LOCUS Exported 13968 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses an intron-containing dCas9-VPR fusion driven by human SYN promoter. ACCESSION . VERSION . KEYWORDS lenti SYN-SVI-dCas9-VPR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13968) TITLE A Cre-dependent CRISPR/dCas9 activation system for gene expression regulation in neurons JOURNAL Unpublished REFERENCE 2 (bases 1 to 13968) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13968 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(166..1026) /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 789..808 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1027..1131) /gene="bla" /label=AmpR promoter primer_bind complement(1206..1225) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 1397..1776 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1778..1976 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1931..1951 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 1994..2174 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2221..2346 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2839..3072 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3257..3301 /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 3599..3716 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 3761..4208 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 4215..4224 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4224..4289 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 4296..4316 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" intron 5730..5826 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 6040..8538 /codon_start=1 /product="C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" /label=Cas9(C) /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL IHQSITGLYETRIDLSQLGGD" CDS 8539..8586 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 8614..8763 /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 8788..8808 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9238..9594 /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 9613..10182 /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" misc_feature 10219..10807 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(10272..10292) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(10690..10701) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10879..11112 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(11138..11155) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 11144..11368 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11414..11842 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11501..11520) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11711..11732 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(11851..11871) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 11856..12185 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 12036..12171 /label=SV40 ori /note="SV40 origin of replication" primer_bind 12098..12117 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 12233..12280 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 12299..12673 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 12803..12924 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12840..12859) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12894..12913 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(12973..12989) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12973..12989) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(12986..13008) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 12997..13013 /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(13021..13051) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 13066..13087 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(13204..13221) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(13375..13963) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(13455..13474) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 61 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 1021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 1081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 1141 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact 1201 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg 1261 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc 1321 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg 1381 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg 1441 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 1501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 1561 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 1621 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 1681 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 1741 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 1801 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 1861 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 1921 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg 1981 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 2041 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 2101 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 2161 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag 2221 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga 2281 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag 2341 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca 2401 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga 2461 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag 2521 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca 2581 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag 2641 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag 2701 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt 2761 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga 2821 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac 2881 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt 2941 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac 3001 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga 3061 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc 3121 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat 3181 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc 3241 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt 3301 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt 3361 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag 3421 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag 3481 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt 3541 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt 3601 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag 3661 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg 3721 tttattacag ggacagcaga gatccagttt ggttaattaa agtgcaagtg ggttttagga 3781 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc 3841 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga 3901 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc 3961 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc 4021 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg 4081 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg 4141 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga 4201 gagcgcagac cggtgccacc atggactata aggaccacga cggagactac aaggatcatg 4261 atattgatta caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta 4321 tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggccatc ggcaccaact 4381 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc 4441 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca 4501 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac 4561 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg 4621 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc 4681 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca 4741 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct 4801 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga 4861 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc 4921 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca 4981 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga 5041 atggcctgtt cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca 5101 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc 5161 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga 5221 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg 5281 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc 5341 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga 5401 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt 5461 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca 5521 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc 5581 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg 5641 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc 5701 tggccagggg aaacagcaga ttcgcctggg taagtttagt ctttttgtct tttatttcag 5761 gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac 5821 ttctagatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg 5881 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg 5941 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac 6001 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc 6061 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag 6121 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc 6181 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag 6241 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc 6301 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg 6361 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg 6421 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc 6481 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg 6541 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag 6601 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag 6661 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa 6721 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag 6781 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa 6841 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg 6901 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg 6961 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag 7021 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac 7081 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag 7141 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg 7201 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag 7261 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg 7321 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac 7381 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct 7441 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc 7501 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc 7561 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg 7621 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc 7681 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca 7741 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga 7801 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct 7861 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag 7921 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg 7981 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc 8041 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag 8101 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat 8161 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac 8221 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg 8281 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc 8341 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc 8401 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg 8461 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg 8521 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggaca ggccaaaaag 8581 aaaaagctcg agggcggagg cgggagcgga tccgacgcat tggacgattt tgatctggat 8641 atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt 8701 gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg 8761 ctgattaact ctagaagttc cggatctccg aaaaagaaac gcaaagttgg tagccagtac 8821 ctgcccgaca ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac ctacgagaca 8881 ttcaagagca tcatgaagaa gtcccccttc agcggcccca ccgaccctag acctccacct 8941 agaagaatcg ccgtgcccag cagatccagc gccagcgtgc caaaacctgc cccccagcct 9001 taccccttca ccagcagcct gagcaccatc aactacgacg agttccctac catggtgttc 9061 cccagcggcc agatctctca ggcctctgct ctggctccag cccctcctca ggtgctgcct 9121 caggctcctg ctcctgcacc agctccagcc atggtgtctg cactggctca ggcaccagca 9181 cccgtgcctg tgctggctcc tggacctcca caggctgtgg ctccaccagc ccctaaacct 9241 acacaggccg gcgagggcac actgtctgaa gctctgctgc agctgcagtt cgacgacgag 9301 gatctgggag ccctgctggg aaacagcacc gatcctgccg tgttcaccga cctggccagc 9361 gtggacaaca gcgagttcca gcagctgctg aaccagggca tccctgtggc ccctcacacc 9421 accgagccca tgctgatgga ataccccgag gccatcaccc ggctcgtgac aggcgctcag 9481 aggcctcctg atccagctcc tgcccctctg ggagcaccag gcctgcctaa tggactgctg 9541 tctggcgacg aggacttcag ctctatcgcc gatatggatt tctcagcctt gctgggctct 9601 ggcagcggca gccgggattc cagggaaggg atgtttttgc cgaagcctga ggccggctcc 9661 gctattagtg acgtgtttga gggccgcgag gtgtgccagc caaaacgaat ccggccattt 9721 catcctccag gaagtccatg ggccaaccgc ccactccccg ccagcctcgc accaacacca 9781 accggtccag tacatgagcc agtcgggtca ctgaccccgg caccagtccc tcagccactg 9841 gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc cgatgaagag 9901 acgagccagg ctgtcaaagc ccttcgggag atggccgata ctgtgattcc ccagaaggaa 9961 gaggctgcaa tctgtggcca aatggacctt tcccatccgc ccccaagggg ccatctggat 10021 gagctgacaa ccacacttga gtccatgacc gaggatctga acctggactc acccctgacc 10081 ccggaattga acgagattct ggataccttc ctgaacgacg agtgcctctt gcatgccatg 10141 catatcagca caggactgtc catcttcgac acatctctgt tttgagaatt caattctaac 10201 tagacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 10261 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 10321 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 10381 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 10441 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 10501 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 10561 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 10621 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 10681 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 10741 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 10801 tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt 10861 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt 10921 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac 10981 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg 11041 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga 11101 aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 11161 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 11221 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 11281 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 11341 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 11401 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 11461 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 11521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 11581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 11641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 11701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 11761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 11821 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 11881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 11941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 12001 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 12061 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 12121 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 12181 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa 12241 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat 12301 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga 12361 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt 12421 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa 12481 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt 12541 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc 12601 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga 12661 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt 12721 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 12781 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 12841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 12901 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 12961 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 13021 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 13081 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 13141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 13201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 13261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 13321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 13381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 13441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 13501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 13561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 13621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 13681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 13741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 13801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 13861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 13921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaaga //