LOCUS Exported 8181 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses RcdCas13a for bacterial expression and contains backbone for spacer cloning. ACCESSION . VERSION . KEYWORDS pC032 - RcdCas13a from Rhodobacter capsulatus DE442 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(1282..1311) /label=lac promoter /note="promoter for the E. coli lac operon" rep_origin 1349..1894 /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 1795..1814 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2011..2028 /label=L4440 /note="L4440 vector, forward primer" promoter 2420..2522 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 2523..3182 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2589..2608) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS complement(4216..5406) /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 5035..5054 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter complement(5556..5590) /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" ORIGIN 1 tttccttatt gctgatgacc tgcctgatgt tcactgctgc agtcaggtcc cggtcgttct 61 tctcagtgtt cacgatcttc cccatagggt cgatccaggc cttcatggat cggcctgcca 121 gggcaaatgc cccgacccac agccgcacga agatttcgct ctccttaatc tcggtctgcc 181 ctgcgctagt ccaataatgg ctctgagcca gatccccggc ctgctgtccc cgatactcgc 241 tgaccctccc catcctgacc atctggttgc gcaccctatt ttcgtgtgtg tgtctcatca 301 gtgccagcag ctctgtcttg tctgctggaa aagctcttgc ggcatttttc cctctggcgc 361 acagtcgctt ataggtcttt ttcacctcgt cgtgcagtgc caggacactt ggcccagctg 421 ctggatcaaa gctgcatctt ttgcttccgg tgaccctttc caggtgctct gccagggcaa 481 acccgacctt tggagacagg ctccgtcctg acacctgtcg aggtgggtca aatgcctgag 541 cagcggacac ggcagccttg gccacttcct tcactgaggc tgccacgtcc acgacgaaat 601 aagtctgaat ttcctcctct ccccagttcc tggccagcct ggccatcccc cttggcagca 661 ctgagctctc gatccccagt gcccttgcga ccaccagccc gggggcaaat ttatcagtct 721 ttgggttcct ctttggctgg ccatcaatcc ttttttcgtc cacatgcaga tgctcataca 781 gtgcagctgc cagttccttt gcgtcgttcc ccctctttga catggcccca taccagcgtc 841 cgtggaaagc ctgtggctca gggggatcgg cctcgaactt cagttttgcg ccgctgtcag 901 cgggctgaaa cttgtcccca taagggtgca gcctgcgctt aaactgatca taatctgagt 961 cctgtgccag ccctgcttca ctcaccagtt tccagaatgc ttcccccaga tcatcccggg 1021 cgtcgaactg cattttgctt ggtgtaggac tgtggatagc cttgccgtct gattttctgc 1081 tatctggttt ccggtagatc ttgtcaatgc cactaatcca ctggccaatc agagcctttg 1141 gatcgcttga caggtgagct ggcagttctt tccggttctt tccgtcagta gaaatcttcc 1201 tcttcagtcc tcctgctggg tccccgaact cgctaatagt ccttccctgc actttgccga 1261 tctgcatgga taagacctcc taacatacga gccggaagca taaagtgtaa atctagaaat 1321 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt 1381 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca 1441 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag 1501 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct 1561 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat 1621 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact 1681 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat 1741 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa 1801 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc 1861 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact 1921 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt 1981 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat 2041 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa 2101 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg 2161 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg 2221 acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca 2281 gtaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc 2341 acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact 2401 tttggcgaaa atgacacgtt gatcggcacg taagaggttc caactttcac cataatgaaa 2461 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta 2521 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag 2581 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg 2641 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta 2701 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg 2761 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg 2821 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat 2881 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg 2941 agaatatgtt tttcgtgtca gccaatccct gggtgagttt caccagtttt gatttaaacg 3001 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag 3061 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc 3121 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt 3181 aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg 3241 ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg 3301 gcagggtcgt taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg 3361 aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg ctctccccgt 3421 ggaggtaata attgacgata tgatcattta ttctgcctcc cagagcctga taaaaacggt 3481 tagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg 3541 cagatccgga acataatggt gcagggcgct tgtttcggcg tgggtatggt ggcaggcccc 3601 gtggccgggg gactgttggg cgctgccggc acctgtccta cgagttgcat gataaagaag 3661 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctaccggca 