LOCUS Exported 9474 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION t1 gRNA (ATGAGAATCAAGGCGGTCGA). ACCESSION . VERSION . KEYWORDS pCas9–mKate2ps–T1gRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9474) AUTHORS Moore R, Spinhirne A, Lai MJ, Preisser S, Li Y, Kang T, Bleris L TITLE CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells. JOURNAL Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18. PUBMED 25527740 REFERENCE 2 (bases 1 to 9474) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9474 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 600..609 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 606..4709 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4722..4742 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4746..4799 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 4812..5507 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" CDS 5526..5645 /codon_start=1 /product="PEST degradation sequence from mouse ornithine decarboxylase" /label=hPEST /note="human codon-optimized" /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV" polyA_signal 5779..5900 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5816..5835) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5870..5889 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5907..6362) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6044..6065) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6256..6275 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6389..6493 /gene="bla" /label=AmpR promoter promoter 6495..6852 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6518..6538) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6703..6838 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6765..6784 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6887..7681 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6941..6960) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7551..7570 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7869..7888) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7913..7960 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" promoter 8053..8293 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 8053..8073 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 8224..8243 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 8322..8397 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" protein_bind complement(8409..8433) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" rep_origin 8772..9360 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9261..9280 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccttg 601 ccaccatgga caagaagtac tccattgggc tcgatatcgg cacaaacagc gtcggctggg 661 ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg 721 atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc ggggagacgg 781 ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga 841 tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc 901 ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct 961 ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga 1021 ggaagaagct tgtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg 1081 cgcatatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca 1141 gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt ttcgaagaga 1201 acccgatcaa cgcatccgga gttgacgcca aagcaatcct gagcgctagg ctgtccaaat 1261 cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg 1321 gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg 1381 ccgaagatgc caagcttcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc 1441 tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg 1501 ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg 1561 ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg 1621 tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct 1681 acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca 1741 tcttggaaaa aatggacggc accgaggagc tgctggtaaa gcttaacaga gaagatctgt 1801 tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac 1861 tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa 1921 agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc gcccggggaa 1981 attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc tggaacttcg 2041 aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg 2101 ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca 2161 cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat 2221 tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag 2281 ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc gactctgttg 2341 aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga 2401 aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca 2461 ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt 2521 acgctcatct cttcgacgac aaagtcatga aacagctcaa gaggcgccga tatacaggat 2581 gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt ggaaagacaa 2641 tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg 2701 atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca 2761 gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc 2821 agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata 2881 tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg 2941 aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac 3001 acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg 3061 gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac tacgacgtgg 3121 atcatatcgt gccccagtct tttctcaaag atgattctat tgataataaa gtgttgacaa 3181 gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa 3241 tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata 3301 atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa 3361 ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca 3421 tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga 3481 agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg agagagatca 3541 acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca 3601 aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg tacgatgtta 3661 ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt 3721 acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga 3781 agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg 3841 atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg 3901 aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc 3961 tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag 4021 tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa 4081 gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa aaaaacccca 4141 tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc attaagcttc 4201 ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg 4261 agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg 4321 ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag cagctgttcg 4381 tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa ttctccaaaa 4441 gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg 4501 ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg 4561 gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta 4621 caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa 4681 gaatcgacct ctctcagctc ggtggagaca gcagggctga ccccaagaag aagaggaagg 4741 tggctgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggccctt 4801 ccggagggcc catggtgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg 4861 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg 4921 agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg 4981 acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca 5041 tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga gtcaccacat 5101 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca 5161 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga 5221 aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc ggcctggaag 5281 gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttgaaga 5341 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg 5401 acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg 5461 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagaggt ggaggaggtt 5521 ccggatctca cggcttccct cccgaggtgg aggagcaggc cgccggcacc ctgcccatga 5581 gctgcgccca ggagagcggc atggatagac accctgctgc ttgcgccagc gccaggatca 5641 acgtctaagg ccggccagcg gccgcgactc tagatcataa tcagccatac cacatttgta 5701 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 5761 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 5821 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 5881 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc 5941 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 6001 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 6061 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 6121 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 6181 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 6241 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 6301 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 6361 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 6421 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 6481 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 6541 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 6601 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 6661 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 6721 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6781 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6841 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 6901 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 6961 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 7021 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 7081 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 7141 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 7201 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 7261 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 7321 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 7381 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 7441 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 7501 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 7561 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 7621 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 7681 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 7741 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 7801 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 7861 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 7921 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 7981 cagggctggc actctgtcga taccccaccg agacctcagt cgactggatc cggtaccaag 8041 gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat acgatacaag 8101 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat 8161 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa 8221 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc 8281 ttgtggaaag gacgaaacac cgtgagaatc aaggcggtcg agttttagag ctagaaatag 8341 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt 8401 ttttctagac ccagctttct tgtacaaagt tggcattaaa gggcgaattc cagcacactg 8461 gcggccgtta ctagtggatc cgccacccca ccgagacccc attggggcca atacgcccgc 8521 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 8581 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 8641 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 8701 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 8761 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 8821 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 8881 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 8941 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 9001 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 9061 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 9121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 9181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 9241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 9301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 9361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 9421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //