LOCUS Exported 12258 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION double-DHFR-degron controlled expression of dCas9-BFP-KRAB from the CLYBL locus. ACCESSION . VERSION . KEYWORDS pRT029 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12258) AUTHORS Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME, Kampmann M TITLE CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons. JOURNAL Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 10.1016/j.neuron.2019.07.014. PUBMED 31422865 REFERENCE 2 (bases 1 to 12258) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12258 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(395..1255) /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 1018..1037 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" misc_feature 2113..2138 /label=SA /note="splice acceptor site" primer_bind complement(2180..2197) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2186..2410 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 2416..2795 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2797..3074 /label=chicken beta-actin promoter intron 3076..4093 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 4017..4039 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 4101..4120 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 4164..4637 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILER" regulatory 4656..4665 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4662..8765 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 8769..8795 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 8814..8834 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8841..8861 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8901..9596 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" CDS 9642..9827 /codon_start=1 /product="Kruppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)" /label=KRAB /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY QLTKPDVILRLEKGEEP" CDS 9828..9875 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 9876..10352 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="ISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHT WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR" primer_bind complement(10442..10461) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 10507..10562 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(10561..10580) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" rep_origin complement(11894..224) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11974..11993) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 61 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 1021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 1081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 1141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 1201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 1261 tcctttttca atagcaggag aatcatgcca atgggccaat atacattctg acccacagtt 1321 tcataataaa ataaaatggt tgtggttgta agccactatg tttcagagtg gtttgttaca 1381 cagcaataaa taactaatat agtaggcata ccatcaagtc caaagtaggt agagaagaat 1441 gtaaatagca gagcaaaaca gcatgactgg tggctgggag gcttaaaact gggacaggat 1501 cagagtcatg aaagaagtca aagaaatggt tcagaagtaa ggctgagact gacttacaaa 1561 agctgaaagt ccctttaagt tggtgtttgg tgcattggca ggggcaggta tggtgactta 1621 aaagagccat gctcaacaag atcaagcaca acacaatcac gggtcacccc agcagacctt 1681 agcgagtcta gccatttctt tggtggtggt cacagtcatg cttcagccca gtttccactt 1741 ggacaaatgg tacatatttt caatgagatg aaaattaaga tacaatccat gtgctcagag 1801 agtgatcaca gctctgacta aacactgtgc cccaaagtgt tgaggaattg ggaaaaccta 1861 gctgagttag tggtctcttt tctgttacaa taaagctcat aatgaaaatt agccttcttt 1921 gttcttcccc aagtctttct ttctagacga aactactttc aactgtttta acttccttac 1981 tgttaacttc catatttcta gatagtatgg tcagactgtt atttcttgac ttttaaatgt 2041 aagatattat ctactgactt ccttctatgt aagatgagga ttgagctctc ttacccttct 2101 cccagcggcc gcctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg 2161 atagaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2221 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2281 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 2341 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2401 ggctctatgg gtctcgacat tgattattga ctagttatta atagtaatca attacggggt 2461 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 2521 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 2581 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 2641 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 2701 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 2761 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct 2821 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat 2881 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc 2941 ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg 3001 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa 3061 gcgcgcggcg ggcggggagt cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc 3121 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg 3181 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt 3241 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg 3301 ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc 3361 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga 3421 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg 3481 ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct 3541 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg 3601 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg 3661 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg 3721 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta 3781 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg 3841 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa 3901 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca 3961 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt 4021 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt 4081 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa 4141 agaatgtcga cgctagcgcc accatgatca gtctgattgc ggcgttagcg gtagattacg 4201 ttatcggcat ggaaaacgcc atgccgtgga acctgcctgc cgatctcgcc tggtttaaac 4261 gcaacacctt aaataaaccc gtgattatgg gccgccatac ctgggaatca atcggtcgtc 4321 cgttgccagg acgcaaaaat attatcctca gcagtcaacc gagtacggac gatcgcgtaa 4381 cgtgggtgaa gtcggtggat gaagccatcg cggcgtgtgg tgacgtacca gaaatcatgg 4441 tgattggcgg cggtcgcgtt attgaacagt tcttgccaaa agcgcaaaaa ctgtatctga 4501 cgcatatcga cgcagaagtg gaaggcgaca cccatttccc ggattacgag ccggatgact 4561 gggaatcggt attcagcgaa ttccacgatg ctgatgcgca gaactctcac agctattgct 4621 ttgagattct ggagcggggc tctggaggga ctagtgccac catggacaag aagtacagca 4681 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg 4741 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc 4801 tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa 4861 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca 4921 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg 4981 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg 5041 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg 5101 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc 5161 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc 5221 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg 5281 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg 5341 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg 5401 