LOCUS       sequence-273743-        3165 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 3165)
  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 3165)
  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: pCAT204
FEATURES             Location/Qualifiers
     source          1..3165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        613..715
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             716..1375
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(782..801)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        join(1540..1545,1546..1563,1564..1570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1578..1594
                     /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)."
     primer_bind     1583..1605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1602..1618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1602..1618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1614..1979
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     primer_bind     1637..1657
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1639..1657
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1670..1777
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1686..1702
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1687..1703
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1737..1753)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1737..1753)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1785..1804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1786..1804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1814..1831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1814..1830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1823..1845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2153..2815)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      join(2816..3165,1..422)
                     /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 ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga ggctcggggg
       61 gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg gtcgttccag
      121 gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg
      181 gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa
      241 ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt
      301 cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg
      361 tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc
      421 gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg
      481 cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc
      541 gaagggccgg ccacctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
      601 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
      661 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
      721 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
      781 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
      841 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
      901 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
      961 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     1021 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1081 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1141 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     1201 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     1261 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     1321 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     1381 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa
     1441 gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt
     1501 cgttaaggcg cgccgcttca ctcattaggc accccaggct ttacacttta tgcttccggc
     1561 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1621 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg gtaccgggcc
     1681 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
     1741 cactagttct agagcggccg ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt
     1801 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1861 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1921 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg aaattgtaag
     1981 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2041 ataggccgac tgcgatgagt taattaagtt ccagacaagg tatagggcgg cgaggcggct
     2101 acagccgata gtctggaaca gcgcacttac gggttgctgc gcaacccaag tgctaccggc
     2161 gcggcagcgt gacccgtgtc ggcggctcca acggctcgcc atcgtccaga aaacacggct
     2221 catcgggcat cggcaggcgc tgctgcccgc gccgttccca ttcctccgtt tcggtcaagg
     2281 ctggcaggtc tggttccatg cccggaatgc cgggctggct gggcggctcc tcgccggggc
     2341 cggtcggtag ttgctgctcg cccggataca gggtcgggat gcggcgcagg tcgccatgcc
     2401 ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac ggcggcactg aacaccgaca
     2461 ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc attgaccacg taggccgaca
     2521 cggtgccggg gccgttgagc ttcacgacgg agatccagcg ctcggccacc aagtccttga
     2581 ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt ggaaagtgtc ttctggctga
     2641 ccaccacggc gttctggtgg cccatctgcg ccacgaggtg atgcagcagc attgccgccg
     2701 tgggtttcct cgcaataagc ccggcccacg cctcatgcgc tttgcgttcc gtttgcaccc
     2761 agtgaccggg cttgttcttg gcttgaatgc cgatttctct ggactgcgtg gccatgctta
     2821 tctccatgcg gtaggggtgc cgcacggttg cggcaccatg cgcaatcagc tgcaactttt
     2881 cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca gtcaattaca gattttcttt
     2941 aacctacgca atgagctatt gcggggggtg ccgcaatgag ctgttgcgta cccccctttt
     3001 ttaagttgtt gatttttaag tctttcgcat ttcgccctat atctagttct ttggtgccca
     3061 aagaagggca cccctgcggg gttcccccac gccttcggcg cggctccccc tccggcaaaa
     3121 agtggcccct ccggggcttg ttgatcgact gcgcggcctt cggcc
//
