LOCUS Exported 14914 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION dCas9 fused to 24 copies of the GCN4 peptide v4, which is part of the SunTag system. ACCESSION . VERSION . KEYWORDS pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14914) AUTHORS Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD TITLE A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. JOURNAL Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039. PUBMED 25307933 REFERENCE 2 (bases 1 to 14914) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14914 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(85..105) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 90..419 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 270..405 /label=SV40 ori /note="SV40 origin of replication" intron 1553..1618 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 1748..1768 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 2193..2327 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2230..2249) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2284..2303 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 2495..3128 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 3175..3300 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3797..4030 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 4215..4259 /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 4544..4661 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 4724..4934 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4793..4928 /label=SV40 ori /note="SV40 origin of replication" CDS 4950..4970 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 5019..5028 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5025..9128 /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 9132..9158 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 9177..9197 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9204..9224 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9303..9359 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9375..9431 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9447..9503 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9519..9575 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9591..9647 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9663..9719 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9735..9791 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9807..9863 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9879..9935 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 9951..10007 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10023..10079 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10095..10151 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10167..10223 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10239..10295 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10311..10367 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10383..10439 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10455..10511 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10527..10583 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10599..10655 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10671..10727 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10743..10799 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10815..10871 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 10959..11015 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 11055..11075 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 11085..11141 /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 11154..11855 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" LTR 11970..12203 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(12228..12246) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(12229..12246) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(12536..12555) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(12769..12788) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 12888..12910 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(12948..12966) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 13034..13138 /gene="bla" /label=AmpR promoter CDS 13139..13999 /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(13357..13376) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 14170..14758 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 14659..14678 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 14912..15 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 61 tggccgattc attaatgcag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 421 agcttggaca caagacaggc ttgcgagata tgtttgagaa taccacttta tcccgcgtca 481 gggagaggca gtgcgtaaaa agacgcggac tcatgtgaaa tactggtttt tagtgcgcca 541 gatctctata atctcgcgca acctattttc ccctcgaaca ctttttaagc cgtagataaa 601 caggctggga cacttcacat gagcgaaaaa tacatcgtca cctgggacat gttgcagatc 661 catgcacgta aactcgcaag ccgactgatg ccttctgaac aatggaaagg cattattgcc 721 gtaagccgtg gcggtctgta ccgggtgcgt tactggcgcg tgaactgggt attcgtcatg 781 tcgataccgt ttgtatttcc agctacgatc acgacaacca gcgcgagctt aaagtgctga 841 aacgcgcaga aggcgatggc gaaggcttca tcgttattga tgacctggtg gataccggtg 901 gtactgcggt tgcgattcgt gaaatgtatc caaaagcgca ctttgtcacc atcttcgcaa 961 aaccggctgg tcgtccgctg gttgatgact atgttgttga tatcccgcaa gatacctgga 1021 ttgaacagcc gtgggatatg ggcgtcgtat tcgtcccgcc aatctccggt cgctaatctt 1081 ttcaacgcct ggcactgccg ggcgttgttc tttttaactt caggcgggtt acaatagttt 1141 ccagtaagta ttctggaggc tgcatccatg acacaggcaa acctgagcga aaccctgttc 1201 aaaccccgct ttaaacatcc tgaaacctcg acgctagtcc gccgctttaa tcacggcgca 1261 caaccgcctg tgcagtcggc ccttgatggt aaaaccatcc ctcactggta tcgcatgatt 1321 aaccgtctga tgtggatctg gcgcggcatt gacccacgcg aaatcctcga cgtccaggca 1381 cgtattgtga tgagcgatgc cgaacgtacc gacgatgatt tatacgatac ggtgattggc 1441 taccgtggcg gcaactggat ttatgagtgg gccccggatc tttgtgaagg aaccttactt 1501 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat 1561 aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat 1621 tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct 1681 gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc 1741 tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct 1801 aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac 1861 cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt 1921 tataagtagg cataacagtt ataatcataa catactgttt tttcttactc cacacaggca 1981 tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg 2041 taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc 2101 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc 2161 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt 2221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 