LOCUS       sequence-269499-        4153 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 4153)
  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 4153)
  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: pCAT207
FEATURES             Location/Qualifiers
     source          1..4153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             95..772
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      805..876
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      892..919
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     1065..1083
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1110..1115,1116..1132,1133..1138)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1186..2376
                     /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(1538..1557)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      2619..3390
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3391..4053
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 ataaaaaccg ttattgacac aggtggaaat ttagaataac ttgttagtaa acctaatgga
       61 tcgaccttag atcttttaag aaggagatat acatatggcg agtagcgaag acgttatcaa
      121 agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg agttcgaaat
      181 cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag accgctaaac tgaaagttac
      241 caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc agtacggctc
      301 caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt ccttcccgga
      361 aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta ccgttaccca
      421 ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt
      481 cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg
      541 tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga
      601 cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac cggttcagct
      661 gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg aagactacac
      721 catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt aaggatccaa
      781 actcgagtaa ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      841 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
      901 tcgggtgggc ctttctgcga agcggcgcgc cgttgggaag ccctgcaaag taaactggat
      961 ggctttcttg ccgccaagga tctgatggcg caggggatca agatctccga aaagtgccac
     1021 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1081 ggccctttcg tcttcaagaa ttctcatgtt tgacagctta tcatcgataa gctttaatgc
     1141 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat
     1201 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg
     1261 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat
     1321 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca
     1381 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc
     1441 gactacgcga tcatggcgac cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg
     1501 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     1561 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     1621 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     1681 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     1741 aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     1801 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     1861 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     1921 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     1981 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     2041 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     2101 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     2161 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc
     2221 ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     2281 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg
     2341 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta
     2401 acggattcac cactccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2461 ttttctggat tcatcgactg tggcaggtgg ccggcccttc gcaaagtcgt gaccgcctac
     2521 ggcggctgcg gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg tcgctgcgct
     2581 cccttgccag cccgtggata tgtggacgat ggccgcgagc ggccaccggc tggctcgctt
     2641 cgctcggccc gtggacaacc ctgctggaca agctgatgga caggctgcgc ctgcccacga
     2701 gcttgaccac agggattgcc caccggctac ccagccttcg accacatacc caccggctcc
     2761 aactgcgcgg cctgcggcct tgccccatca atttttttaa ttttctctgg ggaaaagcct
     2821 ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag gcgggtcaag
     2881 gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac ccaagcctat
     2941 gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc
     3001 gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg
     3061 atcaacaagc cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg
     3121 ggaaccccgc aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg
     3181 aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc
     3241 cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca
     3301 acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc
     3361 gtgcggcacc cctaccgcat ggagataagc atggccacgc agtccagaga aatcggcatt
     3421 caagccaaga acaagcccgg tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg
     3481 gccgggctta ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct cgtggcgcag
     3541 atgggccacc agaacgccgt ggtggtcagc cagaagacac tttccaagct catcggacgt
     3601 tctttgcgga cggtccaata cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc
     3661 gtgaagctca acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg
     3721 ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac
     3781 gacgaccagg acgaatcgct gttggggcat ggcgacctgc gccgcatccc gaccctgtat
     3841 ccgggcgagc agcaactacc gaccggcccc ggcgaggagc cgcccagcca gcccggcatt
     3901 ccgggcatgg aaccagacct gccagccttg accgaaacgg aggaatggga acggcgcggg
     3961 cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg acgatggcga gccgttggag
     4021 ccgccgacac gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag
     4081 tgcgctgttc cagactatcg gctgtagccg cctcgccgcc ctataccttg tctggaactt
     4141 aattaaagcg gat
//
