LOCUS Exported 9464 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses dead (908A) AsCpf1. ACCESSION . VERSION . KEYWORDS WN10150 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9464) AUTHORS Toth E, Czene BC, Kulcsar PI, Krausz SL, Talas A, Nyeste A, Varga E, Huszar K, Weinhardt N, Ligeti Z, Borsy AE, Fodor E, Welker E TITLE Mb- and FnCpf1 nucleases are active in mammalian cells: activities and PAM preferences of four wild-type Cpf1 nucleases and of their altered PAM specificity variants. JOURNAL Nucleic Acids Res. 2018 Sep 20. pii: 5103950. doi: 10.1093/nar/gky815. PUBMED 30239882 REFERENCE 2 (bases 1 to 9464) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9464 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 45..64 /label=T7 /note="T7 promoter, forward primer" promoter 45..63 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 111..4031 /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQR SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES KDLKLQNGISNQDWLAYIQELRN" regulatory 3751..3760 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4032..4079 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 4086..4166 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" primer_bind complement(4240..4257) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4246..4470 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4516..4944 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4603..4622) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4813..4834 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4953..4973) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4958..5287 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5138..5273 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5200..5219 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5354..6148 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5408..5427) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6018..6037 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6322..6443 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6359..6378) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6413..6432 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6492..6508) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6492..6508) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6505..6527) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6516..6532 /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(6540..6570) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6585..6606 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6723..6740) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6894..7479) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6974..6993) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7650..8510) /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 8273..8292 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8511..8615) /gene="bla" /label=AmpR promoter primer_bind complement(8690..8709) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 8881..9260 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 9261..9464 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 9415..9435 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat 61 agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgccacc atgacacagt 121 tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag ctgatcccac 181 agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac aaggcccgca 241 atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc tatgccgacc 301 agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc gactcctata 361 gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc acatatcgca 421 atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc atcaataaga 481 gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc aaggtgctga 541 agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg agcttcgaca 601 agtttacaac ctacttctcc ggcttttatg agaacaggaa gaacgtgttc agcgccgagg 661 atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag tttaaggaga 721 attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag cactttgaga 781 acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg ttttccttcc 841 ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag ctgctgggag 901 gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg ctgaatctgg 961 ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac agattcatcc 1021 ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg gaggagttta 1081 agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg agaaacgaga 1141 acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac ctgacacaca 1201 tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac cactgggata 1261 cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag atcaccaagt 1321 ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg caggagatca 1381 tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc gagatcctgt 1441 cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag caggaggaga 1501 aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg ctggactggt 1561 ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg accggcatca 1621 agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat gccaccaaga 1681 agccctactc cgtggagaag ttcaagctga actttcagat gcctacactg gcctctggct 1741 gggacgtgaa taaggagaag aacaatggcg ccatcctgtt tgtgaagaac ggcctgtact 1801 atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc gagcccacag 1861 agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat gccgccaaga 1921 tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag acccacacaa 1981 cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag gagatctacg 2041 acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc aagaaaaccg 2101 gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca agggattttc 2161 tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca tcctctcagt 2221 ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac atcagcttcc 2281 agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg tacctgttcc 2341 agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg cacacactgt 2401 attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag ctgaatggcc 2461 aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac cggctgggag 2521 agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac accctgtacc 2581 aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat gaggccaggg 2641 ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag gataggcgct 2701 ttaccagcga caagttcttt ttccacgtgc ctatcacact gaactatcag gccgccaatt 2761 ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc gagacaccta 2821 tcatcggcat cgcccggggc gagagaaacc tgatctatat cacagtgatc gactccaccg 2881 gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac cagaagaagc 2941 tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg gtgggcacaa 3001 tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg gacctgatga 3061 tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag agcaagagga 3121 ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc gataagctga 3181 attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg aacccatacc 3241 agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc ttcctgtttt 3301 acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg gaccccttcg 3361 tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc ttcgactttc 3421 tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac agaaatctgt 3481 ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc gagaagaacg 3541 agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc gtgccagtga 3601 tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac gagctgatcg 3661 ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg ccaaagctgc 3721 tggagaatga cgattctcac gccatcgaca ccatggtggc cctgatccgc agcgtgctgc 3781 agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc gtgcgcgatc 3841 tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggcccatg gacgccgatg 3901 ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac ctgaaggaga 3961 gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc tacatccagg 4021 agctgcgcaa caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg 4081 gatcctaccc atacgatgtt ccagattacg cttatcccta cgacgtgcct gattatgcat 4141 acccatatga tgtccccgac tatgcctaag aattctgcag atatccagca cagtggcggc 4201 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt 4261 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc 4321 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt 4381 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca 4441 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct 4501 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 4561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 4621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 4681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 4741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 4801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 4861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 4921 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 4981 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 5041 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 5101 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 5161 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 5221 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 5281 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 5341 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 5401 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 5461 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 5521 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 5581 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 5641 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 5701 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 5761 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 5821 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 5881 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 5941 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 6001 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 6061 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 6121 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 6181 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 6241 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 6301 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 6361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6421 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 6481 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 6541 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 6601 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6661 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6721 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6781 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6841 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6901 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6961 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7021 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7081 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7141 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7201 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7261 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7321 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7381 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt 7441 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 7501 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 7561 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 7621 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 7681 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 7741 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 7801 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 7861 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 7921 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 7981 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 8041 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 8101 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 8161 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 8221 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 8281 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 8341 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 8401 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 8461 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 8521 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 8581 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 8641 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct 8701 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt 8761 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga 8821 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt 8881 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 8941 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 9001 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 9061 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 9121 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 9181 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 9241 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 9301 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 9361 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 9421 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agct //