LOCUS Exported 10512 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ST1-dCas9 with VP64-p65-Rta (VPR) fused to it's C-terminus; mammalian vector. ACCESSION . VERSION . KEYWORDS M_ST1n_VPR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10512) AUTHORS Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan D, Braff JL, Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ, Church GM TITLE Highly efficient Cas9-mediated transcriptional programming. JOURNAL Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312. PUBMED 25730490 REFERENCE 2 (bases 1 to 10512) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10512 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 50..429 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 430..633 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 584..604 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 630..654 /label=LNCX /note="Human CMV promoter, forward primer" CDS 748..4110 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system" /label=St1dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A" /translation="MSDLVLGLAIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC LYTGKTISIHDLINNSNQFEVAAILPLSITFDDSLANKVLVYATAAQEKGQRTPYQALD SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTRYASRVV LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ HIIKNEGDKPKLDF" CDS 4123..4143 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4168..4317 /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 4342..4362 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4792..5148 /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 5167..5736 /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" protein_bind complement(5741..5765) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter complement(5815..5832) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5898..5917) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5942..5990 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6192..6620 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6279..6298) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6489..6510 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6629..6649) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6634..6963 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6814..6949 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6876..6895 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7030..7824 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7084..7103) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7694..7713 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 8000..8121 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8037..8056) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8091..8110 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8170..8186) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8170..8186) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8183..8205) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8194..8210 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(8218..8248) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8263..8284 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8401..8418) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8572..9160) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8652..8671) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9331..10191) /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 9954..9973 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10192..10296) /gene="bla" /label=AmpR promoter primer_bind complement(10371..10390) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagg atcgaaccct tgccaccatg agcgacctgg tgctgggcct ggccatcggc 781 atcggcagcg tgggcgtggg catcctgaac aaggtgaccg gcgagatcat ccacaagaac 841 agtcgcatct tccctgctgc tcaggctgag aacaacctgg tgcgccgcac caaccgccag 901 ggtcgccggc ttgctcgccg caagaagcac cggcgcgtgc gcctgaaccg cctgttcgag 961 gagagcggcc tgatcaccga cttcaccaag atcagcatca acctgaaccc ctaccagctg 1021 cgcgtgaagg gcctgaccga cgagctgagc aacgaggagc tgttcatcgc cctgaagaac 1081 atggtgaagc accgcggcat cagctacctg gacgacgcca gcgacgacgg caacagcagc 1141 gtgggcgact acgcccagat cgtgaaggag aacagcaagc agctggagac caagaccccc 1201 ggccagatcc agctggagcg ctaccagacc tacggccagc tgcgcggcga cttcaccgtg 1261 gagaaggacg gcaagaagca ccgcctgatc aacgtgttcc ccaccagcgc ctaccgcagc 1321 gaggccctgc gcatcctgca gacccagcag gagttcaacc cccagatcac cgacgagttc 1381 atcaaccgct acctggagat cctgaccggc aagcgcaagt actaccacgg ccccggcaac 1441 gagaagagcc gcaccgacta cggccgctac cgcaccagcg gcgagaccct ggacaacatc 1501 ttcggcatcc tgatcggcaa gtgcaccttc taccccgacg agttccgcgc cgccaaggcc 1561 agctacaccg cccaggagtt caacctgctg aacgacctga acaacctgac cgtgcccacc 1621 gagaccaaga agctgagcaa ggagcagaag aaccagatca tcaactacgt gaagaacgag 