LOCUS Exported 11478 bp ds-DNA circular SYN 29-MAR-2022 DEFINITION encodes S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664) driven by CMV promoter. ACCESSION . VERSION . KEYWORDS pcDNA-dCas9-p300 Core SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11478) AUTHORS Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, Crawford GE, Reddy TE, Gersbach CA TITLE Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. JOURNAL Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 2015 Apr 6. PUBMED 25849900 REFERENCE 2 (bases 1 to 11478) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 29, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11478 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 5..334 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 185..320 /label=SV40 ori /note="SV40 origin of replication" primer_bind 247..266 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 401..1195 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(455..474) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1065..1084 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1369..1490 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1406..1425) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1460..1479 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1539..1555) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1539..1555) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1552..1574) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1563..1579 /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(1587..1617) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1632..1653 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1770..1787) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1941..2529) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2021..2040) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2700..3560) /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 3323..3342 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3561..3665) /gene="bla" /label=AmpR promoter primer_bind complement(3740..3759) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3931..4310 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4311..4514 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4465..4485 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4559..4578 /label=T7 /note="T7 promoter, forward primer" promoter 4559..4577 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4603..4668 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 4675..4695 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4705..8808 /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 8833..8853 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8863..10713 /codon_start=1 /gene="EP300" /product="catalytic histone acetyltransferase core of the human E1A-associated protein p300 (Delvecchio et al., 2013)" /label=p300 core /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD" regulatory 10477..10486 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 10714..10740 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(10765..10782) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 10771..10995 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11041..11469 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11128..11147) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11338..11359 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(11478..20) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" ORIGIN 1 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 61 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 241 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 301 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat 361 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 541 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 901 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 1021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 1081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 1141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 1201 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 1261 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 1321 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 1381 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 1441 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 1501 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 1561 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 1621 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 1681 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 1741 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 1801 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 1861 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 1921 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 1981 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 2041 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 2101 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 2161 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 2221 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 2281 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 2341 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 2401 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 2461 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 2521 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 2581 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 2641 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 2701 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 2761 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 2821 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 2881 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 2941 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 3001 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 3061 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 3121 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 3181 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 3241 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 3301 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 3361 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 3421 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 3481 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 3541 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 3601 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 3661 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc 3721 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 3781 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa 3841 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc 3901 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt 3961 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 4021 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 4081 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 4141 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 4201 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 4261 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt 4321 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 4381 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 4441 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 4501 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta 4561 atacgactca ctatagggag acccaagctg gctagcgcca tggactacaa