3721 gcggtgcgga ctgttgtaac tcagaataag aaatgaggcc gctcatggcg ttgactctca 3781 gtcatagtat cgtggtatca ccggttggtt ccactctctg ttgcgggcaa cttcagcagc 3841 acgtagggga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg 3901 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg 3961 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca 4021 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc 4081 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca 4141 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt 4201 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg 4261 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc 4321 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag 4381 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat 4441 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc 4501 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc 4561 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg 4621 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca 4681 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg 4741 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa 4801 ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct tatgcgactc 4861 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa 4921 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc 4981 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat 5041 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg 5101 tccggcgtag aggatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc 5161 tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag 5221 aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact 5281 ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa 5341 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga 5401 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag 5461 cttgttcagt ccgccgtcgt cttggcggtg atgtgaggcc gagacgaatc tccgtctccg 5521 ttcagtccgc cgtcgtcttg gcggtgatgt gaggcgctag cattatacct aggactgagc 5581 tagctgtcaa tcaatcaatc cgcaccaggt cggcagcgtt cagtttggaa gtgttgctct 5641 ttggaggcac gtagatttcc acccggaacc tataccttcg tccgacaatg atagcatcct 5701 cccgaatccg cctcagtcgg gagatgtgca ggccaatatt tccagcgacg ccttcgacag 5761 ccaggaaccc cagagatcca gtatcaatga ctttgacgac cacgccttcg tagacctctc 5821 ccacctgtgg agggggcagc cgggaccctg cggaaggtgc tttgttggga ggtttctgat 5881 ctggctggct ctgggcggtg gctggcttag gaggtttatg ccaagcatct ccgtcgtccc 5941 tgcgccgtgg cttggggttc tgcactgatc cgttaaacac tgcagccacc atctgcaggt 6001 agtcctgaga gaaccgagct tctgtgactg ctgctggtcc gcccacggtc agtctcacct 6061 tgtccaggtg cttgatagct gcctgggtaa tggttgcatc ctgcagcagg tgatccttca 6121 tctgccactt cagggtcagt cccagccgtg ccagcatatc cagaatgctc cttgggacgg 6181 cattttttct cttccgatcg taggccatca tttcccgtgc gcgattcacc agtgcagtca 6241 ggtcgggggt cccatcagcc cgatccagca cgttgaagtg agccagatcc ttccgtgtct 6301 gagttctagc atctgtattg gcggccccgg cgattgtcac tgcccagtct gcgccggccc 6361 ccagctgttt gcttgcattg atcaggaatg tgccagtatc tctttcatag gctccagcat 6421 agtcaatcag gcggccgatc acgtccatca tcagccggtg cagtcgcaga tgatccccca 6481 gatagaccct ccccaccaga aatctatgct cttcgatagt tttgatggct gctgcgtagc 6541 tctggcgctc ctcggggcta attgtcttag ggtggcactt catgaccttg tcgtgcaggt 6601 cggtccgcag ctcctgtgcg gcgacgatct gtgacttctc ctgggtttca tcctcaatct 6661 cggccagcct tgccacctct tcatcgcgca ccttatgttt ggcgaacagg tctgacagga 6721 ctgccatgtg attgtaccgt gcgatctgcc gcagcccccg cagtgtcctt gcgacccgca 6781 gttcaggctc ggatgccccg tccccctctg gatcatcctc ggctggtcga gctgtggtag 6841 gtgttgccat aaacaggcga tcgaatgttg cggggttggc aaacagagct cggaatggct 6901 tcaggtcaaa aggagcagcc cttccctcga agcgtgcctc gccagttttc aggaacagca 6961 ccaggacatc tcggaaccgc ttgaccaggt ccagagcttc ccgccgggcg tcggcctggt 7021 ctgtggcgtc cccgtcctgg accagctcgt acttcccctg cagggcttcc catttccgca 7081 gctggtgcag caggttagac acgtctgagg ctggcacgaa atgcatgacc agatacagtg 7141 cagcctgcca atgctcgtac tgccctcgtg tatcgatagg ttcgggcagc actggtgccc 7201 tagtcatggc tgttgccccg ggctcaattt tcagaatcca atcgaacccc ttggctctga 7261 tataatcctc gaacataaaa gccagcatat ctctccggta attctcaata aactcagcct 7321 gttttcttgc gttttcgctg tcgctttcgt atccgatctg gacccgaaac tctgttgcag 7381 tttccccagt cagtgcagac aggtattctc tcagagtctc tccgtcattg ggaggcctca 7441 gccgtgatga ccgtccttcc atcacgtcag agtaggcctt agtgacattc acggactgag 7501 ccagagctgt tgctgcctct ttggccctag ctgctgggcc agccagtgca gtccctgtaa 7561 tcccggatgc gtaagctctg aatgggccgt catacaggcg cagcagggcg atgtacctgg 7621 ccttggttgc gggatcgtcc agctctcttg gggcaggtgg gggtggcagc tttgttgcga 7681 aggcatgttt tcttgtatcc ctcagcccgt ggacataccc gcgcacccca tcggcccttt 7741 tcagcagcag cttcagcctg ggcagtcctg aactgacttc aggtgcatcc ttcacagcct 7801 tgacgatctc agaatacagt gtagaaaagt gctccttgct tgcgtagtgg gcgacaaagg 7861 cgccggacag atctgccagc agtctggccc ccagtgcttt ggcatcattg tcgatgatag 7921 ctcggatagc tgccacagtt ttgtctgtca ggaccacatg ctctgcctct ttggccttcc 7981 cccaccgggt cttttccagc tctttccgaa tctctttcag aaagccagct cttgccccca 8041 gatgaaatgt ctgatttcta cagcccctca ggtatcgcag cagagcgaac acgaacccct 8101 cgtttccttc tgcccccagt cggtccagct ctggagtctc cccgaagtag attccccttc 8161 tagccattgc ctcggcgacc a //