccatccaggctgttctgtccctctatgccagcggtcgtacaactggttagtctttattatgctgtctataaaaaaaacagccaatcctctgcaaggttcacattgcccttgaaaacatggaccagaaaattgaatgtggagaaaagaataacaaaaagggctcaagggtctgtttgattgccttttcatttttcttattttagaaactgttttatattcaacaatcaatttctaaccccagagagggttcattagttttgaaaacgatttcaaaagtaaacaaatggaaacaactttgagatgtgttttcattcttcttgtattctgtgaccctaggtcaagagtgttgttggtcattaaggatggtgctaatgtagtgaggcatgatatgagctgtgttggctttgcagagatttattcgtgatttcaagagctaaacgatttttgaaaatagaaatcaacactccctgattctcaaagttcaaacattttggttttgaccctgctaattttatatgcttaaacagttatctgatatgaagtaattgagttactctaacttggctggttgccgttatggttggtacactaatttgtgtaggcattgttctggactctggagatggtgtcagccacactgtacccatttatgaaggctatgccctccctcatgccatcctccgtcttgacttagcaggacgtgacctcactgatgccctgatgaaaatcctcactgagcgtggttactccttcactaccacagctgagagagaaattgtaagggatatgaaggaaaaattggcttatattgcccttgattatgaacaagagcttgaaacttcaaaaactagttcatctgttgagaagagctatgagctgcctgatggacaggtcatcaccattggtgccgagcgcttccgttgtcctgaagttctcttccagccatccatgattggaatggaagctgttggcattcatgagacaacatacaattcaattatgaagtgtgatgttgatatcaggaaggatttgtatggaaacatagtcctcagtggtggttctaccatgttccctggtattgcggatagaatgagcaaggaaatcactgccctggccccaagcagcatgaagatcaaggtggttgctccacccgagagaaagtatagtgtctggattggtggctctatcttggcttctctcagcactttccaacaggtaaacattctgtgattctaatgaccaaagtaatttgtgttgtgataaacctaaaacggtgatgtctcatgcactttattacaactgcatgacggagacatcaagtgtgtgattgtctttattttagttttagttctgatgctcttatctaaatctcagtttgatagcttggttgcttgaattttgtctccttctcatgacagttgaggttcataggccaattgggatttgtgtaacttactatgcatactaatagcaagcggcatctcatgcatttgggcacacatgctagaatgttttttatatgggtcagcttattaagattattaatgagaatgcaattgtttttggaaatgcagatgtggattgcgaaggcagagtacgatgagtccggtccttcaattgtacacaggaaatgcttttaaatgcagcagaacggcttggtgtcctcgattggcaatgcagtagccccttgcaatgattgaagatctttcttaaaaaatgtttgtgttgttagaacagtgtactaagataaagtggagttttatggcagaattaattctttcatttgttttgttttaacattgttaattgttgaacagttgatctttaatgatatatcaaaaccaaatcagcttattactcattgcttcctcgtaatcatcatattcttctatttaaatcaaagtcagatcctataattctgtatatgtttcaagttaattagaacctcctaatcaaactggtccttggttgggttatatggctgcagagttgagattttaaatacgcattttcactgcaagaattcggagctggttaagatataatgtaagagcatttgtgtttgattttttggttgctgaggcaacattttgaatattatactgaatgtatattactttaaatccttttgctagatgttgaaatttctgaacattttaaatgcttaaaagttggatgtcttacccagcaaacataaacacctacctcggttttagacttactgtcacgttgcctgtgcatctgaatttggttttcagttgaaaacttacatactcagattcaaaaagaaaacaaaaataaagaaacttctcattttagtatttctcatccaaaaaataggttatatatattcaaagaaattagtatgtattatttggtatataaaaatttcaatactttttatcttgtgcactaaatgcctaattaagaaaataaaaagttattgaaataaaataagctaattaaattaaaaacaaaaattaattatttgtgttttttcaatatttaagagaatgaatgcatttcccttaacagatggaacaaaatagattataaaatactattactttataaattattgcacatttaaactataagtgtaagagatacattcaacatataaaaatgagggacgttcagaactgaaactaatttttttttaccgataattgataaaaaaaaaaggtgatgatcaagtaaaatcgaaatttgttggatccatttgaatttttttttagaaccaagcagttctgtttggtttgtggtttaacattttgaaattgatccaaagagaatcgaagagaaatcaattatttaagtattattatagtacttaatatttcactatcatacacaagagaagaagagatgtgtatgcgagagaagaaaattaatatgtctaactagtttcatgtttataaatctgagtaaaagatacgtttaacatatgaataaagtaaaaagaagagtaaaaggaagagttattataatagtgcgaaagttacattattaacaagtgctaaattaaaatcaaagattattgaaaggaaaccatatataatgactaaattttttgatgtttatgttttgaattttgtgaacttatgtgtgcaaagattctgttagagttaataaaatgtacatgctagaagttatttataatttatattacaatgtataaatataatccaatcaccttttttctttttattttatagtaattcatttgttttaaatatttaattatagttaaatgagagaataaaattattcattttgatataacattagagttgttgagaataagaatgaatgacttcttgaataggtcgtgaatgaggtctttgttgttgcgtatattttttgtcttctagttatcgattttgaatagtgttcaatcttctgtcttcgattagtaaagaggagggagtatctgaagaagattatccaatgtttaagtcattactcgatatttgatgattgtagataataaatgttattatatgcatatctttacttgtttggtagtagtggtcaacttatttaaagggataatttgagcttaaaaagtgttgagccaaattaacacgagtagcgttatcaagtctttgtattaatctaacacgaagtctaatgtcactagacttgtcatggatgtagagcattaaatataaaatatgttcctactagggagatgagacctagagaactattgaagtcttcttctccttccttctttcggtcggttaggttgttgggtgatgaacttgaattcttctcgtcctcctgcttctccttcggcactcggtctcgggtgaacaccggatggtgggggtacctgcgaaggcactccgacactcaagttagtaaagcgggtgatcaattttctggtaggtgaacagtaataaatgacgtacctttctacttgggatgtgtgctatttatattattctaatgggcctaccttgttggacccgtttatcggagtggatcttgcttaggattgcattacttaattattaatgatggattagttttactgaccttggtttgagcgttaacggtagtatgggtcggctggctagaccgaccgtgatgtttccacttgtctcggtcgcctcggccgaatctatcgggtctcggccagactccctctgatctcggtctcggacaatcaagtgggtatcggttaggatgaccaagtgtcgatctcgcccatatcgatacaaaatatattcactaataaataaaaatgtatataataaaatcacaaacaaaaactatactaaaaaaatcatttcttttaattttttttttttaatcttcaaatcgaaccaaatcaaacggttaataatttttatattaggttgaaatacattttaacttaaaaaattaaagtacataccgcgaaacatcctaataaaaataattgaacagatataaaacaaagattgtgttaggaatgaatttctaaagtctgaacaaatggactgttaaggttgggattttccccgggttaggtgaataacaagaatattgttcggtctctttcaaaccattctttatttcaaaggatattagcttttaaatgaatataatattatattctattaattctattttataaaattgattaaaaataaaatttacatccaattatataatatagttaagatgttgttacatatagttgatgttaaaaatataaaaaaaaaaatcacagtcgtatacttagtatagagaaaaaatggtagttgaatgaaattaaacat
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