LOCUS Exported 10742 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions of interest.. ACCESSION . VERSION . KEYWORDS 3xFLAG-dCas9/MSCV-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10742) AUTHORS Fujita T, Kitaura F, Oji A, Tanigawa N, Yuno M, Ikawa M, Taniuchi I, Fujii H TITLE Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis. JOURNAL Genes Cells. 2018 Feb 26. doi: 10.1111/gtc.12573. PUBMED 29480524 REFERENCE 2 (bases 1 to 10742) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10742 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1462..1527 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1531..5634 /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5647..5667 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter 5839..6338 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5874..5897) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6271..6290 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 6356..6365 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6362..7081 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 6362..7078 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6407..6428) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6668..6687) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7015..7036 /label=EGFP-C /note="EGFP, forward primer" LTR 7135..7649 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 7811..7827 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7835..7865) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7880..7901 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8018..8035) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8189..8777) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8269..8288) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8948..9808) /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 9571..9590 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9809..9913) /gene="bla" /label=AmpR promoter primer_bind 9981..9999 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(10037..10059) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 10159..10178 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 10372..10394 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 10522..10541 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 10741..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ggtttaagga tccgtttagt 1441 gaaccgtcag aattaaccat ggactacaaa gaccatgacg gtgattataa agatcatgac 1501 atcgattaca aggatgacga tgacaagctc atggacaaga agtactccat tgggctcgct 1561 atcggcacaa acagcgtcgg ctgggccgtc attacggacg agtacaaggt gccgagcaaa 1621 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc 1681 ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc 1741 agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg 1801 gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat 1861 aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa 1921 aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac 1981 ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc 2041 gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag 2101 acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca 2161 atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct 2221 ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc 2281 aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc 2341 tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt 2401 ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg 2461 gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa 2521 gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt 2581 ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag 2641 gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg 2701 gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc 2761 ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac 2821 ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac 2881 tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa 2941 gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc 3001 ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa 3061 cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc 3121 acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac 3181 ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa 3241 aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc 3301 ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag 3361 gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag 3421 atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag 3481 ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc 3541 cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac 3601 aggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa 3661 gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc 3721 ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta 3781 atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc 3841 cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa 3901 ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag 3961 ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc 4021 aatcggctct ccgactacga cgtggatgcc atcgtgcccc agtcttttct caaagatgat 4081 tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc 4141 ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa 4201 ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag 4261 ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac 4321 gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt 4381 cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt 4441 cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat 4501 gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac 4561 ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc 4621 aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt 4681 acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga 4741 gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg 4801 caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc 4861 ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa 4921 tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag 4981 aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag 5041 cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc 5101 aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg 5161 aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct 5221 aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa 5281 gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc 5341 gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg 5401 ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc 5461 cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc 5521 atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag 5581 tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg 5641 gctgacccca agaagaagag gaaggtgtga aagggttcga tccctaccgg atcgaattcc 5701 agctgagcgc cggtcgctac cattaccagt tggtctggtg tcaaaaataa taataaccgg 5761 gcaggccatg tctgcccgta tttcgcgtaa ggaaatccat tatgtactat ttaaactcga 5821 gcggccacga attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg 5881 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt 5941 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt 6001 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt 6061 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat 6121 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg 6181 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg 6241 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg 6301 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttgccac 6361 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 6421 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 6481 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 6541 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 6601 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 6661 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 6721 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 6781 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 6841 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 6901 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 6961 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 7021 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta 7081 aagcggccat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa 7141 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa 7201 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata 7261 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga 7321 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg 7381 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc 7441 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac 7501 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc 7561 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga 7621 ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac 7681 attctgaggg taggagtcga atattaagta atcctgactc aattagccac tgttttgaat 7741 ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag 7801 aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 7861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 7921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 7981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 8041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 8101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 8161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 8221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 8281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 8341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 8401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 8461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 8521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 8581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 8641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 8701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 8761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 8821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 8881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 8941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 9001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 9061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 9121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 9181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 9241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 9301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 9361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 9421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 9481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 9541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 9601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 9661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 9721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 9781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 9841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 9901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta 9961 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg 10021 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt 10081 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc 10141 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt 10201 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca 10261 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg 10321 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac 10381 gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc 10441 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg 10501 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt 10561 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa 10621 agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc 10681 tatagagtac gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg 10741 aa //