LOCUS Exported 14933 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expression of AB1-fused-dcas9 plasmids. ACCESSION . VERSION . KEYWORDS pHR-PGK-ABI-dCas9-P2A-mcherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14933) AUTHORS Wang H, Xu X, Nguyen CM, Liu Y, Gao Y, Lin X, Daley T, Kipniss NH, La Russa M, Qi LS TITLE CRISPR-Mediated Programmable 3D Genome Positioning and Nuclear Organization. JOURNAL Cell. 2018 Nov 15;175(5):1405-1417.e14. doi: 10.1016/j.cell.2018.09.013. Epub 2018 Oct 11. PUBMED 30318144 REFERENCE 2 (bases 1 to 14933) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14933 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 95..112 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(182..202) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 187..516 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 367..502 /label=SV40 ori /note="SV40 origin of replication" primer_bind 429..448 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 1650..1715 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 1845..1865 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 2290..2424 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2327..2346) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2381..2400 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 2592..3225 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 3272..3397 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3894..4127 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 4312..4356 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 4641..4758 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 4823..5322 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4858..4881) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5255..5274 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6313..10416 /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 10420..10446 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 10465..10485 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10492..10512 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10561..10617 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 10618..11328 /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(10763..10781) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 11031..11050 /label=mCherry-F /note="mCherry, forward primer" misc_feature 11388..11976 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(11441..11461) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(11859..11870) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(11979..11995) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11980..11996) /label=pBluescriptKS /note="For pBluescript vector" LTR 12086..12319 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(12344..12362) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(12345..12362) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(12652..12671) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(12885..12904) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 13004..13026 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(13064..13082) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 13150..13254 /gene="bla" /label=AmpR promoter CDS 13255..14115 /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(13473..13492) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 14286..14874 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 14775..14794 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 181 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 241 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 361 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 421 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 481 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttggacacaa gacaggcttg 541 cgagatatgt ttgagaatac cactttatcc cgcgtcaggg agaggcagtg cgtaaaaaga 601 cgcggactca tgtgaaatac tggtttttag tgcgccagat ctctataatc tcgcgcaacc 661 tattttcccc tcgaacactt tttaagccgt agataaacag gctgggacac ttcacatgag 721 cgaaaaatac atcgtcacct gggacatgtt gcagatccat gcacgtaaac tcgcaagccg 781 actgatgcct tctgaacaat ggaaaggcat tattgccgta agccgtggcg gtctgtaccg 841 ggtgcgttac tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg tatttccagc 901 tacgatcacg acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg cgatggcgaa 961 ggcttcatcg ttattgatga cctggtggat accggtggta ctgcggttgc gattcgtgaa 1021 atgtatccaa aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg tccgctggtt 1081 gatgactatg ttgttgatat cccgcaagat acctggattg aacagccgtg ggatatgggc 1141 gtcgtattcg tcccgccaat ctccggtcgc taatcttttc aacgcctggc actgccgggc 1201 gttgttcttt ttaacttcag gcgggttaca atagtttcca gtaagtattc tggaggctgc 1261 atccatgaca caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta aacatcctga 1321 aacctcgacg ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc agtcggccct 1381 tgatggtaaa accatccctc actggtatcg catgattaac cgtctgatgt ggatctggcg 1441 cggcattgac ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga gcgatgccga 1501 acgtaccgac gatgatttat acgatacggt gattggctac cgtggcggca actggattta 1561 tgagtgggcc ccggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca 1621 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg 1681 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat 1741 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat 1801 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa 1861 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt 1921 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc 1981 tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat aacagttata 2041 atcataacat actgtttttt cttactccac acaggcatag agtgtctgct attaataact 2101 atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt 2161 tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt agaggtttta 2221 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 2281 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 2341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 2401 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa 2461 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc 2521 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca 2581 aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg 2641 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 2701 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 2761 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 2821 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 2881 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 2941 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 3001 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 3061 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 3121 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 3181 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag 3241 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga 3301 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag 3361 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag 3421 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca 3481 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac 3541 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga 3601 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga 3661 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac 3721 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac 3781 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca 3841 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct 3901 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg 3961 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg 4021 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag 4081 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt 4141 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa 4201 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat 4261 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa 4321 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat 4381 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata 4441 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt 4501 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt 4561 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgccaa 4621 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg 4681 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt 4741 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggatcgat 4801 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca 4861 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca 4921 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca 4981 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg 5041 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag 5101 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc 5161 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc 5221 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg 5281 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta 5341 cgcgtctgca ggatatcaag cttgcggtac cgcgggcccg gggccaccat gacgcgtgtg 5401 cctttgtatg gttttacttc gatttgtgga agaagacctg agatggaagc tgctgtttcg 5461 actataccaa gattccttca atcttcctct ggttcgatgt tagatggtcg gtttgatcct 5521 caatccgccg ctcatttctt cggtgtttac gacggccatg gcggttctca ggtagcgaac 5581 tattgtagag agaggatgca tttggctttg gcggaggaga tagctaagga gaaaccgatg 5641 ctctgcgatg gtgatacgtg gctggagaag tggaagaaag ctcttttcaa ctcgttcctg 5701 agagttgact cggagattga gtcagttgcg ccggagacgg ttgggtcaac gtcggtggtt 5761 gccgttgttt tcccgtctca catcttcgtc gctaactgcg gtgactctag agccgttctt 5821 tgccgcggca aaactgcact tccattatcc gttgaccata aaccggatag