LOCUS Exported 12983 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION dCas9 fused to BFP and the human DNMT3A catalytically inactive domain. ACCESSION . VERSION . KEYWORDS pCCL-PGK-SPdCas9-BFP-DNMT3A(E752A) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12983) AUTHORS Lin L, Liu Y, Xu F, Huang J, Daugaard TF, Petersen TS, Hansen B, Ye L, Zhou Q, Fang F, Yang L, Li S, Floe L, Jensen KT, Shrock E, Chen F, Yang H, Wang J, Liu X, Xu X, Bolund L, Nielsen AL, Luo Y TITLE Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. JOURNAL Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. PUBMED 29635374 REFERENCE 2 (bases 1 to 12983) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12983 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 82..461 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 462..665 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 616..636 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 680..860 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 904..1029 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1522..1755 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1939..1983 /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 2250..2367 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2406..2905 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(2441..2464) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 2838..2857 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 2964..2973 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2970..7073 /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 7077..7103 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 7122..7142 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7149..7169 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7209..7904 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" misc_feature 8910..9498 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8963..8983) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9381..9392) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 9587..9820 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(9849..9867) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(9850..9867) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(10125..10144) /label=T7 /note="T7 promoter, forward primer" promoter complement(10126..10144) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(10154..10171) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(10154..10170) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10163..10185) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 10315..10770 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10402..10421) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10612..10633 /label=F1ori-F /note="F1 origin, forward primer" promoter 10799..10903 /gene="bla" /label=AmpR promoter CDS 10904..11764 /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(11122..11141) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 11935..12523 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 12424..12443 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 12677..12694 /label=L4440 /note="L4440 vector, forward primer" protein_bind 12811..12832 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 12847..12877 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12885..12901 /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)." primer_bind 12890..12912 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 12909..12925 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 12909..12925 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 12944..12964 /label=T3 /note="T3 promoter, forward primer" promoter 12946..12964 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 aagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat 61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 661 gagctcgttt agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc 721 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 781 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 841 agtgtggaaa atctctagca gtggcgcccg aacagggacc tgaaagcgaa agggaaacca 901 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg 961 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc 1021 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg 1081 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa 1141 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga 1201 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta 1261 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac 1321 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt 1381 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt 1441 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag 1501 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag 1561 cactatgggc gcagcctcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat 1621 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact 1681 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa 1741 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt 1801 gccttggaat gctagttgga gtaataaatc tctggaacag attggaatca cacgacctgg 1861 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa 1921 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt 1981 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata 2041 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt 2101 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac 2161 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta 2221 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac 2281 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa 2341 aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga cgatggtcga 2401 gtaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc 2461 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg 2521 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct 2581 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt 2641 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg 2701 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg 2761 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag 2821 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc 2881 ctcttcctca tctccgggcc tttcgacctc tagcgggatc cggcgcgtct gcaggatatc 2941 aagcttgcgg taccgcgggc ccggccacca tggacaagaa gtacagcatc ggcctggcca 3001 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga 3061 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggcgccc 3121 tgctgttcga cagcggagaa acagccgagg ccacccggct gaagagaacc gccagaagaa 3181 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg 3241 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata 3301 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga 3361 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc 3421 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg 3481 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga 3541 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca 3601 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg 3661 gcgagaagaa gaatggcctg ttcggcaacc tgattgccct gagcctgggc ctgaccccca 3721 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct 3781 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc 3841 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg 3901 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg 3961 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt 4021 tcttcgacca gagcaagaac ggctacgccg gctacatcga tggcggagcc agccaggaag 4081 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg 4141 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc 4201 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc 4261 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact 4321 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg 4381 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgccagc gcccagagct 4441 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc 4501 acagcctgct gtacgagtac ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga 4561 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaagcc atcgtggacc 4621 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga 4681 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc 4741 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg 4801 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga 4861 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc 4921 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc 4981 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca 