LOCUS Exported 10463 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses dcas9 (Streptococcus thermophilus #3); repressed with 0.2% glucose, induced with 0.3% xylose; for homologous recombination at the xyl locus in Caulobacter crescentus; tetracycline resistance. ACCESSION . VERSION . KEYWORDS pXGFPC-5 Pxyl-dcas9 (Sth3) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10463) 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 10463) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10463 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 89..105 /label=SK primer /note="common sequencing primer, one of multiple similar variants" terminator 106..152 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(6994..7010) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7021..7037) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin complement(7235..7823) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7315..7334) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(7929..7945) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(8042..9241) /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 9347..9997 /codon_start=1 /gene="tetR" /product="tetracycline resistance regulatory protein" /label=TetR /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD" oriT 10291..10400 /note="incP origin of transfer" ORIGIN 1 cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat 61 gcagcggaaa agatccgtcg acggccgctc tagaactagt ggatccaaat aaaacgaaag 121 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcgcatta tacgcaaggc 181 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat 241 gtcggcagaa tgcttaatga attacaacag tttttatata agcttagcgc taccggactc 301 agatccagat cgaaatgtac cggctgaact ggtggaagca cgttgagcct cgtcaaggac 361 tcatgaagcg cgtccctgct ctcagtcgcc tgctcgttac gcctgaggtg tcaaaacacc 421 gtctgttcat ttggctagac gctagagttc ttccggatca caagctccag gtcgttacac 481 tcgatgacga ctgctcattc ggagttttac actcaaggtt ccacgaggtg tgggctcttg 541 cagcggggtc gtggcacggc tcaggcaatg atccgcgcta caccatctcc accaccttcg 601 agaccttccc cttcccggaa ggcctgacgc ccaacatagc cgccgtcgac tatgagggcg 661 acccgcgcgc ccaggccatc gccgcggcgg cggccgagct gaaccggctg cgggaagcct 721 ggctcaatcc gcccgacctg gtgcggatcg agcccgaggt cgtgcccggc tatcccgacc 781 gcgtgctgcc cgtcagcccc gaggcgggcg ccgagctgaa gaagcgcacc ctgaccaacc 841 tctacaacca gcgcccggcc tggctggaca tggcccacca gcgcctggac gccgccgtcg 901 ccgccgccta cggctggccc gacggcctga ccgacgacga gatcctggag cgcctgttcg 961 ccctcaacca ggagcgcgcg gcggcggggc ggtaggttag acccctccaa aaccccgacc 1021 tccccggcga aggccggggt ccagttacat ccggaacgct tggagtgggc ccgcctacgg 1081 cctcgaccaa cccgcagtcg gcaaggggtt cgatctggac cccggcattc gccggggagg 1141 tcggcagaga ggtcagaacc gcgtcttgca gccctgcttc agatgaaccc cgcgcaggaa 1201 gccgcggcgg gccaggccgg cgtcgacggc ggcgttcatc tggcggtcgg cgggggcagc 1261 gccggtgact tcgcgcacca ggccccggaa cgggatgatg ctgttgatca gggtcttgcc 1321 accggcctcg gccagcttgc cggctcggtt ttcggcgtac tcggcgccga ccttgtagtc 1381 gggccccagc gccgcgttca gcgccttcac ctcattggcc agggcctggc aggacttcag 1441 acccgcgagg ctgtaggggt cgtcaaaggc cttggccagg accggcggca cctcgcgctt 1501 ggacaggccg acgtcgcgca ccggctgggt gccgatgtcg acggccttct tgcccgcctc 1561 gccccagcgg ccggacggtt cctcgggctt ggccggcggc ttgacctggg