LOCUS Exported 12684 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION DHFR-degron controlled expression of KRAB-dCas9-BFP-KRAB from the AAVS1 locus. ACCESSION . VERSION . KEYWORDS pMTL5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12684) 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 12684) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12684 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..804 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" protein_bind 806..839 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 846..871 /label=SA /note="splice acceptor site" CDS 895..948 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 958..1557 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(958..977) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1454..1474 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(1589..1606) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1595..1819 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 1823..1856 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." enhancer 1859..2238 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2240..2517 /label=chicken beta-actin promoter intron 2518..3534 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3458..3480 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3542..3561 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 3599..4072 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILER" CDS 4118..4303 /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 4325..8428 /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 8432..8458 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 8477..8497 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8504..8524 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8564..9259 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" CDS 9305..9490 /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" primer_bind complement(9585..9604) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 9650..9705 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(9704..9723) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" protein_bind 10054..10087 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 10128..10964 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" rep_origin complement(11012..11600) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11092..11111) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11771..12631) /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 12394..12413 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 tgctttctct gaccagcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt 61 ggcctgggtc acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt 121 cttcctccac tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc 181 cggagcactt ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg 241 tctgggtcct ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct 301 gggttccctt ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct 361 tctccgacgg atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc 421 gtttttctgg acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc 481 ttgctttctt tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt 541 catttgggca gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc 601 ctgtcatggc atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc 661 ccctatgtcc acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct 721 gggaccacct tatattccca gggccggtta atgtggctct ggttctgggt acttttatct 781 gtcccctcca ccccacagtg gggcaataac ttcgtatagc atacattata cgaagttata 841 agcttctgac ctcttctctt cctcccacag ggcctcgaga gatctggcag cggagagggc 901 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctag gctcgagatg 961 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 1021 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 1081 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 1141 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 1201 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 1261 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 1321 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 1381 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 1441 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 1501 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatct 1561 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 1621 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 1681 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 1741 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 1801 gatgcggtgg gctctatggg tcataacttc gtatagcata cattatacga agttattcga 1861 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 1921 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 1981 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 2041 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 2101 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 2161 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 2221 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc 2281 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg 2341 ggggcggggg gggggggggg gcgcgcgccr ggsggggcgg ggsggggcga ggggcggggc 2401 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct 2461 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga 2521 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 2581 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg 2641 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc 2701 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg 2761 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc 2821 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc 2881 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc 2941 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc 3001 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc 3061 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 3121 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc 3181 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 3241 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc 3301 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 3361 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 3421 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 3481 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 3541 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaatgtcg acgccaccat 3601 gatcagtctg attgcggcgt tagcggtaga ttacgttatc ggcatggaaa acgccatgcc 3661 gtggaacctg cctgccgatc tcgcctggtt taaacgcaac accttaaata aacccgtgat 3721 tatgggccgc catacctggg aatcaatcgg tcgtccgttg ccaggacgca aaaatattat 3781 cctcagcagt caaccgagta cggacgatcg cgtaacgtgg gtgaagtcgg tggatgaagc 3841 catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggtc gcgttattga 3901 acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgacgcag aagtggaagg 3961 cgacacccat ttcccggatt acgagccgga tgactgggaa tcggtattca gcgaattcca 4021 cgatgctgat gcgcagaact ctcacagcta ttgctttgag attctggagc ggggcagcgg 4081 atccggcatg gatgctaagt cactaactgc ctggtcccgg acactggtga ccttcaagga 4141 tgtatttgtg gacttcacca gggaggagtg gaagctgctg gacactgctc agcagatcgt 4201 gtacagaaat gtgatgctgg agaactataa gaacctggtt tccttgggtt atcagcttac 4261 taagccagat gtgatcctcc ggttggagaa gggagaagag cccggcggtt ccggcggagg 4321 gtcgatggac aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc 4381 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga 4441 ccggcacagc atcaagaaga acctgatcgg cgccctgctg ttcgacagcg gagaaacagc 4501 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat 4561 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca 4621 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt 4681 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag 4741 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc 4801 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag 4861 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa 4921 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag 4981 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg 5041 caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc 