LOCUS Exported 9885 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SpCas9 fused to DHFR domains on both the N- and C- termini and an internal loop in mammalian cells. ACCESSION . VERSION . KEYWORDS pX330-NLC-DHFR-SpCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9885) AUTHORS Manna D, Maji B, Gangopadhyay SA, Cox KJ, Zhou Q, Law BK, Mazitschek R, Choudhary A TITLE A Singular System with Precise Dosing and Spatiotemporal Control of CRISPR-Cas9. JOURNAL Angew Chem Int Ed Engl. 2019 May 6;58(19):6285-6289. doi: 10.1002/anie.201900788. Epub 2019 Apr 2. PUBMED 30834641 REFERENCE 2 (bases 1 to 9885) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9885 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 268..343 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 440..725 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" intron 915..1142 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" regulatory 1154..1163 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1163..1228 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1235..1255 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1286..1756 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="ISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFLP KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILER" CDS 3878..6376 /codon_start=1 /product="C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" /label=Cas9(C) /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL IHQSITGLYETRIDLSQLGGD" CDS 6377..6424 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 6425..6901 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="ISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHT WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR" primer_bind complement(6926..6943) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6932..7139 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 7148..7288 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 7148..7277 /label=AAV2 ITR rep_origin 7363..7818 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7450..7469) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7660..7681 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7835..7854) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7954..7976 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8014..8032) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8100..8204 /gene="bla" /label=AmpR promoter CDS 8205..9065 /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 complement(8423..8442) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9236..9824 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9725..9744 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac 421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt 661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 841 ggctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa 901 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg 961 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg 1021 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt 1081 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc 1141 aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga 1201 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat 1261 ccacggagtc ccagcagccg gatccatcag tctgattgcg gcgttagcgg tagattacgt 1321 tatcggcatg gaaaacgcca tgccgtggaa cctgcctgcc gatctcgcct ggtttaaacg 1381 caacacctta aataaacccg tgattatggg ccgccatacc tgggaatcaa tcggtcgtcc 1441 gttgccagga cgcaaaaata ttatcctcag cagtcaaccg agtacggacg atcgcgtaac 1501 gtgggtgaag tcggtggatg aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt 1561 gattggcggc ggtcgcgtta ttgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac 1621 gcatatcgac gcagaagtgg aaggcgacac ccatttcccg gattacgagc cggatgactg 1681 ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactctcaca gctattgctt 1741 tgagattctg gagcggcgag gaagcggaag cggatccgac aagaagtaca gcatcggcct 1801 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag 1861 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg 1921 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag 1981 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga 2041 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga 2101 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca 2161 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc 2221 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct 2281 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt 2341 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa 2401 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct 2461 gcccggcgga agtggcagca tcagcctgat cgctgctctg gcagtagact acgtgatagg 2521 aatggagaat gcaatgccct ggaatctgcc cgcagacctg gcttggttta aaagaaacac 2581 gcttaataaa ccagtaatca tgggaagaca tacttgggag agtatagggc ggcccctgcc 2641 cggccggaaa aatattatac tctcaagtca accgagcact gacgatagag tcacctgggt 2701 gaagtccgtt gatgaggcta tagccgcatg tggagatgtg cctgaaatca tggtaatcgg 2761 tggtggaagg gtcatagagc aattcctccc gaaggctcag aaactttatc ttacacatat 2821 agacgccgag gtggagggcg acacacattt ccctgactat gaacccgatg attgggaaag 2881 cgtatttagt gaatttcacg acgccgacgc acagaactct cacagttatt gcttcgagat 2941 tctggagcga cgaggcagtg gaagcgagaa gaagaatggc ctgttcggaa acctgattgc 3001 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa 3061 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg 3121 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag 3181 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa 3241 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct 3301 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat 3361 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat 3421 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg 3481 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct 3541 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat 3601 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc 3661 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga 3721 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc 3781 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga 3841 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga 3901 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca 3961 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt 4021 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga 4081 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct 4141 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt 4201 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag 4261 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct 4321 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac 4381 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca 4441 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt 4501 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat 4561 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg 4621 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa 4681 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta 4741 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc 4801 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa 4861 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg 4921 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc 4981 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga 5041 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta 5101 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt 5161 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca 5221 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa 5281 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc 5341 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat 5401 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat 5461 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt 5521 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg 5581 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa 5641 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt 5701 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct 5761 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga 5821 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct 5881 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg 5941 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga 6001 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa 6061 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc 6121 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag 6181 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc 6241 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct 6301 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc 6361 tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa 6421 aaagatcagt ctgattgcgg cgttagcggt agatcacgtt atcggcatgg aaaccgtcat 6481 gccgtggaac ctgcctgccg atctcgcctg gtttaaacgc aacaccttaa ataaacccgt 6541 gattatgggc cgccatacct gggaatcaat cggtcgtccg ttgccaggac gcaaaaatat 6601 tatcctcagc agtcaaccga gtacggacga tcgcgtaacg tgggtgaagt cggtggatga 6661 agccatcgcg gcgtgtggtg acgtaccaga aatcatggtt attggcggcg gtcgcgttta 6721 tgaacagttc ttgccaaaag cgcaaaaact gtatctgacg catatcgacg cagaagtgga 6781 aggcgacacc catttcccgg attacgagcc ggatgactgg gaatcggtat tcagcgaatt 6841 ccacgatgct gatgcgcaga actctcacag ctattgcttt gagattctgg agcggcgata 6901 agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 6961 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 7021 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 7081 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag 7141 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc 7201 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg 7261 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat 7321 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg 7381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct 7441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 7501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg 7561 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 7621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 7681 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt 7741 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa 7801 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt 7861 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 7921 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 7981 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg 8041 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc 8101 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 8161 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 8221 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 8281 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 8341 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 8401 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 8461 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 8521 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 8581 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 8641 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 8701 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 8761 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 8821 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 8881 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag 8941 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 9001 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 9061 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 9121 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 9181 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 9241 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 9301 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 9361 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 9421 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 9481 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 9541 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 9601 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 9661 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 9721 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 9781 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 9841 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt //