LOCUS Exported 8181 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses RcsCas13a for bacterial expression and contains backbone for spacer cloning. ACCESSION . VERSION . KEYWORDS pC030 - RcsCas13a from Rhodobacter capsulatus SB 1003 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8181) 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 8181) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8181 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(122..156) /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" promoter complement(4029..4058) /label=lac promoter /note="promoter for the E. coli lac operon" rep_origin 4096..4641 /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 4542..4561 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4758..4775 /label=L4440 /note="L4440 vector, forward primer" promoter 5167..5269 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 5270..5929 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(5336..5355) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS complement(6963..8153) /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 7782..7801 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 aactgtgata aaataccgca ttaaagcttg ttcagtccgc cgtcgtcttg gcggtgatgt 61 gaggccgaga cgaatctccg tctccgttca gtccgccgtc gtcttggcgg tgatgtgagg 121 cgctagcatt atacctagga ctgagctagc tgtcaatcag tcaattcgca ccaggtcagc 181 ggcgttcagt ttggaagtgt tgctcttggg ggggacatag atttcgaccc tgaatctgta 241 tctccgcccg acaatgatgg cgtcctctct gattcgtcgc aggcgggaga tatgcaggcc 301 aatattgcca gccaccccct ccacggccag aaagcccagt gagccggtat caatgacttt 361 gaccacgact ccttcgtaga cctccccgac ctgtgggggt ggcagcctgg accctgcaga 421 aggtgctttg ttgggaggtt tctgatcagg ctgagactga gcagttgcag gcttgggtgg 481 cttgtgccag gcgtccccat catcccggcg ccgtggttta gggttctgca cgctcccatt 541 gaacactgca gcgaccatct gcaggtagtc ctgtgagaag cgtgcttcgg tgacggcggc 601 ggggcccccc acggtcagcc gcactttatc caggtgtttg attgcagcct gggtgattgt 661 agcgtcctgc agcaggtggt ccttcatctg ccacttcagt gtcagcccca gccgagccag 721 catatccagg atactccgtg ggactgcgtt tttgcgtttg cgatcgtagg ccatcatttc 781 tcttgccctg ttcaccagag ctgtcaggtc gggggttccg tcggctctat ccagcacgtt 841 gaaatgtgcc aggtctttcc gggtctgggt gcgtgcgtcg gtattagcgg cgcctgcaat 901 ggtcactgcc caatcagcgc cagcccccag ctgtttgctg gcgttgatca ggaaagtgcc 961 agtatcgcgc tcataagcgc cagcgtaatc aatcaggcgg ccgatgacat ccatcatcag 1021 gcgatgcagc cgcaggtgat cccccaggta cactctgccc accaggaacc tgtgctcctc 1081 aatggttttg attgcggcgg cgtaactctg ccgctcctcg gggctgatgg ttttagggtg 1141 gcatttcatg actttatcgt gcagatctgt cctcagttcc tgtgcggcga caatctgaga 1201 tttttcctga gtttcgtcct caatctcggc caggcgtgcg acttcctcat cccgcacttt 1261 atgtttagcg aacagatcgc tcagcactgc catgtggtta tatcgtgcaa tctggcgcag 1321 cccgcgcagt gtcctggcga cccgcagctc tggttcggag gctccgtcgc cctcggggtc 1381 atcttcagcg ggcctagcag tggttggtgt ggccataaac agccgatcga aagtagctgg 1441 attagcaaac agtgcccgga aaggcttcag atcaaatgga gcggcgcggc cttcaaatct 1501 ggcttcccct gttttcagaa acaggaccag cacatcgcgg aaccttttca ccagatccag 1561 ggcctccctc cgggcatctg cctggtctgt ggcgtcccca tcctggacca gctcgtattt 1621 gccctgcagt gcttcccact ttctcagctg gtgcagcagg ttgctcacat cactagctgg 1681 gacgaagtgc atgaccaggt acagtgcggc ctgccagtgt tcatactggc ctcgtgtatc 1741 gattggctct ggcagcacag gggctctggt cattgctgtg gccccaggtt caatcttcag 1801 aatccagtcg aacccctttg ctcgaatgta atcctcgaac atgaaggcca gcatatctcg 1861 cctatagttc tcgataaact cggcctgctt tctagcattc tctgagtcgg actcatagcc 1921 aatctgcacc cggaactctg tggcagtctc gccagtcagt gcactcaggt attccctcag 1981 ggtttcgcca tcattaggtg gcctcagtcg tgaggtgcgt ccctccatca catcggaata 2041 tgctttagtg acattcacgg actgggccag tgcagtagct gcctctttag ctctggcagc 2101 aggtcctgcc agggcagttc cagtgattcc acttgcgtag gcccgaaatg ggccatcata 2161 cagccgcagc agagcgatat acctggcctt tgttgcaggg tcgtccagtt cccgtggagc 2221 tggaggaggt ggcagcttag tggcgaaagc atgttttcgg gtatcccgca gtccatgcac 2281 gtatcccctg actccgtctg cccgtttcag cagcagcttc agccgaggca gcccggatga 2341 cacttcagga gcgtctttca ctgctttgac gatctcggaa tacagtgtgc tgaagtgttc 2401 tttgcttgca tagtgtgcca caaaagctcc gctcaggtct gccagcagcc gggcgcccag 2461 ggctttggcg tcattgtcaa tgattgctct aatggctgcc actgtcttgt cggtcaggac 2521 cacatgctcg gcttcttttg ccttgcccca ccgtgtcttt tccagctcct tgcggatttc 2581 cttcagaaat cctgcccgag cccccagatg aaaagtctgg tttctacacc ctctcaggta 2641 tcgcagcaga gcgaacacga agccttcatt tccctctgcc cccagtctat ccagctctgg 2701 tgtctcgccg aaatagattc cgcgccgagc cattgcctca gcgaccatct ccttgttgga 2761 gattacctgt ctgatgttga cggcggcagt cagatctcta tcattcttct cagtgttcac 2821 gatcttcccc attgggtcaa tccatgcttt catgctgcgc ccagccagtg caaaagctcc 2881 cacccacagc ctgacaaaga tctcgctttc tttgatttct gtctggccgg cacttgtcca 2941 atagtgggac tgagccagat ccccggcctg ctggcctcgg tattctgaca cccttcccat 3001 tctgaccatc tgattccgga ctctgttctc atgggtatgt ctcatcagag ccagcagttc 3061 ggtcttgtcg gctggaaaag ctctggctgc attctttccc cgggcgcaca gtcttttata 3121 agttttcttc acttcgtcgt gcagtgccag gacagaaggg ccggctgcgg gatcgaatga 3181 gcacctcttg gagcctgtca ccctttccag atgctctgcc agtgcgaacc ccactttggg 3241 actcagactt ctcccgctga cctggcgagg tggatcgaaa gcctgtgctg cgctcacagc 3301 agctttagcc acctccttca ctgatgcggc cacatccacg acaaagtaag tctggatttc 3361 ctcttcgccc caattcctag ccagtctagc cattcctctt ggcagcacgc ttgactcaat 3421 ccccagagcg cgggccacca ccagccctgg agcgaattta tcagtcttag gattccgctt 3481 aggctggcca tcgattcttt tctcatcgac gtgcagatgt tcgtacagag cgactgccag 3541 ctcctttgcg tcattcccgc gcttggacat tgctccatac cacctgccgt ggaaggcctg 3601 aggctcaggt gggtcggctt cgaacttcag ttttgctccg ctatcagcag gctgaaattt 3661 gtctccgtat ggatgcagtc gccgcttaaa ctgatcatag tcactatcct gagccaggcc 3721 agcttctgac accagtttcc agaatgcctc tcccaggtca tcccttgcgt caaactgcat 3781 tttgctaggg gttggggagt ggattgcttt gccatcggat ttcctactat cgggcttccg 3841 gtagattttg tcgatccctg agatccactg tccaatcagg gcctttggat cacttgacag 3901 gtgtgctggc agttccttgc gattctttcc atcggtgcta atctttctct tcagtcctcc 3961 tgcggggtcc ccaaactcgc taatagtcct tccctgcact tttccaatct gcatggataa 4021 gacctcctaa catacgagcc ggaagcataa agtgtaaatc tagaaatatt ttatctgatt 4081 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga 4141 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg 4201 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc 4261 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg 4321 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4381 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac 4441 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac 4501 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt 4561 tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca 4621 ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta 4681 tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca 4741 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca 4801 tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac 4861 accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggcg 4921 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa 4981 acagaagcca ctggagcacc tcaaaaacac catcatacac taaatcagta agttggcagc 5041 atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata 5101 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg 5161 acacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc 5221 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa 5281 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc 5341 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt 5401 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc 5461 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat 5521 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc 5581 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt 5641 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt 5701 cgtgtcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga 5761 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct 5821 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat 5881 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg 5941 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat 6001 gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa 6061 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc 6121 gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt 6181 gacgatatga tcatttattc tgcctcccag agcctgataa aaacggttag cgcttcgtta 6241 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca 6301 taatggtgca gggcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac 6361 tgttgggcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg 6421 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct accggcagcg gtgcggactg 6481 ttgtaactca gaataagaaa tgaggccgct catggcgttg actctcagtc atagtatcgt 6541 ggtatcaccg gttggttcca ctctctgttg cgggcaactt cagcagcacg taggggactt 6601 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt 6661 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg 6721 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat 6781 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat 6841 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa 6901 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc 6961 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg 7021 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa 7081 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat 7141 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg 7201 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc 7261 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc 7321 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc 7381 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc 7441 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag 7501 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg 7561 ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg cattaggaag 7621 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag 7681 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa 7741 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag 7801 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg 7861 atccacagga cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa 7921 gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat 7981 agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg 8041 gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca 8101 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt 8161 gcctgactgc gttagcaatt t //