5101 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct 5161 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc 5221 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc 5281 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt 5341 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta 5401 cgccggctac atcgatggcg gagccagcca ggaagagttc tacaagttca tcaagcccat 5461 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct 5521 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct 5581 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa 5641 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa 5701 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga 5761 ggaagtggtg gacaagggcg ccagcgccca gagcttcatc gagcggatga ccaacttcga 5821 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac 5881 cgtgtacaac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt 5941 cctgagcggc gagcagaaaa aagccatcgt ggacctgctg ttcaagacca accggaaagt 6001 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga 6061 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa 6121 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat 6181 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta 6241 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg 6301 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat 6361 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga 6421 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag 6481 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca 6541 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat 6601 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga 6661 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca 6721 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg 6781 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga 6841 cgctatcgtg cctcagagct ttctgaagga cgactccatc gataacaaag tgctgactcg 6901 gagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat 6961 gaagaactac tggcgccagc tgctgaatgc caagctgatt acccagagga agttcgacaa 7021 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag 7081 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat 7141 gaacactaag tacgacgaga acgacaaact gatccgggaa gtgaaagtga tcaccctgaa 7201 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa 7261 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa 7321 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg 7381 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta 7441 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa 7501 gcggcctctg atcgagacaa acggcgaaac aggcgagatc gtgtgggata agggccggga 7561 ctttgccacc gtgcggaaag tgctgtctat gccccaagtg aatatcgtga aaaagaccga 7621 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgacaagct 7681 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt 7741 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag 7801 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat 7861 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc 7921 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga 7981 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc 8041 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt 8101 ggaacagcac aaacactacc tggacgagat catcgagcag atcagcgagt tctccaagag 8161 agtgatcctg gccgacgcta atctggacaa ggtgctgagc gcctacaaca agcacagaga 8221 caagcctatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg 8281 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac 8341 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg 8401 gatcgacctg tctcagctgg gaggcgacgc ctatccctat gacgtgcccg attatgccag 8461 cctgggcagc ggctccccca agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa 8521 agtggacggc attggtagtg ggagcaacgg cagcagcgga tccagcgagc tgattaagga 8581 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac 8641 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga 8701 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa 8761 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg 8821 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga 8881 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac 8941 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct 9001 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg 9061 gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct 9121 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa 9181 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa 9241 actggggcac aagcttaatg gcggtggcgg agggatggat gctaagtcac taactgcctg 9301 gtcccggaca ctggtgacct tcaaggatgt atttgtggac ttcaccaggg aggagtggaa 9361 gctgctggac actgctcagc agatcgtgta cagaaatgtg atgctggaga actataagaa 9421 cctggtttcc ttgggttatc agcttactaa gccagatgtg atcctccggt tggagaaggg 9481 agaagagccc tagtaacgcg tgaattcact cctcaggtgc aggctgccta tcagaaggtg 9541 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc 9601 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa 9661 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat 9721 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat 9781 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa 9841 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt 9901 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt 9961 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc 10021 ttatggagat ccctcgacct gcagcccaag cttataactt cgtatagcat acattatacg 10081 aagttatgcg gccgcaccgg tatgcattta attaagatat cagctttact agggacagga 10141 ttggtgacag aaaagcccca tccttaggcc tcctccttcc tagtctcctg atattgggtc 10201 taacccccac ctcctgttag gcagattcct tatctggtga cacaccccca tttcctggag 10261 ccatctctct ccttgccaga acctctaagg tttgcttacg atggagccag agaggatcct 10321 gggagggaga gcttggcagg gggtgggagg gaaggggggg atgcgtgacc tgcccggttc 10381 tcagtggcca ccctgcgcta ccctctccca gaacctgagc tgctctgacg cggctgtctg 10441 gtgcgtttca ctgatcctgg tgctgcagct tccttacact tcccaagagg agaagcagtt 10501 tggaaaaaca aaatcagaat aagttggtcc tgagttctaa ctttggctct tcacctttct 10561 agtccccaat ttatattgtt cctccgtgcg tcagttttac ctgtgagata aggccagtag 10621 ccagccccgt cctggcaggg ctgtggtgag gaggggggtg tccgtgtgga aaactccctt 10681 tgtgagaatg gtgcgtccta ggtgttcacc aggtcgtggc cgcctctact ccctttctct 10741 ttctccatcc ttctttcctt aaagagtccc cagtgctatc tgggacatat tcctccgccc 10801 agagcagggt cccgcttccc taaggccctg ctctgggctt ctgggtttga gtccttggca 10861 agcccaggag aggcgctcag gcttccctgt cccccttcct cgtccaccat ctcatgcccc 10921 tggctctcct gccccttccc tacaggggtt cctggctctg ctctaagggc gaattcgcca 10981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 11041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 11101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 11161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 11221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 11281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 11341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 11401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 11461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 11521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 11581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 11641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 11701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 11761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 11821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 11881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 11941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 12001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 12061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 12121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 12181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 12241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 12301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 12361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 12421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 12481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 12541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 12601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 12661 tttatcaggg ttgaattcgc cctt //