LOCUS Exported 9291 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes gRNA for 3' target of human GATAD1 along with Cas9 with 2A GFP. ACCESSION . VERSION . KEYWORDS PX458_GATAD1_2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9291) AUTHORS Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts BS, Mackiewicz M, Mendenhall EM, Myers RM TITLE CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. JOURNAL Genome Res. 2015 Sep 9. PUBMED 26355004 REFERENCE 2 (bases 1 to 9291) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9291 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 15..255 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 15..35 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 186..205 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 285..360 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 457..742 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 744..1021 /label=chicken beta-actin promoter intron 1022..1249 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" regulatory 1261..1270 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1270..1335 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1342..1362 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1387..5487 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5488..5535 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 5551..5604 /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 5605..6318 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 5605..6318 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(5647..5668) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5908..5927) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6255..6276 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(6346..6363) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6352..6559 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6568..6708 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6568..6697 /label=AAV2 ITR rep_origin 6783..7238 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6870..6889) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7080..7101 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7255..7274) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7374..7396 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7434..7452) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7520..7624 /gene="bla" /label=AmpR promoter CDS 7625..8485 /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(7843..7862) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8656..9244 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9145..9164 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca 61 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa 121 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta 181 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata 241 tcttgtggaa aggacgaaac accgtttaat ttgtgaacta caaagtttta gagctagaaa 301 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc 361 ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt 421 gcgccaattc tgcagacaaa tggctctaga ggtacccgtt acataactta cggtaaatgg 481 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg 541 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 601 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 661 ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 721 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat 781 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc 841 gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 901 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 961 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 1021 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 1081 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 1141 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt 1201 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt 1261 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa 1321 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca 1381 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc 1441 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac 1501 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc 1561 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc 1621 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac 1681 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc 1741 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga 1801 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc 1861 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc 1921 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac 1981 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc 2041 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga 2101 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc 2161 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg 2221 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc 2281 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc 2341 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg 2401 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac 2461 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc 2521 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg 2581 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg 2641 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag 2701 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac 2761 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag 2821 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat 2881 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc 2941 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc 3001 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg 3061 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa 3121 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa 3181 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc 3241 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat 3301 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg 3361 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc 3421 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac 3481 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc 3541 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag 3601 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc 3661 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag 3721 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac 3781 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg 3841 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac 3901 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga 3961 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg 4021 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat 4081 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga 4141 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg 4201 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag 4261 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac 4321 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa 4381 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg 4441 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac 4501 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag 4561 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat 4621 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag 4681 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg 4741 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg 4801 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt 4861 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc 4921 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct 4981 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa 5041 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc 5101 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg 5161 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga 5221 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat 5281 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga 5341 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc 5401 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg 5461 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag 5521 gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt 5581 gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg 5641 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 5701 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 5761 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 5821 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 5881 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 5941 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 6001 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 6061 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 6121 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 6181 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 6241 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 6301 atggacgagc tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt 6361 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 6421 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 6481 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga 6541 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct 6601 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 6661 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc 6721 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata 6781 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 6841 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 6901 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 6961 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg 7021 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 7081 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt 7141 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt 7201 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat 7261 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 7321 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 7381 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 7441 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 7501 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 7561 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 7621 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 7681 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 7741 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 7801 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 7861 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 7921 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 7981 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 8041 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 8101 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 8161 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 8221 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 8281 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 8341 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 8401 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 8461 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 8521 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 8581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 8641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 8701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 8761 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 8821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 8881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 8941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 9001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 9061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 9121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 9181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 9241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc c //