LOCUS Exported 8477 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses LshCas13a for bacterial expression and contains backbone for spacer cloning. ACCESSION . VERSION . KEYWORDS pC018 - LshCas13a from Leptotrichia shahii SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8477) AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F TITLE RNA targeting with CRISPR-Cas13. JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049. PUBMED 28976959 REFERENCE 2 (bases 1 to 8477) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8477 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(196..230) /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" promoter complement(4415..4444) /label=lac promoter /note="promoter for the E. coli lac operon" rep_origin 4482..5027 /direction=RIGHT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind 4928..4947 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5144..5161 /label=L4440 /note="L4440 vector, forward primer" promoter 5553..5655 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 5656..6315 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(5722..5741) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS complement(7349..62) /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" primer_bind 8168..8187 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc 61 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagcttg 121 ttttagtccc cttcgatatt ggggtggcga gacgaatctc cgtctccgtt ttagtcccct 181 tcgatattgg ggtgggctag cactatacct aggactgagc tagctgtcaa tcacagggta 241 tcgttagtat tctcgatttt ggtcagcagt tcaatgatca gattcttaat gtagtcactg 301 ttgtaggact ccagttccag gacgctcact ttcttgggct tcatcaggcg ctccagaata 361 tcattgttgc cgatcagttt gaatttcttt ttcagctcat cgtagtccag attcacgtcc 421 tttttaaaga cttcgaacac gctggcataa gttgaattgt tgtagcgtgt gctatatgac 481 agcaggttgc tcacccgatc gatctgctca gcaatactgt agtcggcaaa tggatttctc 541 acgatgtaga aatgggagat atagttccga atgctctcat tttcaggctt gttgatctcg 601 gaattttcgc tcaggtcgat tccaaacccg tagcaaatct tctcgaactt cttgtaggac 661 tcttcatcaa agaacttgta gtaggctgtg gtagtgtaaa agccgtcgct tccattgcgc 721 tttgggtatg ccctgctgat cccggtgtta tagcctgaca gcttaatgat gcccagctcc 781 ctcagtccat tcacgatata gtgcatatcc ctttcgaatc ttgccatctg aatagccagc 841 ttccagttga tgtcaatcag gtaggactcg attttgttca gatagttgaa ctcgaccaga 901 tctcgaatct ttttgtactc agacacgcgg ttatagtcct tttcgaagga tttgtagttc 961 ttattctgga tgttctttgc aaagaaatcg tcgttctcct tcagcttttt gatgtacttc 1021 tccttgtatt cctttgagta gccgttcagt ttatcgttca gattcttcag aatagcgtcg 1081 atctcactga ttttgttctt tctgatgttc tttccatcga tattaaacag gaatttagca 1141 tcggccatct tgatgtcgtt gctaatcagg ccgtagatct tgtcaaagtt aggatttccg 1201 atgttcagga acagattctt tttgtagata tacagctcgt tcttccgttc cttgggatag 1261 tagatctcct ggaacttgtt ctcattttca gactccatat cctcaatcag attgtcgatt 1321 tccttcttgt actttttcag aaagtcactg ttgaaaatga tcctgcacag gattttagac 1381 ttgatctcct gatctttgtc cttgatatac tgatccacct ttttcttcag gtccttcttg 1441 ttgatgttag acagctttct ctgttcatcc tgcaggatgt tggatttctt gtcgatctcg 1501 aacttggttt catcgtcgaa gatgacaatc ttctccagtt tcttttccag cacatcacag 1561 ccgttgatat cgtctttaaa ctcggtcaga atgttgttct tgatgtcctc gtagtagttg 1621 ttgaagattt ctttcttagt ctggatcttg aagtcgtcga aatccttttc gatctctttc 1681 atcttctgaa tgaactcttc caggttcaga ttccagttct ctgtgatgca ttcgttccgc 1741 agggtattca gctgcataat ctcatccagg atgtcaatga tattctggta ttctgaggtg 1801 ttcagccaca cgctggtggc gaagaatctg tttcggatct tattaatcac ggcattactg 1861 ttcagcagtg caaagataga aatgatgtac tcgaaatcgt cgttgatgac aatggttttg 1921 tcgctggttt tcacggtgat ccactcataa gtcttgttat cgttgatgtc cttaatctgt 1981 ttcttgattt cctggatgtt catcttgaag tctgaaaagt cgaacagctc ttcgtagttt 2041 ttgcgcagat atccaatgta gcactcgatc actttcttct gatatttctt gattgccttg 2101 ttgttcccct tactggcaga aatctgtgcg ttcttgtagt agttctcgat gatgttctca 2161 tcgatttcgt caatgttgcc cagggttttc ttcagttcct gcaggaagat attcttggac 2221 tcgttctctt ccagatcgtc ctccaggatc agtttcttat acagctcctt attgacgtag 2281 atcagagcgt tcagcacaat tttttcagtc tcgattgtgt caaaaggctc attcttggga 2341 ttgttgcggt acaggttcag gatttcgggc agcactttgc tgaagctggg cagatacttg 2401 atatcgttgt tgttctcttc agagatcttg atgtcattga tctttgtgat gatgtttttc 2461 ttgtccttaa agaccacgtc caggttcaga gctttgctca cctcttcatc ggagatcttc 2521 agattctgaa tgatgttgat cactttgttg taatcgtcct gagtcccctg caggtccctc 2581 tctttggaaa tggcatgcag gatgcggttc ctttcattag tgccgatctt tgtaaacttt 2641 cgaatgaagt tgtttgtgat gttgttcttg ttatcgatga aatccaggtc gcggatgatc 2701 ttgatcttac tattcaggat tttcttatcc agcacatagt tcttctcccg gtcgcctcca 2761 aagaaatcga tgttctcgtc gttgttgatg ttttctcggg aaaagatctt gttcagctcc 2821 atatttgtgc tggcaaagaa ggtgatcagt tccaggtcca gctcttcctt ggcgtgcagg 2881 cggctgaaat cgtcagtatt cactgtggtc atatcgatgt cgttatgcct cagcttcccc 2941 aggtacataa tgtgctccag tgtatactgt ttcactctct tcagaatttt ctctgacagg 3001 atactttcat tcagaatctt ttcgatctcg atcttctcca ttttcttcag tctgaccttc 3061 tgctcgttca ccaggatttt ttcaattctg cccttcaggt atctgtagat gatcttatac 3121 agctccttct cttcatcaga tttcttggaa aatttcttgc tgtcgaagtt caccttgtaa 3181 tgcttcttaa agattccgaa gatctcggta tcacagttcc ctttcttcag ttccttctcc 3241 agtttcttaa tcagctcgtc gatcttgaac tcagccagga tcttctcaat cttttccttg 3301 atagagttgt tcttaatgtt ctcgacaaag aacttcacga tcttatcctt cttgttttct 3361 ctctcgattt taaacttttc gtgcttgtcc agcagcacat aggacttgat gtatgtccga 3421 tttctccgct tctccaggaa ctcgttattg actttcttca ctttctcaat gcgctttgtg 3481 atgttccaaa attccagctc