cgtgggcgct 1621 ggaggccagc agcacggcgg cggctccggc catgatcagc agcttgttcg gcatcaaaag 1681 cgccctcccg acaaaacacg ccacgggaac ggcaagcctc gccgccggtt cccgcccgca 1741 agtccctcgg cacgaactct aaggcctgcg cacgatccat cgccaccaca aagcgtgtca 1801 tcccggccga agcgcagcgg agagccggga cccagggcca caagcacggc gcttcgtcac 1861 cccctgggtc ccggatagcg ctccgcgctt ccgggatgac acgggtttaa gcacttgggt 1921 gagaagccct agttcggcgc gccgcccgct tccaggtcga acgcggcgtc gaaggcgtcg 1981 tcgccaggaa ccgtcccttc cagggcggcg tcgaacaggt cttcgagcga tgaccagatg 2041 aaggccatct gctccatgat cgtcgggacg ctggccggat cgaagccggg gtagaggtct 2101 tccaccagcc acaggcccgt gccgggatcg aaggcgaagc ggcagccgat caggcggaac 2161 tgggcgcggg ccatggtctc gaacagggcc gtcaggtcgc gggcggcgtc caggtcgtcg 2221 tggtccagca ccaggcagcg gatctgcagc cagccgtggt cgggcagcag gtagaaggcg 2281 ccctcgtcct gatcctcgcc cgaaacctcc agcccccggt cgatggcttc gacgacatag 2341 ccggccgcgc ggcaggtgtc ggtgagcgcg gccagcaggg cggcttcctg gtcaggggtc 2401 aggtcggtca tgggcaagag gtccaggtcg tggtttgtcg gcggcttcta gcatggaccg 2461 cccgcgcccg tgaggccgag gatttcgcgc tggtcagaca acctacttgc cgtccccaca 2521 tgttagcgct accaagtgcc gacgaacgcg cgccgccgac ggtgtcggcg cttcagacgc 2581 tcgagttttg gggagacgac catatgcctg caggcgcctt aattaatatg catggtacct 2641 taagatctcg agctcctgac tcaggaggta tcgatatgac caagccgtac tcgatcggcc 2701 tggccatcgg caccaactcg gtgggctggg ccgtgaccac cgacaactac aaggtgccgt 2761 cgaagaagat gaaggtgctg ggcaacacct cgaagaagta catcaagaag aacctgctgg 2821 gcgtgctgct gttcgactcg ggcatcaccg ccgagggccg ccgcctgaag cgcaccgccc 2881 gccgccgcta cacccgccgc cgcaaccgca tcctgtacct gcaggagatc ttctcgaccg 2941 agatggccac cctggacgac gccttcttcc agcgcctgga cgactcgttc ctggtgccgg 3001 acgacaagcg cgactcgaag tacccgatct tcggcaacct ggtggaggag aaggcctacc 3061 acgacgagtt cccgaccatc taccacctgc gcaagtacct ggccgactcg accaagaagg 3121 ccgacctgcg cctggtgtac ctggccctgg cccacatgat caagtaccgc ggccacttcc 3181 tgatcgaggg cgagttcaac tcgaagaaca acgacatcca gaagaacttc caggacttcc 3241 tggacaccta caacgccatc ttcgagtcgg acctgtcgct ggagaactcg aagcagctgg 3301 aggagatcgt gaaggacaag atctcgaagc tggagaagaa ggaccgcatc ctgaagctgt 3361 tcccgggcga gaagaactcg ggcatcttct cggagttcct gaagctgatc gtgggcaacc 3421 aggccgactt ccgcaagtgc ttcaacctgg acgagaaggc ctcgctgcac ttctcgaagg 3481 agtcgtacga cgaggacctg gagaccctgc tgggctacat cggcgacgac tactcggacg 3541 tgttcctgaa ggccaagaag ctgtacgacg ccatcctgct gtcgggcttc ctgaccgtga 3601 ccgacaacga gaccgaggcc ccgctgtcgt cggccatgat caagcgctac aacgagcaca 3661 aggaggacct ggccctgctg aaggagtaca tccgcaacat ctcgctgaag acctacaacg 3721 aggtgttcaa ggacgacacc aagaacggct acgccggcta catcgacggc aagaccaacc 3781 aggaggactt ctacgtgtac ctgaagaagc tgctggccga gttcgagggc gccgactact 3841 tcctggagaa gatcgaccgc gaggacttcc tgcgcaagca gcgcaccttc gacaacggct 3901 cgatcccgta ccagatccac ctgcaggaga tgcgcgccat cctggacaag caggccaagt 3961 tctacccgtt cctggccaag aacaaggagc gcatcgagaa gatcctgacc ttccgcatcc 4021 cgtactacgt gggcccgctg gcccgcggca actcggactt cgcctggtcg atccgcaagc 4081 gcaacgagaa gatcaccccg tggaacttcg aggacgtgat cgacaaggag tcgtcggccg 4141 aggccttcat caaccgcatg acctcgttcg acctgtacct gccggaggag aaggtgctgc 4201 cgaagcactc gctgctgtac gagaccttca acgtgtacaa cgagctgacc aaggtgcgct 4261 tcatcgccga gtcgatgcgc gactaccagt tcctggactc gaagcagaag aaggacatcg 4321 