LOCUS Exported 10003 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cas9 from S.pyogenes with CMV-mcherry cassette, and cloning backbone for sgRNA. ACCESSION . VERSION . KEYWORDS px330-mcherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10003) AUTHORS Wu Y, Liang D, Wang Y, Bai M, Tang W, Bao S, Yan Z, Li D, Li J TITLE Correction of a genetic disease in mouse via use of CRISPR-Cas9. JOURNAL Cell Stem Cell. 2013 Dec 5;13(6):659-62. doi: 10.1016/j.stem.2013.10.016. PUBMED 24315440 REFERENCE 2 (bases 1 to 10003) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10003 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 268..343 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 440..725 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 727..1004 /label=chicken beta-actin promoter intron 1005..1233 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" regulatory 1245..1254 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1254..1319 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1326..1346 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1371..5471 /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5472..5519 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" primer_bind complement(5547..5564) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5553..5760 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 5771..5852 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 5796..5815 /label=SV40pA-R /note="SV40 polyA, reverse primer" misc_feature 5976..6041 /label=MCS /note="multiple cloning site" CDS complement(6042..6749) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(6317..6336) /label=mCherry-F /note="mCherry, forward primer" primer_bind 6586..6604 /label=mCherry-R /note="mCherry, reverse primer" regulatory 6746..6755 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(6794..6997) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(6823..6843) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 6998..7301 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin 7481..7936 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7568..7587) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7778..7799 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7953..7972) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8072..8094 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8132..8150) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8218..8322 /gene="bla" /label=AmpR promoter CDS 8323..9183 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8541..8560) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9354..9942 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9843..9862 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac 421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt 661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc 1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg 1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag 1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac 1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata 1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga 1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt 1381 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt 1441 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga 1501 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga 1561 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga 1621 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct 1681 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg 1741 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca 1801 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc 1861 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt 1921 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg 1981 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc 2041 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga 2101 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc 2161 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc 2221 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca 2281 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat 2341 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg 2401 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg 2461 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg 2521 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct 2581 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc 2641 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga 2701 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga 2761 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg 2821 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg 2881 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga 2941 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga 3001 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga 3061 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag 3121 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg 3181 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac 3241 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg 3301 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga 3361 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt 3421 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta 3481 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg 3541 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg 3601 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca 3661 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg 3721 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc 3781 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg 3841 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga 3901 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg 3961 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc 4021 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga 4081 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc 4141 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg 4201 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg 4261 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc 4321 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg 4381 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga 4441 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact 4501 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga 4561 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga 4621 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct 4681 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg 4741 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg 4801 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg 4861 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca 4921 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg 4981 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg 5041 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc 5101 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact 5161 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg 5221 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc 5281 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca 5341 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg 5401 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc 5461 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt 5521 aagaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 5581 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 5641 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 5701 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga 5761 gcggccgcat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat 5821 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt 5881 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg 5941 gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc ggtaccgtcg 6001 actgcagaat tcgaagcttg agctcgagat ctgagtccgg acttgtacag ctcgtccatg 6061 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg 6121 ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc 6181 ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc 6241 agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag 6301 gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg 6361 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc 6421 accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac 6481 agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg 6541 aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc 6601 ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg 6661 tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc 6721 atgttatcct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cggatctgac 6781 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt 6841 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa 6901 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct 6961 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata 7021 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg 7081 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat 7141 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc 7201 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg 7261 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc 7321 tatgaactaa tgaccccgta attgattact atgcggccgc aaatgggcgg cctcagtgag 7381 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct 7441 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca 7501 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 7561 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 7621 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 7681 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt 7741 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 7801 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg 7861 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 7921 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 7981 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 8041 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 8101 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 8161 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 8221 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 8281 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 8341 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 8401 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 8461 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 8521 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 8581 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 8641 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 8701 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 8761 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 8821 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 8881 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 8941 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 9001 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca 9061 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 9121 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 9181 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 9241 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 9301 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 9361 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 9421 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 9481 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 9541 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 9601 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 9661 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 9721 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 9781 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 9841 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 9901 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 9961 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt //