LOCUS Exported 13871 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentivirus compatible dCas9-VPR construct driven by the human SYN promoter. ACCESSION . VERSION . KEYWORDS lenti SYN-FLAG-dCas9-VPR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13871) AUTHORS Savell KE, Bach SV, Zipperly ME, Revanna JS, Goska NA, Tuscher JJ, Duke CG, Sultan FA, Burke JN, Williams D, Ianov L, Day JJ TITLE A Neuron-Optimized CRISPR/dCas9 Activation System for Robust and Specific Gene Regulation. JOURNAL eNeuro. 2019 Mar 7;6(1). pii: eN-MNT-0495-18. doi: 10.1523/ENEURO.0495-18.2019. eCollection 2019 Jan-Feb. PUBMED 30863790 REFERENCE 2 (bases 1 to 13871) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13871 /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" CDS 4341..8441 /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 8442..8489 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 8517..8666 /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 8691..8711 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9141..9497 /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 9516..10085 /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 10122..10710 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(10175..10195) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(10593..10604) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10782..11015 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(11041..11058) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 11047..11271 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11317..11745 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11404..11423) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11614..11635 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(11754..11774) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 11759..12088 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 11939..12074 /label=SV40 ori /note="SV40 origin of replication" primer_bind 12001..12020 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 12136..12183 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 12202..12576 /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 12706..12827 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12743..12762) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12797..12816 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(12876..12892) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12876..12892) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(12889..12911) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 12900..12916 /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(12924..12954) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12969..12990 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(13107..13124) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(13278..13866) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(13358..13377) /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 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc 5761 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga 5821 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt 5881 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga 5941 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga 6001 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct 6061 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc 6121 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca 6181 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac 6241 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga 6301 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt 6361 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc 6421 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg 6481 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga 6541 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca 6601 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga 6661 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga 6721 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact 6781 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg 6841 actacgatgt ggacgctatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca 6901 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg 6961 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga 7021 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg 7081 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc 7141 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag 7201 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag 7261 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa 7321 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg 7381 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca 7441 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg 7501 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg 7561 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg 7621 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga 7681 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg 7741 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca 7801 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg 7861 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga 7921 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg 7981 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact 8041 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga 8101 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg 8161 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca 8221 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc 8281 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga 8341 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc 8401 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga 8461 aaaaggccgg acaggccaaa aagaaaaagc tcgagggcgg aggcgggagc ggatccgacg 8521 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg 8581 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc 8641 ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct ccgaaaaaga 8701 aacgcaaagt tggtagccag tacctgcccg acaccgacga ccggcaccgg atcgaggaaa 8761 agcggaagcg gacctacgag acattcaaga gcatcatgaa gaagtccccc ttcagcggcc 8821 ccaccgaccc tagacctcca cctagaagaa tcgccgtgcc cagcagatcc agcgccagcg 8881 tgccaaaacc tgccccccag ccttacccct tcaccagcag cctgagcacc atcaactacg 8941 acgagttccc taccatggtg ttccccagcg gccagatctc tcaggcctct gctctggctc 9001 cagcccctcc tcaggtgctg cctcaggctc ctgctcctgc accagctcca gccatggtgt 9061 ctgcactggc tcaggcacca gcacccgtgc ctgtgctggc tcctggacct ccacaggctg 9121 tggctccacc agcccctaaa cctacacagg ccggcgaggg cacactgtct gaagctctgc 9181 tgcagctgca gttcgacgac gaggatctgg gagccctgct gggaaacagc accgatcctg 9241 ccgtgttcac cgacctggcc agcgtggaca acagcgagtt ccagcagctg ctgaaccagg 9301 gcatccctgt ggcccctcac accaccgagc ccatgctgat ggaatacccc gaggccatca 9361 cccggctcgt gacaggcgct cagaggcctc ctgatccagc tcctgcccct ctgggagcac 9421 caggcctgcc taatggactg ctgtctggcg acgaggactt cagctctatc gccgatatgg 9481 atttctcagc cttgctgggc tctggcagcg gcagccggga ttccagggaa gggatgtttt 9541 tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt tgagggccgc gaggtgtgcc 9601 agccaaaacg aatccggcca tttcatcctc caggaagtcc atgggccaac cgcccactcc 9661 ccgccagcct cgcaccaaca ccaaccggtc cagtacatga gccagtcggg tcactgaccc 9721 cggcaccagt ccctcagcca ctggatccag cgcccgcagt gactcccgag gccagtcacc 9781 tgttggagga tcccgatgaa gagacgagcc aggctgtcaa agcccttcgg gagatggccg 9841 atactgtgat tccccagaag gaagaggctg caatctgtgg ccaaatggac ctttcccatc 9901 cgcccccaag gggccatctg gatgagctga caaccacact tgagtccatg accgaggatc 9961 tgaacctgga ctcacccctg accccggaat tgaacgagat tctggatacc ttcctgaacg 10021 acgagtgcct cttgcatgcc atgcatatca gcacaggact gtccatcttc gacacatctc 10081 tgttttgaga attcaattct aactagacgc gttaagtcga caatcaacct ctggattaca 10141 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 10201 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 10261 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 10321 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 10381 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 10441 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 10501 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga 10561 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 10621 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 10681 gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc 10741 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc 10801 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg 10861 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 10921 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 10981 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag 11041 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 11101 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 11161 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 11221 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 11281 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 11341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 11401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 11461 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 11521 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 11581 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 11641 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 11701 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 11761 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 11821 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 11881 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 11941 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 12001 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 12061 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 12121 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac 12181 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg 12241 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc 12301 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc 12361 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag 12421 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc 12481 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc 12541 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc 12601 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 12661 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 12721 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 12781 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 12841 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 12901 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 12961 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 13021 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 13081 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 13141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 13201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 13261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 13321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 13381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 13441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 13501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 13561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 13621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 13681 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 13741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 13801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 13861 tctcaagaag a //