tgcgcctgta cttcaaggac aagcgcaagg tgaccgacaa ggacatcatc gagtacctgc 4381 acgccatcta cggctacgac ggcatcgagc tgaagggcat cgagaagcag ttcaactcgt 4441 cgctgtcgac ctaccacgac ctgctgaaca tcatcaacga caaggagttc ctggacgact 4501 cgtcgaacga ggccatcatc gaggagatca tccacaccct gaccatcttc gaggaccgcg 4561 agatgatcaa gcagcgcctg tcgaagttcg agaacatctt cgacaagtcg gtgctgaaga 4621 agctgtcgcg ccgccactac accggctggg gcaagctgtc ggccaagctg atcaacggca 4681 tccgcgacga gaagtcgggc aacaccatcc tggactacct gatcgacgac ggcatctcga 4741 accgcaactt catgcagctg atccacgacg acgccctgtc gttcaagaag aagatccaga 4801 aggcccagat catcggcgac gaggacaagg gcaacatcaa ggaggtggtg aagtcgctgc 4861 cgggctcgcc ggccatcaag aagggcatcc tgcagtcgat caagatcgtg gacgagctgg 4921 tgaaggtgat gggcggccgc aagccggagt cgatcgtggt ggagatggcc cgcgagaacc 4981 agtacaccaa ccagggcaag tcgaactcgc agcagcgcct gaagcgcctg gagaagtcgc 5041 tgaaggagct gggctcgaag atcctgaagg agaacatccc ggccaagctg tcgaagatcg 5101 acaacaacgc cctgcagaac gaccgcctgt acctgtacta cctgcagaac ggcaaggaca 5161 tgtacaccgg cgacgacctg gacatcgacc gcctgtcgaa ctacgacatc gacgccatca 5221 tcccgcaggc cttcctgaag gacaactcga tcgacaacaa ggtgctggtg tcgtcggcct 5281 cgaaccgcgg caagtcggac gacgtgccgt cgctggaggt ggtgaagaag cgcaagacct 5341 tctggtacca gctgctgaag tcgaagctga tctcgcagcg caagttcgac aacctgacca 5401 aggccgagcg cggcggcctg tcgccggagg acaaggccgg cttcatccag cgccagctgg 5461 tggagacccg ccagatcacc aagcacgtgg cccgcctgct ggacgagaag ttcaacaaca 5521 agaaggacga gaacaaccgc gccgtgcgca ccgtgaagat catcaccctg aagtcgaccc 5581 tggtgtcgca gttccgcaag gacttcgagc tgtacaaggt gcgcgagatc aacgacttcc 5641 accacgccca cgacgcctac ctgaacgccg tggtggcctc ggccctgctg aagaagtacc 5701 cgaagctgga gccggagttc gtgtacggcg actacccgaa gtacaactcg ttccgcgagc 5761 gcaagtcggc caccgagaag gtgtacttct actcgaacat catgaacatc ttcaagaagt 5821 cgatctcgct ggccgacggc cgcgtgatcg agcgcccgct gatcgaggtg aacgaggaga 5881 ccggcgagtc ggtgtggaac aaggagtcgg acctggccac cgtgcgccgc gtgctgtcgt 5941 acccgcaggt gaacgtggtg aagaaggtgg aggagcagaa ccacggcctg gaccgcggca 6001 agccgaaggg cctgttcaac gccaacctgt cgtcgaagcc gaagccgaac tcgaacgaga 6061 acctggtggg cgccaaggag tacctggacc cgaagaagta cggcggctac gccggcatct 6121 cgaactcgtt caccgtgctg gtgaagggca ccatcgagaa gggcgccaag aagaagatca 6181 ccaacgtgct ggagttccag ggcatctcga tcctggaccg catcaactac cgcaaggaca 6241 agctgaactt cctgctggag aagggctaca aggacatcga gctgatcatc gagctgccga 6301 agtactcgct gttcgagctg tcggacggct cgcgccgcat gctggcctcg atcctgtcga 6361 ccaacaacaa gcgcggcgag atccacaagg gcaaccagat cttcctgtcg cagaagttcg 6421 tgaagctgct gtaccacgcc aagcgcatct cgaacaccat caacgagaac caccgcaagt 6481 acgtggagaa ccacaagaag gagttcgagg agctgttcta ctacatcctg gagttcaacg 6541 agaactacgt gggcgccaag aagaacggca agctgctgaa ctcggccttc cagtcgtggc 6601 agaaccactc gatcgacgag ctgtgctcgt cgttcatcgg cccgaccggc tcggagcgca 6661 agggcctgtt cgagctgacc tcgcgcggct cggccgccga cttcgagttc ctgggcgtga 6721 agatcccgcg ctaccgcgac tacaccccgt cgtcgctgct gaaggacgcc accctgatcc 6781 accagtcggt gaccggcctg tacgagaccc gcatcgacct ggccaagctg ggcgagggct 6841 aaatactctg gccatttttc tttaccgcct ttgagtgagc tgataccgct cgccgcagcc 6901 gaacgaccga gcgcaacgcg tgcggccgct gattagctaa gcagaaggcc atcctgacgg 6961 atgccttttt cgcttgctag ctgcagcccg ggggatccac tagttctaga gcggccattc 7021 