LOCUS Exported 13263 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION TU for the constitutive expression of dCas9 fused to VPR (VP64-p65-Rta) activation domains. ACCESSION . VERSION . KEYWORDS pDGB3 alpha2 P35S:dCas9:VPR:Tnos (GB1826) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13263) AUTHORS Selma S, Bernabe-Orts JM, Vazquez-Vilar M, Diego-Martin B, Ajenjo M, Garcia-Carpintero V, Granell A, Orzaez D TITLE Strong gene activation in plants with genome-wide specificity using a new orthogonal CRISPR/Cas9-based programmable transcriptional activator. JOURNAL Plant Biotechnol J. 2019 Sep;17(9):1703-1705. doi: 10.1111/pbi.13138. Epub 2019 May 23. PUBMED 31034138 REFERENCE 2 (bases 1 to 13263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13263 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 637..981 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 925..945 /label=35S promoter /note="CaMV 35S promoter, forward primer" CDS 1058..1078 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1079..5179 /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5198..5218 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5222..5242 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5246..5266 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5267..5416 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 5417..5437 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5441..5461 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5462..5818 /codon_start=1 /product="transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)" /label=RelA (p65) AD /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD EDFSSIADMDFSALL" CDS 5843..6412 /codon_start=1 /product="transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)" /label=Rta AD /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL DTFLNDECLLHAMHISTGLSIFDTSLF" terminator 6439..6691 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(6726..6744) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 6928..6952 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 8253..8882 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 9311..10384 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 10450..10644 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(10880..10902) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 10988..11128 /label=bom /note="basis of mobility region from pBR322" primer_bind 11043..11062 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(11143..11160) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11314..11902) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11394..11413) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11990..12805) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 12713..12732 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" misc_feature 13238..13262 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" ORIGIN 1 taacaagctt cgtctcagtc aggagactag agccaagctg atctcctttg ccccggagat 61 caccatggac gactttctct atctctacga tctaggaaga aagttcgacg gagaaggtga 121 cgataccatg ttcaccaccg ataatgagaa gattagcctc ttcaatttca gaaagaatgc 181 tgacccacag atggttagag aggcctacgc ggcaggtctg atcaagacga tctacccgag 241 taataatctc caggagatca aataccttcc caagaaggtt aaagatgcag tcaaaagatt 301 caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca gaagtactat 361 tccagtatgg acgattcaag gcttgcttca taaaccaagg caagtaatag agattggagt 421 ctctaagaaa gtagttccta ctgaatcaaa ggccatggag tcaaaaattc agatcgagga 481 tctaacagaa ctcgccgtga agactggcga acagttcata cagagtcttt tacgactcaa 541 tgacaagaag aaaatcttcg tcaacatggt ggagcacgac actctcgtct actccaagaa 601 tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat 661 atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt 721 agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca 781 agatgcccct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga 841 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga 901 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag 961 ttcatttcat ttggagagga ctccggtatt tttacaacaa ttaccacaac aaaacaaaca 1021 acaaacaaca ttacaattta ctattctagt cgaaatgccc aagaagaaga ggaaggtgga 1081 caagaagtac tccattgggc tcgctatcgg cacaaacagc gtcggctggg ccgtcattac 1141 ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag 1201 cataaagaag aacctcattg gcgccctcct gttcgactcc ggggaaacgg ccgaagccac 1261 gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct 1321 gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga 1381 ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat 1441 cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct 1501 tgtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat 1561 caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga 1621 caaactcttt atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa 1681 cgcatccgga gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct 1741 cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat 1801 cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc 1861 caagcttcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat 1921 cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct 1981 gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat 2041 caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca 2101 actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata 2161 cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa 2221 aatggacggc accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca 2281 gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat 2341 cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa 2401 aatcctcaca tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt 2461 cgcgtggatg actcgcaaat cagaagagac tatcactccc tggaacttcg aggaagtcgt 2521 ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct 2581 gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa 2641 cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg 2701 agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa 2761 acagctcaaa gaagattatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg 2821 agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa 2881 agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac 2941 ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct 3001 cttcgacgac aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct 3061 gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt 3121 tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct 3181 cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca gtctccacga 3241 gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa 3301 ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga 3361 gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa 3421 gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga 3481 aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat 3541 gtacgtggat caggaactgg acatcaatcg gctctccgac tacgacgtgg atgccatcgt 3601 gccccagtct tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa 3661 aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta 3721 ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa 3781 ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt 3841 tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa 3901 gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct 3961 ggtttcagat ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca 4021 ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc 4081 caagcttgaa tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat 4141 cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat 4201 tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact 4261 tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac 4321 agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac 4381 cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg 4441 caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag 4501 tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga 4561 actgctgggc atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct 4621 cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc 4681 tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa 4741 aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta 4801 tgaaaagctc aaaggatctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca 4861 caaacactac cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct 4921 cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat 4981 cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc 5041 agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt 5101 cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct 5161 ctctcagctc ggtggagaca gcagggctga ttcggaccca aaaaagaagc gtaaggtcga 5221 