LOCUS Exported 8181 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses RcrCas13a for bacterial expression and contains backbone for spacer cloning. ACCESSION . VERSION . KEYWORDS pC031 - RcrCas13a from Rhodobacter capsulatus R121 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 12, 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" CDS 59..1249 /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 complement(411..430) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(2283..2942) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 2857..2876 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(2943..3045) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind complement(3437..3454) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3571..4116) /direction=LEFT /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 complement(3651..3670) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 4154..4183 /label=lac promoter /note="promoter for the E. coli lac operon" promoter 8056..8090 /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" ORIGIN 1 caagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat 61 gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggcat 121 aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagcat 181 cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc tatgcgcacc 241 cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg cttcgctact 301 tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga tcctctacgc 361 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 421 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 481 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 541 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 601 gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca acccagtcag 661 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat 721 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt 781 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct 841 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat 901 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg 961 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca 1021 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc 1081 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc 1141 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg 1201 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg 1261 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga 1321 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc 1381 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg 1441 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa 1501 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca 1561 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcccct 1621 acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac cacgatacta 1681 tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca acagtccgca 1741 ccgctgccgg tagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga 1801 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgcccaac agtcccccgg 1861 ccacggggcc tgccaccata cccacgccga aacaagcgcc ctgcaccatt atgttccgga 1921 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc 1981 gctaaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt caattattac 2041 ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact 2101 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc 2161 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta 2221 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa 2281 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg 2341 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg 2401 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg 2461 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgacac gaaaaacata 2521 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc 2581 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac 2641 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc 2701 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg 2761 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta 2821 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa 2881 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc 2941 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat 3001 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtg tcattttcgc 3061 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg 3121 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac 3181 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag 3241 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca 3301 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa 3361 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca 3421 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag 3481 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca 3541 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca 3601 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca 3661 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg 3721 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt 3781 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg 3841 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg 3901 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc 3961 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa 4021 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca 4081 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata 4141 aaatatttct agatttacac tttatgcttc cggctcgtat gttaggaggt cttatccatg 4201 cagattggga aagtccaggg cagaaccatt agcgagtttg gcgaccccgc aggagggctg 4261 aaaaggaaga ttagcaccga cggaaagaac cggaaagaac tgcccgccca tctgtcaagc 4321 gatccaaagg ctctgatcgg ccagtggatt agcggcattg ataagatcta caggaagcct 4381 gattcaagaa agagcgatgg aaaagccatc catagcccaa ctccaagcaa gatgcagttt 4441 gatgccaggg acgacctggg ggaagcattc tggaaactgg tgtcagaggc tgggctggct 4501 caggacagcg attacgatca gtttaaaagg cggctgcatc cctatgggga taagtttcag 4561 cccgccgata gcggcgctaa actgaaattc gaagctgatc ctccagagcc ccaggctttt 4621 cacggccgct ggtatggggc aatgagcaaa agaggaaacg acgctaaaga actggcagca 4681 gcactgtatg aacatctgca tgtcgatgaa aaacgaattg atggccagcc aaaaaggaac 4741 ccaaagacag ataaattcgc accagggctg gtcgtggcca gggcactggg gatcgagagc 4801 tctgtgctgc cacgggggat ggcaagactg gcacggaatt ggggggaaga ggaaatccag 4861 acatacttcg tggtcgacgt ggcagcaagc gtgaaggaag tggcaaaggc agccgtgagc 4921 gccgcacagg ccttcgaccc acctcggcag gtgtccggca ggtcactgag cccaaaggtc 4981 gggtttgccc tggctgaaca tctggagcgg gtcacaggct ccaaaaggtg cagcttcgat 5041 cctgcagctg ggccaagcgt cctggccctg cacgacgaag tcaagaaaac ctataaacgg 5101 ctgtgcgccc ggggaaagaa cgccgcaaga gcctttccag cagacaaaac agaactgctg 5161 gccctgatgc ggcataccca cgagaaccgg gtgaggaatc agatggtcag aatggggcgg 5221 gtgagtgagt atagggggca gcaggcaggg gacctggcac agagccacta ctggacaagc 5281 gcaggacaga ccgaaatcaa ggagtcagag atttttgtcc gcctgtgggt cggggctttc 5341 gcactggccg ggaggtcaat gaaagcctgg attgatccca tgggaaagat cgtcaacact 5401 gaaaagaacg atcgcgacct gaccgctgca gtcaacattc ggcaggtcat cagcaacaag 5461 gagatggtgg ccgaagctat ggcacgacgg gggatctact ttggagaaac cccagagctg 5521 gataggctgg gagcagaagg caatgagggc ttcgtctttg cactgctgag atacctgaga 5581 gggtgcagga atcagacctt tcacctgggg gccagggcag ggttcctgaa agagattaga 5641 aaagaactgg aaaaaacccg gtgggggaag gccaaggagg cagagcacgt ggtcctgacc 5701 gataaaactg tcgctgccat cagagccatc atcgacaacg atgccaaggc actgggggcc 5761 cggctgctgg cagacctgtc aggggctttt gtcgctcact atgccagtaa agaacacttt 5821 agcaccctgt actcagaaat tgtcaaggct gtgaaggacg caccagaggt gtcatccggg 5881 ctgccccggc tgaagctgct gctgaagagg gcagacggag tcagaggcta tgtccatggg 5941 ctgagggata caaggaaaca cgcctttgct actaaactgc caccacctcc agcaccaagg 6001 gaactggatg acccagccac caaagccaga tacatcgcac tgctgagact gtacgatgga 6061 ccattcaggg cttatgcatc cggcattact gggacagctc tggccgggcc agctgcacga 6121 gccaaggagg ccgcaaccgc tctggcacag tccgtgaatg tcacaaaagc atacagcgat 6181 gtgatggagg gcaggtcaag cagactgagg ccacccaacg atggggaaac actgagagaa 6241 tacctgagcg cactgacagg agaaacagca acagaatttc gggtccagat tggctatgaa 6301 agcgactcag agaatgctcg gaagcaggcc gagttcattg agaactatag acgggatatg 6361 ctggctttta tgtttgagga ctacatccgg gctaaggggt tcgactggat tctgaaaatc 6421 gagcccggag caactgcaat gacacgggca cctgtgctgc ccgagccaat tgacacccgg 6481 gggcagtacg aacactggca ggcagcactg tatctggtaa tgcatttcgt gcccgcatca 6541 gatgtcagca acctgctgca tcagctgagg aagtgggagg ccctgcaggg gaaatatgaa 6601 ctggtgcagg atggcgacgc aacagatcag gcagacgcac gacgagaggc cctggatctg 6661 gtcaagcgat ttcgcgatgt cctggtgctg ttcctgaaaa caggagaggc ccggttcgag 6721 ggacgggcag ctccattcga cctgaaaccc ttccgagcac tgttcgcaaa tccagcaact 6781 ttcgatcggc tgtttatggc tacacctacc acagcaaggc cagcagagga tgatcccgag 6841 ggggacggcg catcagaacc agagctgagg gtggcccgga ctctgagagg gctgcggcag 6901 attgccaggt ataatcacat ggcagtcctg agcgatctgt ttgcaaagca caaagtccga 6961 gatgaggaag tcgcaaggct ggcagaaatt gaagatgaga ctcaggagaa atcacagatc 7021 gtcgctgcac aggaactgag aaccgacctg catgacaaag tcatgaagtg tcatcccaaa 7081 accatctctc ctgaagaaag acagtcctac gccgcagcta ttaagaccat cgaagagcat 7141 aggtttctgg tggggagggt ctatctggga gatcatctgc ggctgcacag actgatgatg 7201 gatgtgattg gacggctgat cgattacgct ggagcctacg agagggatac tgggacattt 7261 ctgatcaatg cctcaaagca gctgggggcc ggcgcagact gggcagtgac aattgccggg 7321 gcagcaaaca ctggcgccag gactcagact cggaaggacc tggcccactt taacgtgctg 7381 gatagggccg acgggacacc tgacctgacc gccctggtga atcgagcaag agaaatgatg 7441 gcatacgatc ggaaaagaaa aaacgcagtc ccaagatcaa ttctggatat gctggccagg 7501 ctgggactga ctctgaaatg gcagatgaag gatcacctgc tgcaggatgc aactattacc 7561 caggctgcaa ttaagcatct ggacaaagtg cggctgacag tgggcgggcc tgcagctgtg 7621 actgaggcac gattctccca ggactacctg cagatggtgg ccgcagtgtt taacgggtct 7681 gtccagaatc caaagccaag gaggagggac gacggcgacg catggcataa accacctaag 7741 ccagcaactg cacagtccca gcccgaccag aagccaccca acaaagcacc tagcgcaggg 7801 tcacggctgc ccccaccaca ggtcggggaa gtctatgaag gggtggtggt gaaagtgatc 7861 gataccgggt cactgggatt cctggcagtc gaaggggtcg cagggaatat cggcctgcat 7921 attagtaggc tgaggagaat cagagaagat gctattatcg tgggcaggag atacaggttt 7981 tgggtcgaga tctacgtgcc ccccaagtct aatacctcca aactgaacgc tgctgacctg 8041 gtgcgaattg actgattgac agctagctca gtcctaggta taatgctagc gcctcacatc 8101 accgccaaga cgacggcgga ctgaacggag acggagattc gtctcggcct cacatcaccg 8161 ccaagacgac ggcggactga a //