LOCUS Exported 9731 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses dcas9 from Streptococcus thermophilus #3; induced with vanillate; for homologous recombination at the vanillate locus in Caulobacter crescentus; tetracycline resistance. ACCESSION . VERSION . KEYWORDS pVCERC-5 Pvan-dcas9 (Sth3) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9731) AUTHORS Guzzo M, Castro LK, Reisch CR, Guo MS, Laub MT. TITLE A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus JOURNAL mBio REFERENCE 2 (bases 1 to 9731) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9731 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 74..90 /label=SK primer /note="common sequencing primer, one of multiple similar variants" terminator 91..137 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(6247..6263) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6274..6290) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin complement(6488..7076) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6568..6587) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(7182..7198) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(7295..8494) /codon_start=1 /gene="tetA" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR " CDS 8600..9250 /codon_start=1 /gene="tetR" /product="tetracycline resistance regulatory protein" /label=TetR /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD" oriT 9544..9653 /note="incP origin of transfer" ORIGIN 1 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagatc 61 cgtcgacggc cgctctagaa ctagtggatc caaataaaac gaaaggctca gtcgaaagac 121 tgggcctttc gttttatctg ttgtttgtcg cattatacgc aaggcgacaa ggtgctgatg 181 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt 241 aatgaattac aacagttttt atataagctt tacgatagcc agcctccggt cgagcgcgcc 301 attgagcctc cgccagctcc cgctggcgcg atcagtggcg cgctcatcgg gcccggcgta 361 tcaggaccgc gtggtcaact tcatcctgaa gcgcccgatc gcccgaatca tgatcggcgc 421 ggcgtcaacg atcacccctc gacggcttcc gcgcccgcct caaccgtggc cccgcgacgg 481 agcttggcca gcaggcggct gaactgggcc cgttcttcct cggtgagggc ggcgaaggcc 541 tcgcgatcgt gagccagcgt cgcttcctca atgcgggtgc ggaggatctc gccagcggcg 601 gtcaggtgca gggcgttggt gcgacggtcg cggcctggcc gccgctcgat cgcgcccaga 661 gcctccagtt catcgacggc cttcatcgtg ctggcgcggt tgatccccag ggctcgtcca 721 agggtgacct ggtcgcagcc ggcgttcagg gcgatgtaga cgatcgccgt ggcgcgcgcg 781 ggggtcacgt tcagggacgt gagacgggcc gccaatccgg ccatgatccg cacatccgtc 841 gagtgcagtt gcagtcccag ccgctctccc agagcgccga aggcggcggc ctcttccatg 901 gtccacaagc ctcaatagca cgtcaaagaa actgtattgc gatggtacta tagcgcttct 961 tccgggacct ccagactggt tcacacctaa agcgttcagt cggcgcgaat gctccacgcc 1021 gcgcccagcg gcgcgccggc gctcgccgcg gcctcgaaga ccttggcgtt acgaatggcg 1081 gccagggcat ggtcgcgcat gatccgctcg gccccctcgg cgtcgccgca cgagaccgcg 1141 tccagcaccg cctgatgctg gcgatgggcg gccagaaggt gttcgtactc ggccggcaag 1201 tccatcagat ccagggccag cgcaccggcc gccgcgaacg gctcgaaacc attgcgcgcc 1261 agggcgctct cgacagcgcc gttgcccgca gcgctgacca gcgtgtcatg gaaggcctga 1321 ttgtacgcgg cgtagcgatc caggtcctcg ccgttaagcc gcccagccgc gaacagcgct 1381 tcgccctcgg cgatcaaggc gacgaaccgc gcgtgggtct ccgccgtcat tccgcgctcg 1441 gccagacgcc gggccgcaaa