2281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc 2341 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc 2401 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt 2461 gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca ctcccaaaga 2521 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag 2581 aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta 2641 gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta 2701 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt 2761 gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac 2821 tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc 2881 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc 2941 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg 3001 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg 3061 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat 3121 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct 3181 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg 3241 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc 3301 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga 3361 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca 3421 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa 3481 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc 3541 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag 3601 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac 3661 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa 3721 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa 3781 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta 3841 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc 3901 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag 3961 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc 4021 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt 4081 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg 4141 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc 4201 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt 4261 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag 4321 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc 4381 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc 4441 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga 4501 acggatctcg acggtatcgc caaatggcag tattcatcca caattttaaa agaaaagggg 4561 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 4621 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 4681 gcagagatcc agtttggatc gataagcttg atatcgaatt caggcaggca gaagtatgca 4741 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 4801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 4861 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 4921 tggaggccta ggctacgcgc gccaccatgc ccaagaagaa gcgcaaggtg ggacgcgtct 4981 gcaggatatc aagcttgcgg taccgcgggc ccgggatcgc caccatggac aagaagtaca 5041 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca 5101 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga 5161 acctgatcgg cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga 5221 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct 5281 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc 5341 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg 5401 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca 5461 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg 5521 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca 5581 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg 5641 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga 5701 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc 5761 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc 5821 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt 5881 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc 5941 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg 6001 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga 6061 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg 6121 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca 6181 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg 6241 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc 6301 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct 6361 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga 6421 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg 6481 ccagcgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga 6541 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca 6601 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa 6661 aagccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag 6721 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc 6781 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact 6841 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt 6901 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca 6961 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc 7021 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg 7081 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag 7141 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca 7201 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg 7261 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag 7321 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag 7381 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc 7441 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc 7501 aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct 7561 ttctgaagga cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca 7621 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc 7681 tgctgaatgc caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag 7741 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc 7801 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga 7861 acgacaaact gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt 7921 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg 7981 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa 8041 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg 8101 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt 8161 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa 8221 acggcgaaac aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag 8281 tgctgtctat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca 8341 gcaaagagtc tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact 8401 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg 8461 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga 8521 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg 8581 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc 8641 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac 8701 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga 8761 