LOCUS Exported 9444 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses BE4-Gam in mammalian cells. ACCESSION . VERSION . KEYWORDS BE4-Gam SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9444) AUTHORS Komor AC, Zhao KT, Packer MS, Gaudelli NM, Waterbury AL, Koblan LW, Kim YB, Badran AH, Liu DR TITLE Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity. JOURNAL Sci Adv. 2017 Aug 30;3(8):eaao4774. doi: 10.1126/sciadv.aao4774. eCollection 2017 Aug. PUBMED 28875174 REFERENCE 2 (bases 1 to 9444) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9444 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 132..335 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 286..306 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 377..396 /label=T7 /note="T7 promoter, forward primer" promoter 377..395 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 403..412 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 979..1662 /codon_start=1 /product="cytidine deaminase (C to U editing enzyme) from rat" /label=APOBEC-1 /note="can use ssDNA as a substrate (Komor et al., 2016)" /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL WATGLK" CDS 1759..5859 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5890..6138 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 6169..6417 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 6430..6450 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6459..6476 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6499..6516) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6505..6729 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6800..6816) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6800..6816) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6813..6835) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6824..6840 /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(6848..6878) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6893..6914 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7031..7048) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7202..7790) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7282..7301) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7961..8821) /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 8584..8603 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8822..8926) /gene="bla" /label=AmpR promoter primer_bind complement(9005..9024) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 9196..131 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggctaaacca 421 gcaaaacgta tcaagagtgc cgcagcggct tatgtgccac aaaaccgcga tgcggtgatt 481 accgatatta aacgcatcgg ggatttacag cgcgaagcat cacgtctgga aacggaaatg 541 aatgatgcca tcgcggaaat tacggagaaa tttgcggccc ggattgcacc gattaaaacc 601 gatattgaaa ccctttcaaa aggcgttcag ggatggtgtg aagcgaaccg cgacgaactg 661 acgaacggcg gcaaagtgaa gacggcgaat cttgtcaccg gtgatgtatc gtggcgggtc 721 cgtccaccat cagtaagtat tcgtggtatg gatgcagtga tggaaacgct ggagcgtctt 781 ggcctgcaac gctttattcg cacgaagcag gaaatcaaca aggaagcgat tttactggaa 841 ccgaaagcgg tcgcaggcgt tgccggaatt acagttaaat caggcattga ggatttttct 901 attattccat ttgaacagga agccggtatt agcggcagcg agactcccgg gacctcagag 961 tccgctacac ccgaaagtag ctcagagact ggcccagtgg ctgtggaccc cacattgaga 1021 cggcggatcg agccccatga gtttgaggta ttcttcgatc cgagagagct ccgcaaggag 1081 acctgcctgc tttacgaaat taattggggg ggccggcact ccatttggcg acatacatca 1141 cagaacacta acaagcacgt cgaagtcaac ttcatcgaga agttcacgac agaaagatat 1201 ttctgtccga acacaaggtg cagcattacc tggtttctca gctggagccc atgcggcgaa 1261 tgtagtaggg ccatcactga attcctgtca aggtatcccc acgtcactct gtttatttac 1321 atcgcaaggc tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg ggatttgatc 1381 tcttcaggtg tgactatcca aattatgact gagcaggagt caggatactg ctggagaaac 1441 tttgtgaatt atagcccgag taatgaagcc cactggccta ggtatcccca tctgtgggta 1501 cgactgtacg ttcttgaact gtactgcatc atactgggcc tgcctccttg tctcaacatt 1561 ctgagaagga agcagccaca gctgacattc tttaccatcg ctcttcagtc ttgtcattac 1621 cagcgactgc ccccacacat tctctgggcc accgggttga aatctggtgg ttcttctggt 1681 ggttctagcg gcagcgagac tcccgggacc tcagagtccg ccacacccga aagttctggt 1741 ggttcttctg gtggttctga taaaaagtat tctattggtt tagccatcgg cactaattcc 1801 gttggatggg ctgtcataac cgatgaatac aaagtacctt caaagaaatt taaggtgttg 1861 gggaacacag accgtcattc gattaaaaag aatcttatcg gtgccctcct attcgatagt 1921 ggcgaaacgg cagaggcgac tcgcctgaaa cgaaccgctc ggagaaggta tacacgtcgc 1981 aagaaccgaa tatgttactt acaagaaatt tttagcaatg agatggccaa agttgacgat 2041 tctttctttc accgtttgga agagtccttc cttgtcgaag aggacaagaa acatgaacgg 2101 caccccatct ttggaaacat agtagatgag gtggcatatc atgaaaagta cccaacgatt 2161 tatcacctca gaaaaaagct agttgactca actgataaag cggacctgag gttaatctac 2221 ttggctcttg cccatatgat aaagttccgt gggcactttc tcattgaggg tgatctaaat 2281 ccggacaact cggatgtcga caaactgttc atccagttag tacaaaccta taatcagttg 2341 tttgaagaga accctataaa tgcaagtggc gtggatgcga aggctattct tagcgcccgc 2401 ctctctaaat cccgacggct agaaaacctg atcgcacaat tacccggaga gaagaaaaat 2461 gggttgttcg gtaaccttat agcgctctca ctaggcctga caccaaattt taagtcgaac 2521 ttcgacttag ctgaagatgc caaattgcag cttagtaagg acacgtacga tgacgatctc 2581 gacaatctac tggcacaaat tggagatcag tatgcggact tatttttggc tgccaaaaac 2641 cttagcgatg caatcctcct atctgacata ctgagagtta atactgagat taccaaggcg 2701 ccgttatccg cttcaatgat caaaaggtac gatgaacatc accaagactt gacacttctc 2761 aaggccctag tccgtcagca actgcctgag aaatataagg aaatattctt tgatcagtcg 2821 aaaaacgggt acgcaggtta tattgacggc ggagcgagtc aagaggaatt ctacaagttt 2881 atcaaaccca tattagagaa gatggatggg acggaagagt tgcttgtaaa actcaatcgc 2941 gaagatctac tgcgaaagca gcggactttc gacaacggta gcattccaca tcaaatccac 3001 ttaggcgaat tgcatgctat acttagaagg caggaggatt tttatccgtt cctcaaagac 3061 aatcgtgaaa agattgagaa aatcctaacc tttcgcatac cttactatgt gggacccctg 3121 gcccgaggga actctcggtt cgcatggatg acaagaaagt ccgaagaaac gattactcca 3181 tggaattttg aggaagttgt cgataaaggt gcgtcagctc aatcgttcat cgagaggatg 3241 accaactttg acaagaattt accgaacgaa aaagtattgc ctaagcacag tttactttac 3301 gagtatttca cagtgtacaa tgaactcacg aaagttaagt atgtcactga gggcatgcgt 3361 aaacccgcct ttctaagcgg agaacagaag aaagcaatag tagatctgtt attcaagacc 3421 aaccgcaaag tgacagttaa gcaattgaaa gaggactact ttaagaaaat tgaatgcttc 3481 gattctgtcg agatctccgg ggtagaagat cgatttaatg cgtcacttgg tacgtatcat 3541 gacctcctaa agataattaa agataaggac ttcctggata acgaagagaa tgaagatatc 3601 ttagaagata tagtgttgac tcttaccctc tttgaagatc gggaaatgat tgaggaaaga 3661 ctaaaaacat acgctcacct gttcgacgat aaggttatga aacagttaaa gaggcgtcgc 3721 tatacgggct ggggacgatt gtcgcggaaa cttatcaacg ggataagaga caagcaaagt 3781 ggtaaaacta ttctcgattt tctaaagagc gacggcttcg ccaataggaa ctttatgcag 3841 ctgatccatg atgactcttt aaccttcaaa gaggatatac aaaaggcaca ggtttccgga 3901 caaggggact cattgcacga acatattgcg aatcttgctg gttcgccagc catcaaaaag 3961 ggcatactcc agacagtcaa agtagtggat gagctagtta aggtcatggg acgtcacaaa 4021 ccggaaaaca ttgtaatcga gatggcacgc gaaaatcaaa cgactcagaa ggggcaaaaa 4081 aacagtcgag agcggatgaa gagaatagaa gagggtatta aagaactggg cagccagatc 4141 ttaaaggagc atcctgtgga aaatacccaa ttgcagaacg agaaacttta cctctattac 4201 ctacaaaatg gaagggacat gtatgttgat caggaactgg acataaaccg tttatctgat 4261 tacgacgtcg atcacattgt accccaatcc tttttgaagg acgattcaat