agaccatgac 4621 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagat ggcccccaag 4681 aagaagagga aggtgggccg cggaatggat aagaaatact caataggctt agctatcggc 4741 acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc 4801 aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta 4861 tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat 4921 acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa 4981 gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga agacaagaag 5041 catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat 5101 ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc 5161 ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga 5221 gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac 5281 aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt 5341 tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag 5401 aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt 5461 aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat 5521 gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca 5581 gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata 5641 actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg 5701 actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt 5761 gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt 5821 tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa 5881 ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat 5941 caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt ttatccattt 6001 ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt 6061 ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca 6121 attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt 6181 gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt 6241 ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa 6301 ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc 6361 ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata 6421 gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt 6481 acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat 6541 gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt 6601 gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa 6661 cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat 6721 aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat 6781 tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca aaaagcacaa 6841 gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct 6901 attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg 6961 cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag 7021 ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga 7081 agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat 7141 ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt 7201 ttaagtgatt atgatgtcga tgccattgtt ccacaaagtt tccttaaaga cgattcaata 7261 gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt 7321 gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc 7381 actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat 7441 aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca 7501 caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag 7561 gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc 7621 tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc 7681 gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat 7741 tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca 7801 accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt 7861 gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt 7921 gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc 7981 aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca 8041 aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt 8101 ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg 8161 aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt 8221 tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa 8281 gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg 8341 atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat 8401 gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac 8461 gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa 8521 atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt 8581 gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta 8641 tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat 8701 cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc 8761 actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgaccc aattgccgga 8821 tccaaggcta gcccgaaaaa gaaacgcaaa gttgggcgcg ccattttcaa accagaagaa 8881 ctacgacagg cactgatgcc aactttggag gcactttacc gtcaggatcc agaatccctt 8941 ccctttcgtc aacctgtgga ccctcagctt ttaggaatcc ctgattactt tgatattgtg 9001 aagagcccca tggatctttc taccattaag aggaagttag acactggaca gtatcaggag 9061 ccctggcagt atgtcgatga tatttggctt atgttcaata atgcctggtt atataaccgg 9121 aaaacatcac gggtatacaa atactgctcc aagctctctg aggtctttga acaagaaatt 9181 gacccagtga tgcaaagcct tggatactgt tgtggcagaa agttggagtt ctctccacag 9241 acactgtgtt gctacggcaa acagttgtgc acaatacctc gtgatgccac ttattacagt 9301 taccagaaca ggtatcattt ctgtgagaag tgtttcaatg agatccaagg ggagagcgtt 9361 tctttggggg atgacccttc ccagcctcaa actacaataa ataaagaaca attttccaag 9421 agaaaaaatg acacactgga tcctgaactg tttgttgaat gtacagagtg cggaagaaag 9481 atgcatcaga tctgtgtcct tcaccatgag atcatctggc ctgctggatt cgtctgtgat 9541 ggctgtttaa agaaaagtgc acgaactagg aaagaaaata agttttctgc taaaaggttg 9601 ccatctacca gacttggcac ctttctagag aatcgtgtga atgactttct gaggcgacag 9661 aatcaccctg agtcaggaga ggtcactgtt agagtagttc atgcttctga caaaaccgtg 9721 gaagtaaaac caggcatgaa agcaaggttt gtggacagtg gagagatggc agaatccttt 9781 ccataccgaa ccaaagccct ctttgccttt gaagaaattg atggtgttga cctgtgcttc 9841 tttggcatgc atgttcaaga gtatggctct gactgccctc cacccaacca gaggagagta 9901 tacatatctt acctcgatag tgttcatttc ttccgtccta aatgcttgag gactgcagtc 9961 tatcatgaaa tcctaattgg atatttagaa tatgtcaaga aattaggtta cacaacaggg 10021 catatttggg catgtccacc aagtgaggga gatgattata tcttccattg ccatcctcct 10081 gaccagaaga tacccaagcc caagcgactg caggaatggt acaaaaaaat gcttgacaag 10141 gctgtatcag agcgtattgt ccatgactac aaggatattt ttaaacaagc tactgaagat 10201 agattaacaa gtgcaaagga attgccttat ttcgagggtg atttctggcc caatgttctg 10261 gaagaaagca ttaaggaact ggaacaggag gaagaagaga gaaaacgaga ggaaaacacc 10321 agcaatgaaa gcacagatgt gaccaaggga gacagcaaaa atgctaaaaa gaagaataat 10381 aagaaaacca gcaaaaataa gagcagcctg agtaggggca acaagaagaa acccgggatg 10441 cccaatgtat ctaacgacct ctcacagaaa ctatatgcca ccatggagaa gcataaagag 10501 gtcttctttg tgatccgcct cattgctggc cctgctgcca actccctgcc tcccattgtt 10561 gatcctgatc ctctcatccc ctgcgatctg atggatggtc gggatgcgtt tctcacgctg 10621 gcaagggaca agcacctgga gttctcttca ctccgaagag cccagtggtc caccatgtgc 10681 atgctggtgg agctgcacac gcagagccag gactacccgt acgacgttcc ggactacgct 10741 tcttgagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg 10801 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct 10861 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg 10921 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg 10981 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc 11041 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 11101 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 11161 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 11221 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 11281 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 11341 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 11401 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 11461 acgcgaatta attctgtg //