LOCUS       sequence-266830-        9731 bp    DNA     circular SYN 15-DEC-2025
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    .
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. 2020 Jan 14;11(1):e02415-19. doi: 10.1128/mBio.02415-19.
  PUBMED    31937638
REFERENCE   2  (bases 1 to 9731)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1128/mBio"; 
            journalName: "mBio"; date: "2020-01-14- 14"; volume: "11"; issue: 
            "1"; pages: "e02415-19"
COMMENT     Sequence Label: pVCERC-5 Pvan-dcas9 (Sth3)
FEATURES             Location/Qualifiers
     source          1..9731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
//
