LOCUS Exported 10155 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of dCas9-DNMT3A fusion with T2A-PuroR and specific sgRNA for targeted DNA methylation of BACH2 promoter in human cells; for use as a control. ACCESSION . VERSION . KEYWORDS pdCas9-DNMT3A-PuroR_BACH2-sgRNA8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10155) AUTHORS Vojta A, Dobrinic P, Tadic V, Bockor L, Korac P, Julg B, Klasic M, Zoldos V TITLE Repurposing the CRISPR-Cas9 system for targeted DNA methylation. JOURNAL Nucleic Acids Res. 2016 Mar 11. pii: gkw159. PUBMED 26969735 REFERENCE 2 (bases 1 to 10155) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10155 /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 271..346 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 443..728 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 730..1007 /label=chicken beta-actin promoter intron 1008..1236 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" regulatory 1248..1257 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1257..1322 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1329..1349 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1374..5474 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5475..5522 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 5550..6482 /codon_start=1 /product="catalytic domain of DNA methyltransferase 3A" /label=DNMT3A catalytic domain /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR LARQRLLGRSWSVPVIRHLFAPLKEYFACV" CDS 6510..6563 /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 6564..7163 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6564..6583) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind complement(7196..7213) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7202..7409 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 7418..7558 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 7418..7547 /label=AAV2 ITR rep_origin 7633..8088 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7720..7739) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7930..7951 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(8105..8124) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8224..8246 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8284..8302) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8370..8474 /gene="bla" /label=AmpR promoter CDS 8475..9335 /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(8693..8712) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9506..10094 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9995..10014 /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 aatgtagcga ttgagagtgt gttttagagc tagaaatagc aagttaaaat 301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga 361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca 421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac 481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac 541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc 661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc 781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg 841 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc 901 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga 961 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga 1021 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga 1081 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat 1141 tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat 1201 tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc accatggact 1261 ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac gatgacgata 1321 agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga 1381 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg 1441 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca 1501 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc 1561 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag 1621 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt 1681 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg 1741 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg 1801 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt 1861 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc 1921 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca 1981 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa 2041 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc 2101 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac 2161 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg 2221 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg 2281 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga 2341 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc 2401 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg 2461 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg 2521 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga 2581 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc 2641 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc 2701 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct 2761 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca 2821 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca 2881 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc 2941 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc 3001 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc 3061 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg 3121 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca 3181 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga 3241 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg 3301 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc 3361 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga 3421 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct 3481 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca 3541 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg 3601 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg 3661 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga 3721 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca 3781 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg 3841 tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct atcgtgcctc 3901 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc 3961 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc 4021 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg 4081 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa 4141 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg 4201 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt 4261 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg 4321 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc 4381 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca 4441 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga 4501 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg 4561 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc 4621 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg 4681 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga 4741 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc 4801 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc 4861 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag 4921 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt 4981 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa 5041 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga 5101 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc 5161 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg 5221 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag 5281 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct 5341 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg 5401 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc 5461 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggacaggcc aaaaagaaaa 5521 agctcgaggg cggaggcggg agcggatccc cctcccggct ccagatgttc ttcgctaata 5581 accacgacca ggaatttgac cctccaaagg tttacccacc tgtcccagct gagaagagga 5641 agcccatccg ggtgctgtct ctctttgatg gaatcgctac agggctcctg gtgctgaagg 5701 acttgggcat tcaggtggac cgctacattg cctcggaggt gtgtgaggac tccatcacgg 5761 tgggcatggt gcggcaccag gggaagatca tgtacgtcgg ggacgtccgc agcgtcacac 5821 agaagcatat ccaggagtgg ggcccattcg atctggtgat tgggggcagt ccctgcaatg 5881 acctctccat cgtcaaccct gctcgcaagg gcctctacga gggcactggc cggctcttct 5941 ttgagttcta ccgcctcctg catgatgcgc ggcccaagga gggagatgat cgccccttct 6001 tctggctctt tgagaatgtg gtggccatgg gcgttagtga caagagggac atctcgcgat 6061 ttctcgagtc caaccctgtg atgattgatg ccaaagaagt gtcagctgca cacagggccc 6121 gctacttctg gggtaacctt cccggtatga acaggccgtt ggcatccact gtgaatgata 6181 agctggagct gcaggagtgt ctggagcatg gcaggatagc caagttcagc aaagtgagga 6241 ccattactac gaggtcaaac tccataaagc agggcaaaga ccagcatttt cctgtgttca 6301 tgaatgagaa agaggacatc ttatggtgca ctgaaatgga aagggtattt ggtttcccag 6361 tccactatac tgacgtctcc aacatgagcc gcttggcgag gcagagactg ctgggccggt 6421 catggagcgt gccagtcatc cgccacctct tcgctccgct gaaggagtat tttgcgtgtg 6481 tgtccggccg gcccgaattc ggcagtggag agggcagagg aagtctgcta acatgcggtg 6541 acgtcgagga gaatcctggc ccaatgaccg agtacaagcc cacggtgcgc ctcgccaccc 6601 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca 6661 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct 6721 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg 6781 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc 6841 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc 6901 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg 6961 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc 7021 gcgccggggt gcccgccttc ctggagacct ccgcgcccca caacctcccc ttctacgagc 7081 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca 7141 tgacccgcaa gcccggtgcc tgagaattca attctaacta gagctcgctg atcagcctcg 7201 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 7261 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 7321 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 7381 tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg 7441 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 7501 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg 7561 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa 7621 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 7681 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 7741 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 7801 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt 7861 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 7921 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt 7981 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 8041 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc 8101 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg 8161 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 8221 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa 8281 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 8341 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 8401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 8461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 8521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 8581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 8641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 8701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 8761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 8821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 8881 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 8941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 9001 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 9061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 9121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 9181 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 9241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 9301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 9361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 9421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 9481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 9541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 9601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 9661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 9721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 9781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 9841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 9901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 9961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 10021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 10081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 10141 cttttgctca catgt //