1681 aaggccatgg gccccgccaa gctgttcaag tacatcgcca agctgctgag ctgcgacgtg 1741 gccgacatca agggctaccg catcgacaag agcggcaagg ccgagatcca caccttcgag 1801 gcctaccgca agatgaagac cctggagacc ctggacatcg agcagatgga ccgcgagacc 1861 ctggacaagc tggcctacgt gctgaccctg aacaccgagc gcgagggcat ccaggaggcc 1921 ctggagcacg agttcgccga cggcagcttc agccagaagc aggtggacga gctggtgcag 1981 ttccgcaagg ccaacagcag catcttcggc aagggctggc acaacttcag cgtgaagctg 2041 atgatggagc tgatccccga gctgtacgag accagcgagg agcagatgac catcctgacc 2101 cgcctgggca agcagaagac caccagcagc agcaacaaga ccaagtacat cgacgagaag 2161 ctgctgaccg aggagatcta caaccccgtg gtggccaaga gcgtgcgcca ggccatcaag 2221 atcgtgaacg ccgccatcaa ggagtacggc gacttcgaca acatcgtgat cgagatggcc 2281 cgcgagacca acgaggacga cgagaagaag gccatccaga agatccagaa ggccaacaag 2341 gacgagaagg acgccgccat gctgaaggcc gccaaccagt acaacggcaa ggccgagctg 2401 ccccacagcg tgttccacgg ccacaagcag ctggccacca agatccgcct gtggcaccag 2461 cagggcgagc gctgcctgta caccggcaag accatcagca tccacgacct gatcaacaac 2521 agcaaccagt tcgaggtggc tgccatcctg cccctgagca tcaccttcga cgacagcctg 2581 gccaacaagg tgctggtgta cgccaccgcc gctcaggaga agggccagcg caccccctac 2641 caggccctgg acagcatgga cgacgcctgg agcttccgcg agctgaaggc cttcgtgcgc 2701 gagagcaaga ccctgagcaa caagaagaag gagtacctgc tgaccgagga ggacatcagc 2761 aagttcgacg tgcgcaagaa gttcatcgag cgcaacctgg tggacacccg ctacgccagc 2821 cgcgtggtgc tgaacgccct gcaggagcac ttccgcgccc acaagatcga caccaaggtg 2881 agcgtggtgc gcggccagtt caccagccag ctgcgccgcc actggggcat cgagaagacc 2941 cgcgacacct accaccacca cgccgtggac gccctgatca ttgcggcttc tagccagctg 3001 aacctgtgga agaagcagaa gaacaccctg gtgagctaca gcgaggacca gctgctggac 3061 atcgagaccg gcgagctgat cagcgacgac gagtacaagg agagcgtgtt caaggccccc 3121 taccagcact tcgtggacac cctgaagagc aaggagttcg aggacagcat cctgttcagc 3181 taccaggtgg acagcaagtt caaccgcaag atcagcgacg ccaccatcta cgccacccgc 3241 caggccaagg tgggcaagga caaggccgac gagacctacg tgctgggcaa gatcaaggac 3301 atctacaccc aggacggcta cgacgccttc atgaagatct acaagaagga caagagcaag 3361 ttcctgatgt accgccacga cccccagacc ttcgagaagg tgatcgagcc catcctggag 3421 aactacccca acaagcagat caacgataaa ggcaaggagg tgccctgcaa ccccttcctg 3481 aagtacaagg aggagcacgg ctacatccgc aagtacagca agaagggcaa cggccccgag 3541 atcaagagcc tgaagtacta cgacagcaag ctgggcaacc acatcgacat cacccccaag 3601 gacagcaaca acaaggtggt gctgcagagc gtgagcccct ggcgcgccga cgtgtacttc 3661 aacaagacca ccggcaagta cgagatcctg ggcctgaagt acgccgacct gcagtttgat 3721 aagggcaccg gcacctacaa gatcagccag gagaagtaca acgacatcaa gaagaaggag 3781 ggcgtggaca gcgacagcga gttcaagttc accctgtaca agaacgacct tctgctggtg 3841 aaggacaccg agaccaagga gcaacagctg ttccgcttcc tgagccgcac catgcccaag 3901 cagaagcact acgtggagct gaagccctac gacaagcaga agttcgaggg cggcgaggcc 3961 ctgatcaagg tgctgggcaa cgtggccaac agcggccagt gcaagaaggg cctgggcaag 4021 agcaacatca gcatctacaa ggtgcgcacc gacgtgctgg gcaaccagca catcatcaag 4081 aacgagggcg acaagcccaa gctggacttc agcagggctg accccaagaa gaagaggaag 4141 gtggaggcca gcggttccgg acgggctgac gcattggacg attttgatct ggatatgctg 4201 ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac 4261 tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catgctgatt 4321 aactctagaa gttccggatc tccgaaaaag aaacgcaaag ttggtagcca gtacctgccc 4381 gacaccgacg accggcaccg gatcgaggaa aagcggaagc ggacctacga gacattcaag 4441 agcatcatga agaagtcccc cttcagcggc cccaccgacc ctagacctcc acctagaaga 4501 atcgccgtgc ccagcagatc cagcgccagc gtgccaaaac ctgcccccca gccttacccc 4561 ttcaccagca gcctgagcac catcaactac gacgagttcc ctaccatggt gttccccagc 4621 ggccagatct ctcaggcctc tgctctggct ccagcccctc ctcaggtgct gcctcaggct 4681 cctgctcctg caccagctcc agccatggtg tctgcactgg ctcaggcacc agcacccgtg 4741 cctgtgctgg ctcctggacc tccacaggct gtggctccac cagcccctaa acctacacag 4801 gccggcgagg gcacactgtc tgaagctctg ctgcagctgc agttcgacga cgaggatctg 4861 ggagccctgc tgggaaacag caccgatcct gccgtgttca ccgacctggc cagcgtggac 4921 aacagcgagt tccagcagct gctgaaccag ggcatccctg tggcccctca caccaccgag 4981 cccatgctga tggaataccc cgaggccatc acccggctcg tgacaggcgc tcagaggcct 5041 cctgatccag ctcctgcccc tctgggagca ccaggcctgc ctaatggact gctgtctggc 5101 gacgaggact tcagctctat cgccgatatg gatttctcag ccttgctggg tagcggatcg 5161 ggatctcggg attccaggga agggatgttt ttgccgaagc ctgaggccgg ctccgctatt 5221 agtgacgtgt ttgagggccg cgaggtgtgc cagccaaaac gaatccggcc atttcatcct 5281 ccaggaagtc catgggccaa ccgcccactc cccgccagcc tcgcaccaac accaaccggt 5341 ccagtacatg agccagtcgg gtcactgacc ccggcaccag tccctcagcc actggatcca 5401 gcgcccgcag tgactcccga ggccagtcac ctgttggagg atcccgatga agagacgagc 5461 caggctgtca aagcccttcg ggagatggcc gatactgtga ttccccagaa ggaagaggct 5521 gcaatctgtg gccaaatgga cctttcccat ccgcccccaa ggggccatct ggatgagctg 5581 acaaccacac ttgagtccat gaccgaggat ctgaacctgg actcacccct gaccccggaa 5641 ttgaacgaga ttctggatac cttcctgaac gacgagtgcc tcttgcatgc catgcatatc 5701 agcacaggac