tcccaagaag aagagaaagg tagatcctaa gaagaagaga aaggtagacg cattggatga 5281 cttcgactta gacatgcttg ggtcagatgc tttggacgac tttgacctcg acatgttggg 5341 ttctgacgcg ctagatgact tcgacttgga catgttgggc tccgacgctt tggacgactt 5401 tgatctggac atgttaccga aaaagaaacg caaagttgat cctaagaaga agagaaaggt 5461 acctacacag gccggcgagg gcacactgtc tgaagctctg ctgcagctgc agttcgacga 5521 cgaggatctg ggagccctgc tgggaaacag caccgatcct gccgtgttca ccgacctggc 5581 cagcgtggac aacagcgagt tccagcagct gctgaaccag ggcatccctg tggcccctca 5641 caccaccgag cccatgctga tggaataccc cgaggccatc acccggctcg tgacaggcgc 5701 tcagaggcct cctgatccag ctcctgcccc tctgggagca ccaggcctgc ctaatggact 5761 gctgtctggc gacgaggact tcagctctat cgccgatatg gatttctcag ccttgctgat 5821 taactctaga agttccggat ctcgggattc cagggaaggg atgtttttgc cgaagcctga 5881 ggccggctcc gctattagtg acgtgtttga gggccgcgag gtgtgccagc caaaacgaat 5941 ccggccattt catcctccag gaagtccatg ggccaaccgc ccactccccg ccagcctcgc 6001 accaacacca accggtccag tacatgagcc agtcgggtca ctgaccccgg caccagtccc 6061 tcagccactg gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc 6121 cgatgaagaa acgagccagg ctgtcaaagc ccttcgggag atggccgata ctgtgattcc 6181 ccagaaggaa gaggctgcaa tctgtggcca aatggacctt tcccatccgc ccccaagggg 6241 ccatctggat gagctgacaa ccacacttga gtccatgacc gaggatctga acctggactc 6301 acccctgacc ccggaattga acgagattct ggataccttc ctgaacgacg agtgcctctt 6361 gcatgccatg catatcagca caggactgtc catcttcgac acatctctgt tttgagcttg 6421 gaatggatct tcgatcccga tcgttcaaac atttggcaat aaagtttctt aagattgaat 6481 cctgttgccg gtcttgcgac gattatcata taatttctgt tgaattacgt taagcatgta 6541 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg 6601 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta 6661 tcgcgcgcgg tgtcatctat gttactagat cgggaattgc caagctaatt cttgaagacg 6721 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 6781 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 6841 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataatg 6901 ggaccgactc gcgcttgaga cgaagcttga caggatatat tggcgggtaa acctaagaga 6961 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg 7021 tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta ctttgatcca 7081 acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt catctgaaaa 7141 cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct tttcctggcg 7201 ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca 7261 ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga 7321 cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt 7381 ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca 7441 cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg 7501 cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc 7561 agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg 7621 cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc 7681 caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca 7741 cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg 7801 cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga 7861 ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc 7921 cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa 7981 ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag 8041 ccgcccgcgc acctctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc 8101 aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa 8161 aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga 8221 tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga 8281 aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg 8341 gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg 8401 tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg 8461 tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact 8521 tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt 8581 acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg 8641 atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg 8701 gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc 8761 agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg 8821 agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt 8881 gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc 8941 gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa 9001 gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg 9061 ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat 9121 gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc 9181 aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc 9241 caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc cccaagcccg 9301 aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg 9361 tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc 9421 agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg 9481 gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc 9541 agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga 9601 cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga 9661 gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga 9721 ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga 9781 agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt 9841 ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt 9901 aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac 9961 ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg 10021 gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa 10081 gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg 10141 ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa 10201 gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg 10261 caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc 10321 tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc 10381 ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg 10441 actctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa 10501 cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga 10561 tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct 10621 taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa 10681 agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc 10741 ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca 10801 agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt 10861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggtgacggtc acagcttgtc 10921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt 10981 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta 11041 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 11101 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct 11161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 11221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 11281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 11341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 11401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 11461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 11521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 11581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 11641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 11701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 11761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 11821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 11881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 11941 gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtgat tagaaaaact 12001 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt 12061 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa 12121 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc 12181 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg 12241 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct 12301 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga 12361 gtcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc 12421 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata 12481 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac 12541 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca 12601 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg 12661 catcgggctt cccatacaat cggtagattg tcgcacctga ttgcccgaca ttatcgcgag 12721 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc cttgagcaag 12781 acgtttcccg ttgaatatgg ctcataacag aacttattat ttccttcctc ttttctacag 12841 tatttaaaga taccccaaga agctaattat aacaagacga actccaattc actgttcctt 12901 gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg 12961 tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc agcaacgctc 13021 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca 13081 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa 13141 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg 13201 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa 13261 aca //