LOCUS       sequence-270280-        3768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3768)
  AUTHORS   Azubuike CC, Gatehouse AMR, Howard TP
  TITLE     pCAT vectors overcome inefficient electroporation of Cupriavidus 
            necator H16.
  JOURNAL   N Biotechnol. 2021 Nov 25;65:20-30. doi: 10.1016/j.nbt.2021.07.003. 
            Epub 2021 Jul 29.
  PUBMED    34333160
REFERENCE   2  (bases 1 to 3768)
  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"; journalName: "N 
            Biotechnol."; date: "2021-11-25"; volume: "65:20-30. doi"; pages: " 
            10.1016/j.nbt.2021.07.003. Epub 2021 Jul 29"
COMMENT     Sequence Label: pCAT206
FEATURES             Location/Qualifiers
     source          1..3768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             651..1313
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(1487..1852)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     primer_bind     1621..1643
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1635..1652
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1636..1652
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1662..1681
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1662..1680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1689..1796
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1713..1729
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1713..1729
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1763..1779)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1764..1780)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1809..1829)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1809..1827)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1848..1864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1848..1864)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1861..1883)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1872..1888
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1896..1902,1903..1920,1921..1926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2093..2111
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2138..2143,2144..2160,2161..2166)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2214..3404
                     /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(2566..2585)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      join(3647..3768,1..650)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accacatacc caccggctcc aactgcgcgg cctgcggcct tgccccatca atttttttaa
       61 ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac
      121 caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc
      181 ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc
      241 cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa
      301 ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg ccggaggggg
      361 agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc accaaagaac
      421 tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc
      481 aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa
      541 ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga
      601 ttgcgcatgg tgccgcaacc gtgcggcacc cctaccgcat ggagataagc atggccacgc
      661 agtccagaga aatcggcatt caagccaaga acaagcccgg tcactgggtg caaacggaac
      721 gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg gcaatgctgc
      781 tgcatcacct cgtggcgcag atgggccacc agaacgccgt ggtggtcagc cagaagacac
      841 tttccaagct catcggacgt tctttgcgga cggtccaata cgcagtcaag gacttggtgg
      901 ccgagcgctg gatctccgtc gtgaagctca acggccccgg caccgtgtcg gcctacgtgg
      961 tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg
     1021 ccgccgtggt ggttgatcac gacgaccagg acgaatcgct gttggggcat ggcgacctgc
     1081 gccgcatccc gaccctgtat ccgggcgagc agcaactacc gaccggcccc ggcgaggagc
     1141 cgcccagcca gcccggcatt ccgggcatgg aaccagacct gccagccttg accgaaacgg
     1201 aggaatggga acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg
     1261 acgatggcga gccgttggag ccgccgacac gggtcacgct gccgcgccgg tagcacttgg
     1321 gttgcgcagc aacccgtaag tgcgctgttc cagactatcg gctgtagccg cctcgccgcc
     1381 ctataccttg tctggaactt aattaactca tcgcagtcgg cctattggtt aaaaaatgag
     1441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt
     1501 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     1561 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     1621 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     1681 gcgaattgga gctccaccgc ggtggcggcc gctctagaac tagtggatcc cccgggctgc
     1741 aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccagc
     1801 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     1861 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1921 tgtaaagcct ggggtgccta atgagtgaag cggcgcgccg ttgggaagcc ctgcaaagta
     1981 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctccgaaa
     2041 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     2101 tatcacgagg ccctttcgtc ttcaagaatt ctcatgtttg acagcttatc atcgataagc
     2161 tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat
     2221 ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta ggcataggct
     2281 tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac agcatcgcca
     2341 gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc gcacccgttc
     2401 tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg ctacttggag
     2461 ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggatcctc tacgccggac
     2521 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     2581 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     2641 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     2701 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     2761 agtcgcataa gggagagcgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2821 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2881 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     2941 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     3001 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     3061 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     3121 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     3181 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     3241 ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat gccgcctcgg
     3301 cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc
     3361 ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca
     3421 cctcgctaac ggattcacca ctccatggcg atgcctgctt gccgaatatc atggtggaaa
     3481 atggccgctt ttctggattc atcgactgtg gcaggtggcc ggcccttcgc aaagtcgtga
     3541 ccgcctacgg cggctgcggc gccctacggg cttgctctcc gggcttcgcc ctgcgcggtc
     3601 gctgcgctcc cttgccagcc cgtggatatg tggacgatgg ccgcgagcgg ccaccggctg
     3661 gctcgcttcg ctcggcccgt ggacaaccct gctggacaag ctgatggaca ggctgcgcct
     3721 gcccacgagc ttgaccacag ggattgccca ccggctaccc agccttcg
//