tgtccatctt cgacacatct ctgttttgaa acccagcttt cttgtacaaa 5761 gtggttgata tcatctagag cggccgcagg tacctgaata actaaggccg cttcccttta 5821 gtgagggtta atgcttcgag cagacaaagg gttcgatccc taccggttag taatgagttt 5881 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg 5941 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg 6001 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata 6061 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct 6121 cgcagccaac gtcggggcgg cagaccctgc catagcagat ctgcgcagct ggggctctag 6181 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 6241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 6301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg 6361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 6421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 6481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 6541 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta 6601 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 6661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 6721 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 6781 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6841 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6901 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 6961 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 7021 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 7081 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 7141 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 7201 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 7261 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 7321 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 7381 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 7441 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 7501 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 7561 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 7621 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 7681 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 7741 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 7801 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc 7861 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 7921 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 7981 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 8041 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 8101 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 8161 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 8221 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 8281 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 8341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 8401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 8461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 8521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 8581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 8641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 8701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 8761 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 8821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 8881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 8941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 9001 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 9061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 9121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 9181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 9241 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 9301 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 9361 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 9421 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 9481 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 9541 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 9601 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 9661 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 9721 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 9781 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 9841 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 9901 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 9961 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 10021 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 10081 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 10141 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 10201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 10261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 10321 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc 10381 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag 10441 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag 10501 aatctgctta gg //