cgacaataaa 4321 gtgcttacac gctcggataa gaaccgaggg aaaagtgaca atgttccaag cgaggaagtc 4381 gtaaagaaaa tgaagaacta ttggcggcag ctcctaaatg cgaaactgat aacgcaaaga 4441 aagttcgata acttaactaa agctgagagg ggtggcttgt ctgaacttga caaggccgga 4501 tttattaaac gtcagctcgt ggaaacccgc caaatcacaa agcatgttgc acagatacta 4561 gattcccgaa tgaatacgaa atacgacgag aacgataagc tgattcggga agtcaaagta 4621 atcactttaa agtcaaaatt ggtgtcggac ttcagaaagg attttcaatt ctataaagtt 4681 agggagataa ataactacca ccatgcgcac gacgcttatc ttaatgccgt cgtagggacc 4741 gcactcatta agaaataccc gaagctagaa agtgagtttg tgtatggtga ttacaaagtt 4801 tatgacgtcc gtaagatgat cgcgaaaagc gaacaggaga taggcaaggc tacagccaaa 4861 tacttctttt attctaacat tatgaatttc tttaagacgg aaatcactct ggcaaacgga 4921 gagatacgca aacgaccttt aattgaaacc aatggggaga caggtgaaat cgtatgggat 4981 aagggccggg acttcgcgac ggtgagaaaa gttttgtcca tgccccaagt caacatagta 5041 aagaaaactg aggtgcagac cggagggttt tcaaaggaat cgattcttcc aaaaaggaat 5101 agtgataagc tcatcgctcg taaaaaggac tgggacccga aaaagtacgg tggcttcgat 5161 agccctacag ttgcctattc tgtcctagta gtggcaaaag ttgagaaggg aaaatccaag 5221 aaactgaagt cagtcaaaga attattgggg ataacgatta tggagcgctc gtcttttgaa 5281 aagaacccca tcgacttcct tgaggcgaaa ggttacaagg aagtaaaaaa ggatctcata 5341 attaaactac caaagtatag tctgtttgag ttagaaaatg gccgaaaacg gatgttggct 5401 agcgccggag agcttcaaaa ggggaacgaa ctcgcactac cgtctaaata cgtgaatttc 5461 ctgtatttag cgtcccatta cgagaagttg aaaggttcac ctgaagataa cgaacagaag 5521 caactttttg ttgagcagca caaacattat ctcgacgaaa tcatagagca aatttcggaa 5581 ttcagtaaga gagtcatcct agctgatgcc aatctggaca aagtattaag cgcatacaac 5641 aagcacaggg ataaacccat acgtgagcag gcggaaaata ttatccattt gtttactctt 5701 accaacctcg gcgctccagc cgcattcaag tattttgaca caacgataga tcgcaaacga 5761 tacacttcta ccaaggaggt gctagacgcg acactgattc accaatccat cacgggatta 5821 tatgaaactc ggatagattt gtcacagctt gggggtgact ctggtggttc tggaggatct 5881 ggtggttcta ctaatctgtc agatattatt gaaaaggaga ccggtaagca actggttatc 5941 caggaatcca tcctcatgct cccagaggag gtggaagaag tcattgggaa caagccggaa 6001 agcgatatac tcgtgcacac cgcctacgac gagagcaccg acgagaatgt catgcttctg 6061 actagcgacg cccctgaata caagccttgg gctctggtca tacaggatag caacggtgag 6121 aacaagatta agatgctctc tggtggttct ggaggatctg gtggttctac taatctgtca 6181 gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat cctcatgctc 6241 ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact cgtgcacacc 6301 gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc ccctgaatac 6361 aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa gatgctctct 6421 ggtggttctc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat caccattgag 6481 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 6541 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 6601 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 6661 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 6721 gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct 6781 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga 6901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 7681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8941 aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc 9001 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 9061 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 9121 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg 9181 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt 9241 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 9301 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 9361 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 9421 acttggcagt acatcaagtg tatc //