gccctccagc acccctcgca cctcgatggc gtcgcggatc 1501 tggtcgctgg acacgcctcg ggcggcataa ccccgcgcgc ccagccgcac cacgagccct 1561 tcctgctcca gactccgcaa ggccgtgcgc acgggcatgc gcgagacgcc cagcgccgcc 1621 gctgtcggga tttcggcgat acgctcgccg ctcttgatct cgcccgaggc gatcatcttg 1681 cgaagagcca tcatcacgcg ctggcccggc tttatgcgcg gcatgtccat ggctggactc 1741 tagccgaccg actgagacgc tcacaagcgt tcaattggat ccaatcttga cgtccgtttg 1801 attacgatca agattggatc caataggtcg ccgaaccacg atgcgaggaa acgcatatgc 1861 ctgcaggcgc cttaattaat atgcatggta ccttaagatc tcgagctcct gactcaggag 1921 gtatcgatat gaccaagccg tactcgatcg gcctggccat cggcaccaac tcggtgggct 1981 gggccgtgac caccgacaac tacaaggtgc cgtcgaagaa gatgaaggtg ctgggcaaca 2041 cctcgaagaa gtacatcaag aagaacctgc tgggcgtgct gctgttcgac tcgggcatca 2101 ccgccgaggg ccgccgcctg aagcgcaccg cccgccgccg ctacacccgc cgccgcaacc 2161 gcatcctgta cctgcaggag atcttctcga ccgagatggc caccctggac gacgccttct 2221 tccagcgcct ggacgactcg ttcctggtgc cggacgacaa gcgcgactcg aagtacccga 2281 tcttcggcaa cctggtggag gagaaggcct accacgacga gttcccgacc atctaccacc 2341 tgcgcaagta cctggccgac tcgaccaaga aggccgacct gcgcctggtg tacctggccc 2401 tggcccacat gatcaagtac cgcggccact tcctgatcga gggcgagttc aactcgaaga 2461 acaacgacat ccagaagaac ttccaggact tcctggacac ctacaacgcc atcttcgagt 2521 cggacctgtc gctggagaac tcgaagcagc tggaggagat cgtgaaggac aagatctcga 2581 agctggagaa gaaggaccgc atcctgaagc tgttcccggg cgagaagaac tcgggcatct 2641 tctcggagtt cctgaagctg atcgtgggca accaggccga cttccgcaag tgcttcaacc 2701 tggacgagaa ggcctcgctg cacttctcga aggagtcgta cgacgaggac ctggagaccc 2761 tgctgggcta catcggcgac gactactcgg acgtgttcct gaaggccaag aagctgtacg 2821 acgccatcct gctgtcgggc ttcctgaccg tgaccgacaa cgagaccgag gccccgctgt 2881 cgtcggccat gatcaagcgc tacaacgagc acaaggagga cctggccctg ctgaaggagt 2941 acatccgcaa catctcgctg aagacctaca acgaggtgtt caaggacgac accaagaacg 3001 gctacgccgg ctacatcgac ggcaagacca accaggagga cttctacgtg tacctgaaga 3061 agctgctggc cgagttcgag ggcgccgact acttcctgga gaagatcgac cgcgaggact 3121 tcctgcgcaa gcagcgcacc ttcgacaacg gctcgatccc gtaccagatc cacctgcagg 3181 agatgcgcgc catcctggac aagcaggcca agttctaccc gttcctggcc aagaacaagg 3241 agcgcatcga gaagatcctg accttccgca tcccgtacta cgtgggcccg ctggcccgcg 3301 gcaactcgga cttcgcctgg tcgatccgca agcgcaacga gaagatcacc ccgtggaact 3361 tcgaggacgt gatcgacaag gagtcgtcgg ccgaggcctt catcaaccgc atgacctcgt 3421 tcgacctgta cctgccggag gagaaggtgc tgccgaagca ctcgctgctg tacgagacct 3481 tcaacgtgta caacgagctg accaaggtgc gcttcatcgc cgagtcgatg cgcgactacc 3541 agttcctgga ctcgaagcag aagaaggaca tcgtgcgcct gtacttcaag gacaagcgca 3601 aggtgaccga caaggacatc atcgagtacc tgcacgccat ctacggctac gacggcatcg 3661 agctgaaggg catcgagaag cagttcaact cgtcgctgtc gacctaccac gacctgctga 3721 acatcatcaa cgacaaggag ttcctggacg actcgtcgaa cgaggccatc atcgaggaga 3781 tcatccacac cctgaccatc ttcgaggacc gcgagatgat caagcagcgc ctgtcgaagt 3841 tcgagaacat cttcgacaag tcggtgctga agaagctgtc gcgccgccac tacaccggct 3901 ggggcaagct gtcggccaag ctgatcaacg gcatccgcga cgagaagtcg ggcaacacca 3961 tcctggacta cctgatcgac gacggcatct cgaaccgcaa cttcatgcag ctgatccacg 4021 acgacgccct gtcgttcaag aagaagatcc agaaggccca gatcatcggc gacgaggaca 4081 agggcaacat caaggaggtg gtgaagtcgc tgccgggctc gccggccatc