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga 5461 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg 5521 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga 5581 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg 5641 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt 5701 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag 5761 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg 5821 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca 5881 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg 5941 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc 6001 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca 6061 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca 6121 gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg 6181 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag 6241 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag 6301 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg 6361 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt 6421 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc 6481 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg 6541 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag 6601 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga 6661 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg 6721 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg 6781 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc 6841 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc 6901 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga 6961 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg 7021 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc 7081 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg 7141 aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc 7201 tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga 7261 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc 7321 tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg 7381 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga 7441 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg 7501 acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc 7561 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg 7621 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg 7681 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc 7741 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca 7801 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg 7861 gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc 7921 tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca 7981 aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg 8041 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg 8101 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca 8161 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct 8221 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg 8281 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg 8341 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg 8401 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg 8461 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc 8521 tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg 8581 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact 8641 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc 8701 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag 8761 gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc tcccccaaga 8821 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt ggtagtggga 8881 gcaacggcag cagcggatcc agcgagctga ttaaggagaa catgcacatg aagctgtaca 8941 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct 9001 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct 9061 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg 9121 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca 9181 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc 9241 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga 9301 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg 9361 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca 9421 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg 9481 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg 9541 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaatggcg 9601 gtggcggagg gatggatgct aagtcactaa ctgcctggtc ccggacactg gtgaccttca 9661 aggatgtatt tgtggacttc accagggagg agtggaagct gctggacact gctcagcaga 9721 tcgtgtacag aaatgtgatg ctggagaact ataagaacct ggtttccttg ggttatcagc 9781 ttactaagcc agatgtgatc ctccggttgg agaagggaga agagcccaaa aggccggcgg 9841 ccacgaaaaa ggccggccag gcaaaaaaga aaaagatcag tctgattgcg gcgttagcgg 9901 tagatcacgt tatcggcatg gaaaccgtca tgccgtggaa cctgcctgcc gatctcgcct 9961 ggtttaaacg caacacctta aataaacccg tgattatggg ccgccatacc tgggaatcaa 10021 tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag cagtcaaccg agtacggacg 10081 atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc ggcgtgtggt gacgtaccag 10141 aaatcatggt tattggcggc ggtcgcgttt atgaacagtt cttgccaaaa gcgcaaaaac 10201 tgtatctgac gcatatcgac gcagaagtgg aaggcgacac ccatttcccg gattacgagc 10261 cggatgactg ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactctcaca 10321 gctattgctt tgagattctg gagcggcgat aaacgcgtga attcactcct caggtgcagg 10381 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag 10441 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt 10501 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 10561 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt 10621 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag 10681 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 10741 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa 10801 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 10861 tagctgtccc tcttctctta tggagatcga tatctcctca tccttccaac ataaatatat 10921 tttgggatta tatcaacatt caatgttact taaagtgacc ttgtaaatat tttcacaact 10981 gagccatgtt tgatttgtat acttatgttt actttactgt ttttcctgaa gttaataatt 11041 gccttgaatt tatttatttc tttaaaaatg tttcattact caggactgta gtttacatta 11101 cgattctttg tgttatacag ttgatgggtt tcttttcttt cttaatttct ttaaaaaata 11161 gagatggggt cttactatat tacccaggct ggtcttgaag tcctgggctc aagtgatctt 11221 cctgtctcag cctaccaagt agctgagact ataggtgcaa aaaagccact atacctggct 11281 agtttacagg ttttaacaaa tgcattatgc cacgtatcca ttattacagg atcacacaag 11341 atattttcat taccctgaaa gcatccctgt gttccaccaa ttcatcctgc ctccatgagc 11401 cgctggcaac cactgatctc tatagttttg ccttttctaa aatgtcatat aattggaatc 11461 atacagtctg tagcattttc agactagctt ttaaaatttg gcaatatgca tttaaggttc 11521 ctccttaaat gtgaagggca tagccaatgt ggcttgatag ctcatttctt tttattggtg 11581 aatatttcat tgtctggatg ttccacagtt tgtttatcca ttcacctatt caatttgctt 11641 tttttctgtg tatctatcac taattcaata ctggactctc caacagagcc gtcccgcttc 11701 cctaaggccc tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc 11761 aggcttccct gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc 11821 cctacagggg ttcctggctc tgctctaagg gcgaattcgc caggaaccgt aaaaaggccg 11881 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 11941 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 12001 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 12061 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 12121 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 12181 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 12241 cagcagccac tggtaaca //