LOCUS       sequence-266835-       10463 bp    DNA     circular SYN 15-DEC-2025
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    .
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. 2020 Jan 14;11(1):e02415-19. doi: 10.1128/mBio.02415-19.
  PUBMED    31937638
REFERENCE   2  (bases 1 to 10463)
  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: pXGFPC-5 Pxyl-dcas9 (Sth3)
FEATURES             Location/Qualifiers
     source          1..10463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
//