aagaagggca 4141 tcctgcagtc gatcaagatc gtggacgagc tggtgaaggt gatgggcggc cgcaagccgg 4201 agtcgatcgt ggtggagatg gcccgcgaga accagtacac caaccagggc aagtcgaact 4261 cgcagcagcg cctgaagcgc ctggagaagt cgctgaagga gctgggctcg aagatcctga 4321 aggagaacat cccggccaag ctgtcgaaga tcgacaacaa cgccctgcag aacgaccgcc 4381 tgtacctgta ctacctgcag aacggcaagg acatgtacac cggcgacgac ctggacatcg 4441 accgcctgtc gaactacgac atcgacgcca tcatcccgca ggccttcctg aaggacaact 4501 cgatcgacaa caaggtgctg gtgtcgtcgg cctcgaaccg cggcaagtcg gacgacgtgc 4561 cgtcgctgga ggtggtgaag aagcgcaaga ccttctggta ccagctgctg aagtcgaagc 4621 tgatctcgca gcgcaagttc gacaacctga ccaaggccga gcgcggcggc ctgtcgccgg 4681 aggacaaggc cggcttcatc cagcgccagc tggtggagac ccgccagatc accaagcacg 4741 tggcccgcct gctggacgag aagttcaaca acaagaagga cgagaacaac cgcgccgtgc 4801 gcaccgtgaa gatcatcacc ctgaagtcga ccctggtgtc gcagttccgc aaggacttcg 4861 agctgtacaa ggtgcgcgag atcaacgact tccaccacgc ccacgacgcc tacctgaacg 4921 ccgtggtggc ctcggccctg ctgaagaagt acccgaagct ggagccggag ttcgtgtacg 4981 gcgactaccc gaagtacaac tcgttccgcg agcgcaagtc ggccaccgag aaggtgtact 5041 tctactcgaa catcatgaac atcttcaaga agtcgatctc gctggccgac ggccgcgtga 5101 tcgagcgccc gctgatcgag gtgaacgagg agaccggcga gtcggtgtgg aacaaggagt 5161 cggacctggc caccgtgcgc cgcgtgctgt cgtacccgca ggtgaacgtg gtgaagaagg 5221 tggaggagca gaaccacggc ctggaccgcg gcaagccgaa gggcctgttc aacgccaacc 5281 tgtcgtcgaa gccgaagccg aactcgaacg agaacctggt gggcgccaag gagtacctgg 5341 acccgaagaa gtacggcggc tacgccggca tctcgaactc gttcaccgtg ctggtgaagg 5401 gcaccatcga gaagggcgcc aagaagaaga tcaccaacgt gctggagttc cagggcatct 5461 cgatcctgga ccgcatcaac taccgcaagg acaagctgaa cttcctgctg gagaagggct 5521 acaaggacat cgagctgatc atcgagctgc cgaagtactc gctgttcgag ctgtcggacg 5581 gctcgcgccg catgctggcc tcgatcctgt cgaccaacaa caagcgcggc gagatccaca 5641 agggcaacca gatcttcctg tcgcagaagt tcgtgaagct gctgtaccac gccaagcgca 5701 tctcgaacac catcaacgag aaccaccgca agtacgtgga gaaccacaag aaggagttcg 5761 aggagctgtt ctactacatc ctggagttca acgagaacta cgtgggcgcc aagaagaacg 5821 gcaagctgct gaactcggcc ttccagtcgt ggcagaacca ctcgatcgac gagctgtgct 5881 cgtcgttcat cggcccgacc ggctcggagc gcaagggcct gttcgagctg acctcgcgcg 5941 gctcggccgc cgacttcgag ttcctgggcg tgaagatccc gcgctaccgc gactacaccc 6001 cgtcgtcgct gctgaaggac gccaccctga tccaccagtc ggtgaccggc ctgtacgaga 6061 cccgcatcga cctggccaag ctgggcgagg gctaaatact ctggccattt ttctttaccg 6121 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcaac gcgtgcggcc 6181 gctgattagc taagcagaag gccatcctga cggatgcctt tttcgcttgc tagctgcagc 6241 ccgggggatc cactagttct agagcggcca ttcactggcc gtcgttttac aacgtcgtga 6301 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 6361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 6421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 6481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 6541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 6601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 6661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 6721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 6781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 6841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 