agaagatgaa 5881 gctgcgagga ttgaagccgc aggagggaaa gtgattcagt ggaatggagc tcgtgttttc 5941 ggtgttctcg ccatgtcgag atccattggc gatagatact tgaaaccatc catcattcct 6001 gatccggaag tgacggctgt gaagagagta aaagaagatg attgtctgat tttggcgagt 6061 gacggggttt gggatgtaat gacggatgaa gaagcgtgtg agatggcaag gaagcggatt 6121 ctcttgtggc acaagaaaaa cgcggtggct ggggatgcat cgttgctcgc ggatgagcgg 6181 agaaaggaag ggaaagatcc tgcggcgatg tccgcggctg agtatttgtc aaagctggcg 6241 atacagagag gaagcaaaga caacataagt gtggtggtgg ttgatttgaa gcatatgggt 6301 ggaggctccg ggatggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg 6361 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc 6421 aacaccgacc ggcacagcat caagaagaac ctgatcggcg ccctgctgtt cgacagcgga 6481 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag 6541 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc 6601 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac 6661 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac 6721 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg 6781 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc 6841 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc 6901 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg 6961 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc 7021 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc 7081 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac 7141 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg 7201 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc 7261 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa 7321 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag 7381 aacggctacg ccggctacat cgatggcgga gccagccagg aagagttcta caagttcatc 7441 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag 7501 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg 7561 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac 7621 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc 7681 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg 7741 aacttcgagg aagtggtgga caagggcgcc agcgcccaga gcttcatcga gcggatgacc 7801 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag 7861 tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag 7921 cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg acctgctgtt caagaccaac 7981 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac 8041 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat 8101 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg 8161 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg 8221 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac 8281 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc 8341 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg 8401 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag 8461 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc 8521 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc 8581 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac 8641 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg 8701 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg 8761 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac 8821 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga taacaaagtg 8881 ctgactcgga gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg 8941 aagaagatga agaactactg gcgccagctg ctgaatgcca agctgattac ccagaggaag 9001 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc 9061 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac 9121 tcccggatga acactaagta cgacgagaac gacaaactga tccgggaagt gaaagtgatc 9181 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc 9241 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc 9301 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac 9361 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac 9421 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag 9481 atccggaagc ggcctctgat cgagacaaac ggcgaaacag gcgagatcgt gtgggataag 9541 ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc cccaagtgaa tatcgtgaaa 9601 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc 9661 gacaagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc 9721 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa 9781 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag 9841 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc 9901 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct 9961 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg 10021 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag 10081 ctgtttgtgg aacagcacaa acactacctg gacgagatca tcgagcagat cagcgagttc 10141 tccaagagag tgatcctggc cgacgctaat ctggacaagg tgctgagcgc ctacaacaag 10201 cacagagaca agcctatcag agagcaggcc gagaatatca tccacctgtt taccctgacc 10261 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac 10321 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac 10381 gagacacgga tcgacctgtc tcagctggga ggcgacgcct atccctatga cgtgcccgat 10441 tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca aggtggaaga tcctaagaaa 10501 aagcggaaag tggacggcat tggtagtggg agcaacggca gcagcggatc cggaagcgga 10561 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg 10621 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg 10681 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc 10741 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc 10801 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc 10861 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg 10921 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc 10981 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg 11041 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc 11101 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag 11161 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac 11221 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc 11281 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgact 11341 ctagagtcga cctgcaggca tgcaagcttg atatcaagct tatcgataat caacctctgg 11401 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 11461 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 11521 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 11581 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 11641 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 11701 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 11761 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca 11821 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 11881 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 11941 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgaggga 12001 attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 12061 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat 12121 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 12181 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 12241 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 12301 gtgtggaaaa tctctagcag catctagaat taattccgtg tattctatag tgtcacctaa 12361 atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct aacgacaata 12421 tgtacaagcc taattgtgta gcatctggct tactgaagca gaccctatca tctctctcgt 12481 aaactgccgt cagagtcggt ttggttggac gaaccttctg agtttctggt aacgccgtcc 12541 cgcacccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag atcctctacg 12601 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg 12661 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg 12721 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg 12781 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa 12841 tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc tcagtacaat ctgctctgat 12901 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 12961 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 13021 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta 13081 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 13141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 13201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 13261 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 13321 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 13381 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 13441 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 13501 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 13561 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 13621 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 13681 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 13741 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 13801 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 13861 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 13921 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 13981 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 14041 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 14101 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 14161 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 14221 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 14281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 14341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 14401 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 14461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 14521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 14581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 14641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 14701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 14761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 14821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 14881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca cat //