cttgatcacg aaatcggcaa tgtcctcgac ggtcagatcc 3541 acattgatgt tcagaatctt ttccaccagc attttcttat ttttactttt ctttttgtcc 3601 ccgccgacgt tcaggtaaaa cttcacgaag cccaggatct ccagattaga cttgatcttc 3661 tcccggattt ccatgaagtt ggtcaggatc acgtcaatct tatcgtcttt cagcagcttc 3721 tccctcaggt gctcttcgta gtatctattt tcaaacacct tttctgtctc gttttcgatg 3781 atcttctcga tgatcttata caggctcata ttgatgttct tgaagatctc gtagatgctc 3841 ttctttgtct ccagttcatc gttttcaatg atgcgcagaa tgatggagca gtcgttcagg 3901 gtcttattag tgtactcatc ccggatgtca atttcgatct cttcctcgtt ctcctgtcgc 3961 ttgatctcga tctttttatc gtccttagta atgccctgcc tgatggcctc gtcaatgatc 4021 tttttctttg tgatccccag tgctttcagc ttctcgctct tgccgtaggc ctcgatatac 4081 agcaccactt cctctgtctc caggaagtcg tcgttattct caattctaat gatcccttct 4141 ttgcccttca gtttgaacag gatatttcct gcatgaaatt tcctagtgaa ctccttcaga 4201 atattatcgt ttttcttgta gttgatgtac tttctgatga acttgttgtt gtcgatcttc 4261 tctttgttgt tgttttcgtt gatgttcaga atgtacttgt tcccgtcata attccgcttg 4321 actttcacct ttcgcttgat cttgaagtct ttcttatctc gcacctcgta ccaccgcttg 4381 tgtccgaaca ggttccccat ggataagacc tcctaacata cgagccggaa gcataaagtg 4441 taaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg 4501 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt 4561 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa 4621 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca 4681 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg 4741 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag 4801 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc 4861 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc 4921 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg 4981 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg 5041 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg 5101 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga 5161 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct 5221 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata 5281 cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag 5341 ctgaacagga gggacagctg atagaaacag aagccactgg agcacctcaa aaacaccatc 5401 atacactaaa tcagtaagtt ggcagcatca cccgacgcac tttgcgccga ataaatacct 5461 gtgacggaag atcacttcgc agaataaata aatcctggtg tccctgttga taccgggaag 5521 ccctgggcca acttttggcg aaaatgacac gttgatcggc acgtaagagg ttccaacttt 5581 caccataatg aaataagatc actaccgggc gtattttttg agttatcgag attttcagga 5641 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa 5701 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag 5761 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt 5821 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg 5881 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc 5941 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag 6001 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct 6061 aaagggttta ttgagaatat gtttttcgtg tcagccaatc cctgggtgag tttcaccagt 6121 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa 6181 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt 6241 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg 6301 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac 6361 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg 6421 tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt 6481 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc 6541 aggctctccc cgtggaggta ataattgacg atatgatcat ttattctgcc tcccagagcc 6601 tgataaaaac ggttagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca 6661 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gcttgtttcg gcgtgggtat 6721 ggtggcaggc cccgtggccg ggggactgtt gggcgctgcc ggcacctgtc ctacgagttg 6781 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa 6841 ggagctaccg gcagcggtgc ggactgttgt aactcagaat aagaaatgag gccgctcatg 6901 gcgttgactc tcagtcatag tatcgtggta tcaccggttg gttccactct ctgttgcggg 6961 caacttcagc agcacgtagg ggacttccgc gtttccagac tttacgaaac acggaaaccg 7021 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt 7081 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg 7141 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag 7201 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg 7261 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat 7321 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc 7381 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt 7441 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag 7501 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct 7561 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca 7621 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt 7681 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag 7741 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca 7801 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct 7861 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc 7921 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgacgctctc 7981 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg 8041 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc 8101 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt 8161 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc 8221 cggccacgat gcgtccggcg tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt 8281 agtcgatagt ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg 8341 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca 8401 cgccatagtg actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta 8461 ccggcataac caagcct //