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc 8821 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta 8881 atctggacaa ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg 8941 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt 9001 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca 9061 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg 9121 gaggcgacgc ctatccctat gacgtgcccg attatgccag cctgggcagc ggctccccca 9181 agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacggc attggtagtg 9241 ggagcaacgg cagcagcgga tccaacggtc cgggtggatc tggaggtgga ggttctggag 9301 gagaagaact tttgagcaag aattatcatc ttgagaacga agtggctcgt cttaagaaag 9361 gttctggcag tggagaagaa ctgctttcaa agaattacca cctggaaaat gaggtagcta 9421 gactgaaaaa ggggagcgga agtggggagg agttgctgag caaaaattat catttggaga 9481 acgaagtagc acgactaaag aaagggtccg gatcgggtga ggagttactc tcgaaaaatt 9541 atcatctcga aaacgaagtg gctcggctaa aaaagggcag tggttctgga gaagagctat 9601 tatctaaaaa ctaccacctc gaaaatgagg tggcacgctt aaaaaaggga agtggcagtg 9661 gtgaagagct actatccaag aattatcatc ttgagaacga ggtagcgcgt ttgaagaagg 9721 gttccggctc aggagaggaa ctgctctcga agaactatca tcttgaaaat gaggtcgctc 9781 gattaaaaaa gggatcgggc agtggtgagg aactactttc aaagaattac cacctcgaaa 9841 acgaagtagc tcgattaaag aaaggttcag ggtcgggtga agaattactg agtaaaaatt 9901 atcatctgga aaatgaggta gcgagactaa aaaaggggag tggttctggc gaagagttgc 9961 tatcgaaaaa ttatcatctt gagaacgaag ttgctaggct caaaaagggc tcaggctcag 10021 gcgaggagtt gctctcgaaa aactaccact tggaaaatga ggtcgcgagg ttgaaaaagg 10081 ggagcgggtc gggcgaggag ttattgagca aaaactatca tttagagaac gaagtcgcgc 10141 gcttaaagaa aggctcgggc tcgggcgaag aactcttatc gaagaactac cacctcgaaa 10201 atgaggtcgc caggttgaaa aagggcagtg gcagcgggga ggaactcttg agcaagaact 10261 accacttgga gaatgaggtc gcgagattga agaaagggtc ggggagcggc gaggaattgc 10321 tcagcaagaa ttatcatttg gagaacgaag tcgccaggct caagaaaggc tcggggtcgg 10381 gggaggaatt gttgagtaaa aactaccact tggaaaatga agtcgccagg ctcaaaaaag 10441 ggagtgggag cggcgaagag ttattgagca aaaattacca cttggagaac gaagtggcaa 10501 ggctcaagaa agggagcggc agcggggagg agctcttatc gaagaactac cacttagaga 10561 atgaagtcgc ccgcttgaag aaaggctcgg ggagcgggga agagctcttg agcaagaact 10621 accacttgga aaatgaggtg gcgcgcttga agaaagggag cgggagcggg gaagagttac 10681 tatctaagaa ttatcatctc gagaacgagg tggctcgact aaagaagggc tccggcagtg 10741 gggaggaact cctgtcgaag aactatcatc ttgaaaatga ggttgcaaga cttaaaaagg 10801 ggtccggatc aggtgaggaa ctactcagta agaattacca cctggaaaac gaagttgcac 10861 gtttgaagaa aggatcagga tcaggcgaag aactgctctc aaaagattat catttggaaa 10921 atgaggttgc acgtttaaaa aagggaagtg gcagtggtga ggaacttctg tcgaaaaatt 10981 atcatctcga gaatgaagta gcccgactta aaaagggttc tggctcgggt cagcggccgc 11041 aaggtggagg tggacccaag aagaagcgca aggtgggaag cggagctact aacttcagcc 11101 tgctgaagca ggctggagac gtggaggaga accctggacc tgcaccagga agtatgagcg 11161 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc 11221 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa 11281 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc 11341 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt 11401 ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga 11461 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag 11521 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct 11581 tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga 11641 agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac 11701 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca 11761 aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg 11821 acctccctag caaactgggg cacaagctta attaagcggc cgcgactcta gagtcgacct 11881 gcagggatcg atcactgagg tacctttaag accaatgact tacaaggcag ctgtagatct 11941 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca 12001 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct 12061 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 12121 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 12181 gtcagtgtgg aaaatctcta gcagcatcta gaattaattc cgtgtattct atagtgtcac 12241 ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg ttctaacgac 12301 aatatgtaca agcctaattg tgtagcatct ggcttactga agcagaccct atcatctctc 12361 tcgtaaactg ccgtcagagt cggtttggtt ggacgaacct tctgagtttc tggtaacgcc 12421 gtcccgcacc cggaaatggt cagcgaacca atcagcaggg tcatcgctag ccagatcctc 12481 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat 12541 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt 12601 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg 12661 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc 12721 ctaatgcagg agtcgcataa gggagagcgt cgaatggtgc actctcagta caatctgctc 12781 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 12841 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 12901 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg 12961 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 13021 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 13081 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 13141 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 13201 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 13261 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 13321 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 13381 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 13441 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 13501 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 13561 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 13621 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 13681 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 13741 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 13801 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 13861 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 13921 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 13981 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 14041 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 14101 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 14161 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 14221 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 14281 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 14341 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 14401 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 14461 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 14521 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 14581 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 14641 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 14701 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 14761 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 14821 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 14881 taccgctcgc cgcagccgaa cgaccgagcg cagc //