6901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 6961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7141 ttttggtcat acgaggccct ttcgtcttca agaattaatt cactggccgt cgttttacaa 7201 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 7261 ttcgccagca gatccgtccc tatcgtttcc acgatcagcg atcggctcgt tgccctgcgc 7321 cgctccaaag cccgcgacgc agcgccggca ggcagagcaa gtagagggca gcgcctgcaa 7381 tccatgccca cccgttccac gttgttatag aagccgcata gatcgccgtg aagaggaggg 7441 gtccgacgat cgaggtcagg ctggtgagcg ccgccagtga gccttgcagc tgcccctgac 7501 gttcctcatc cacctgcctg gacaacattg cttgcagcgc cggcattccg atgccacccg 7561 aagcaagcag gaccatgatc gggaacgcca tccatccccg tgtcgcgaag gcaagcagga 7621 tgtagcctgt gccgtcggca atcattccga gcatgagtgc ccgcctttcg ccgagccggg 7681 cggctacagg gccggtgatc attgcctggg cgagtgaatg cagaatgcca aatgcggcaa 7741 gcgaaatgcc gatcgtggtc gcgtcccagt gaaagcgatc ctcgccgaaa atgacccaaa 7801 gcgcggccgg cacctgtccg acaagttgca tgatgaagaa gaccgccatc agggcggcga 7861 cgacggtcat gccccgggcc caccggaacg aagcgagcgg gttgagagcc tcccggcgta 7921 acggccggcg ttcgcctttg tgcgactccg gcaaaaggaa acagcccgtc aggaaattga 7981 ggccgttcaa ggctgccgcg gcgaagaacg gagcgtgggg ggagaaaccg cccatcagcc 8041 caccgagcac aggtcccgcg accatcccga acccgaaaca ggcgctcatg aagccgaagt 8101 gccgcgcgcg ctcatcgcca tcagtgatat cggcaatata agcgccggct accgccccag 8161 tcgccccggt gatgccggcc acgatccgcc cgatatagag aacccaaagg aaaggcgctg 8221 tcgccatgat ggcgtagtcg acagtggcgc cggccagcga gacgagcaag attggccgcc 8281 gcccgaaacg atccgacagc gcgcccagca caggtgcgca ggcaaattgc accaacgcat 8341 acagcgccag cagaatgcca tagtgggcgg tgacgtcgtt cgagtgaacc agatcgcgca 8401 ggaggcccgg cagcaccggc ataatcaggc cgatgccgac agcgtcgagc gcgacagtgc 8461 tcagaattac gatcaggggt atgttgggtt tcacgtctgg cctccggacc agcctccgct 8521 ggtccgattg aacgcgcgga ttctttatca ctgataagtt ggtggacata ttatgtttat 8581 cagtgataaa gtgtcaagca tgacaaagtt gcagccgaat acagtgatcc gtgccgccct 8641 ggacctgttg aacgaggtcg gcgtagacgg tctgacgaca cgcaaactgg cggaacggtt 8701 gggggttcag cagccggcgc tttactggca cttcaggaac aagcgggcgc tgctcgacgc 8761 actggccgaa gccatgctgg cggagaatca tacgcattcg gtgccgagag ccgacgacga 8821 ctggcgctca tttctgatcg ggaatgcccg cagcttcagg caggcgctgc tcgcctaccg 8881 cgatggcgcg cgcatccatg ccggcacgcg accgggcgca ccgcagatgg aaacggccga 8941 cgcgcagctt cgcttcctct gcgaggcggg tttttcggcc ggggacgccg tcaatgcgct 9001 gatgacaatc agctacttca ctgttggggc cgtgcttgag gagcaggccg gcgacagcga 9061 tgccggcgag cgcggcggca ccgttgaaca ggctccgctc tcgccgctgt tgcgggccgc 9121 gatagacgcc ttcgacgaag ccggtccgga cgcagcgttc gagcagggac tcgcggtgat 9181 tgtcgatgga ttggcgaaaa ggaggctcgt tgtcaggaac gttgaaggac cgagaaaggg 9241 tgacgattga tcaggaccgc tgccggagcg caacccactc actacagcag agccatgtag 9301 acaacatccc ctcccccttt ccaccgcgtc agacgcccgt agcagcccgc tacgggcttt 9361 ttcatgccct gccctagcgt ccaagcctca cggccgcgct cggcctctct ggcggccttc 9421 tggcgctcct gctgcggcgt ccgctcgtat gatgatatat ttttatcttg tgcaatgtaa 9481 catcagagat tttgagacac aacgtggctt tccccccccc cccccaccgc ggtggcggcc 9541 gccccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga 9601 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac 9661 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg 9721 cgaaaaagga t //