5041 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag 5101 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc 5161 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga 5221 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc 5281 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc 5341 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc 5401 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca 5461 accggctgtc cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact 5521 ccatcgataa caaagtgctg actcggagcg acaagaaccg gggcaagagc gacaacgtgc 5581 cctccgaaga ggtcgtgaag aagatgaaga actactggcg ccagctgctg aatgccaagc 5641 tgattaccca gaggaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac 5701 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg 5761 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaacgac aaactgatcc 5821 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc 5881 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg 5941 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg 6001 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca 6061 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta 6121 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaacaggcg 6181 agatcgtgtg ggataagggc cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc 6241 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc 6301 tgcccaagag gaacagcgac aagctgatcg ccagaaagaa ggactgggac cctaagaagt 6361 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa 6421 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa 6481 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga 6541 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga 6601 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca 6661 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg 6721 ataatgagca gaaacagctg tttgtggaac agcacaaaca ctacctggac gagatcatcg 6781 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaggtgc 6841 tgagcgccta caacaagcac agagacaagc ctatcagaga gcaggccgag aatatcatcc 6901 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca 6961 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga 7021 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc gacgcctatc 7081 cctatgacgt gcccgattat gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg 7141 tggaagatcc taagaaaaag cggaaagtgg acggcattgg tagtgggagc aacggcagca 7201 gcggatccag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg 7261 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc 7321 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg 7381 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact 7441 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg 7501 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg 7561 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg 7621 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg 7681 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata 7741 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac 7801 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg 7861 ccagatactg cgacctccct agcaaactgg ggcacaagct taatggtgga ggaggttctg 7921 gaggcggtgg aagtggtggc ggaggtagcg tcgacaacca cgaccaggaa tttgaccctc 7981 caaaggttta cccacctgtc ccagctgaga agaggaagcc catccgggtg ctgtctctct 8041 ttgatggaat cgctacaggg ctcctggtgc tgaaggactt gggcattcag gtggaccgct 8101 acattgcctc ggaggtgtgt gaggactcca tcacggtggg catggtgcgg caccagggga 8161 agatcatgta cgtcggggac gtccgcagcg tcacacagaa gcatatccag gagtggggcc 8221 cattcgatct ggtgattggg ggcagtccct gcaatgacct ctccatcgtc aaccctgctc 8281 gcaagggcct ctacgagggc actggccggc tcttctttga gttctaccgc ctcctgcatg 8341 atgcgcggcc caaggaggga gatgatcgcc ccttcttctg gctctttgcc aatgtggtgg 8401 ccatgggcgt tagtgacaag agggacatct cgcgatttct cgagtccaac cctgtgatga 8461 ttgatgccaa agaagtgtca gctgcacaca gggcccgcta cttctggggt aaccttcccg 8521 gtatgaacag gccgttggca tccactgtga atgataagct ggagctgcag gagtgtctgg 8581 agcatggcag gatagccaag ttcagcaaag tgaggaccat tactacgagg tcaaactcca 8641 taaagcaggg caaagaccag cattttcctg tcttcatgaa tgagaaagag gacatcttat 8701 ggtgcactga aatggaaagg gtatttggtt tcccagtcca ctatactgac gtctccaaca 8761 tgagccgctt ggcgaggcag agactgctgg gccggtcatg gagcgtgcca gtcatccgcc 8821 acctcttcgc tccgctgaag gagtattttg cgtgtgtgta atagatgcat ggccggcctt 8881 aattaaacgc gtctcgaggg aattccgata atcaacctct ggattacaaa atttgtgaaa 8941 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 9001 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 9061 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 9121 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 9181 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 9241 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 9301 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 9361 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 9421 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 9481 tttgggccgc ctccccgcat cgggaattcg agctcggtac ctttaagacc aatgacttac 9541 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 9601 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 9661 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 9721 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 9781 tccctcagac ccttttagtc agtgtggaaa atctctagca gcatctagct agaattaatt 9841 ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa 9901 ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg 9961 aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc 10021 ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg 10081 gtcatcgcta gcctaggctt ttgcgtcgag acgtacccaa ttcgccctat agtgagtcgt 10141 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta 10201 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg 10261 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc 10321 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 10381 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 10441 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 10501 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 10561 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 10621 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 10681 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 10741 attttaacaa aatattaacg tttacaattt cccaggtggc acttttcggg gaaatgtgcg 10801 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 10861 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 10921 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 10981 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 11041 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 11101 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca 11161 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 11221 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 11281 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 11341 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 11401 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac 11461 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 11521 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 11581 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 11641 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 11701 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 11761 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 11821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 11881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 11941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 12001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 12061 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 12121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 12181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 12241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 12301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 12361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 12421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 12481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 12541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 12601 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 12661 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa 12721 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga 12781 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc 12841 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca 12901 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta 12961 aagggaacaa aagctggagc tgc //