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 7081 ccttgcagca catccccctt tcgccagcgg tatcagctca ctcaaaggcg gtaatacggt 7141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 7861 agtggaacga aaactcacgt taagggattt tggtcatacg aggccctttc gtcttcaaga 7921 attaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 7981 cttaatcgcc ttgcagcaca tccccctttc gccagcagat ccgtccctat cgtttccacg 8041 atcagcgatc ggctcgttgc cctgcgccgc tccaaagccc gcgacgcagc gccggcaggc 8101 agagcaagta gagggcagcg cctgcaatcc atgcccaccc gttccacgtt gttatagaag 8161 ccgcatagat cgccgtgaag aggaggggtc cgacgatcga ggtcaggctg gtgagcgccg 8221 ccagtgagcc ttgcagctgc ccctgacgtt cctcatccac ctgcctggac aacattgctt 8281 gcagcgccgg cattccgatg ccacccgaag caagcaggac catgatcggg aacgccatcc 8341 atccccgtgt cgcgaaggca agcaggatgt agcctgtgcc gtcggcaatc attccgagca 8401 tgagtgcccg cctttcgccg agccgggcgg ctacagggcc ggtgatcatt gcctgggcga 8461 gtgaatgcag aatgccaaat gcggcaagcg aaatgccgat cgtggtcgcg tcccagtgaa 8521 agcgatcctc gccgaaaatg acccaaagcg cggccggcac ctgtccgaca agttgcatga 8581 tgaagaagac cgccatcagg gcggcgacga cggtcatgcc ccgggcccac cggaacgaag 8641 cgagcgggtt gagagcctcc cggcgtaacg gccggcgttc gcctttgtgc gactccggca 8701 aaaggaaaca gcccgtcagg aaattgaggc cgttcaaggc tgccgcggcg aagaacggag 8761 cgtgggggga gaaaccgccc atcagcccac cgagcacagg tcccgcgacc atcccgaacc 8821 cgaaacaggc gctcatgaag ccgaagtgcc gcgcgcgctc atcgccatca gtgatatcgg 8881 caatataagc gccggctacc gccccagtcg ccccggtgat gccggccacg atccgcccga 8941 tatagagaac ccaaaggaaa ggcgctgtcg ccatgatggc gtagtcgaca gtggcgccgg 9001 ccagcgagac gagcaagatt ggccgccgcc cgaaacgatc tgacagcgcg cccagcacag 9061 gtgcgcaggc aaattgcacc aacgcataca gcgccagcag aatgccatag tgggcggtga 9121 cgtcgttcga gtgaaccaga tcgcgcagga ggcccggcag caccggcata atcaggccga 9181 tgccgacagc gtcgagcgcg acagtgctca gaattacgat caggggtatg ttgggtttca 9241 cgtctggcct ccggaccagc ctccgctggt ccgattgaac gcgcggattc tttatcactg 9301 ataagttggt ggacatatta tgtttatcag tgataaagtg tcaagcatga caaagttgca 9361 gccgaataca gtgatccgtg ccgccctgga cctgttgaac gaggtcggcg tagacggtct 9421 gacgacacgc aaactggcgg aacggttggg ggttcagcag ccggcgcttt actggcactt 9481 caggaacaag cgggcgctgc tcgacgcact ggccgaagcc atgctggcgg agaatcatac 9541 gcattcggtg ccgagagccg acgacgactg gcgctcattt ctgatcggga atgcccgcag 9601 cttcaggcag gcgctgctcg cctaccgcga tggcgcgcgc atccatgccg gcacgcgacc 9661 gggcgcaccg cagatggaaa cggccgacgc gcagcttcgc ttcctctgcg aggcgggttt 9721 ttcggccggg gacgccgtca atgcgctgat gacaatcagc tacttcactg ttggggccgt 9781 gcttgaggag caggccggcg acagcgatgc cggcgagcgc ggcggcaccg ttgaacaggc 9841 tccgctctcg ccgctgttgc gggccgcgat agacgccttc gacgaagccg gtccggacgc 9901 agcgttcgag cagggactcg cggtgattgt cgatggattg gcgaaaagga ggctcgttgt 9961 caggaacgtt gaaggaccga gaaagggtga cgattgatca ggaccgctgc cggagcgcaa 10021 cccactcact acagcagagc catgtagaca acatcccctc cccctttcca ccgcgtcaga 10081 cgcccgtagc agcccgctac gggctttttc atgccctgcc ctagcgtcca agcctcacgg 10141 ccgcgctcgg cctctctggc ggccttctgg cgctcctgct gcggcgtccg ctcgtatgat 10201 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc 10261 cccccccccc ccaccgcggt ggcggccgcc ccggccagcc tcgcagagca ggattcccgt 10321 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc 10381 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa 10441 ccctttggca aaatcctgta tat //