LOCUS       sequence-269067-        3556 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 3556)
  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 3556)
  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: pCAT202
FEATURES             Location/Qualifiers
     source          1..3556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             101..775
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDTIVEQYERAEGRHST
                     GA"
     terminator      808..879
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      895..922
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(992..1807)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(992..1807)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1715..1734
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1799..1818
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2054..2825
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2826..3488
                     /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 tataccttgt ctggaactta attaaagcgg atataaaaac cgttattgac acaggtggaa
       61 atttagaata acttgttagt aaacctaatg gatcgacctt atggcgagta gcgaagacgt
      121 tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt
      181 cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa
      241 agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc ctgtccccgc agttccagta
      301 cggttccaaa gcttacgtta aacacccggc tgacatcccg gactacctga aactgtcctt
      361 cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt
      421 tacccaggac tcctccctgc aagacggtga gttcatctac aaagttaaac tgcgtggtac
      481 caacttcccg tccgacggtc cggttatgca gaaaaaaacc atgggttggg aagcttccac
      541 cgaacgtatg tacccggaag acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact
      601 gaaagacggt ggtcactacg acgctgaagt taaaaccacc tacatggcta aaaaaccggt
      661 tcagctgccg ggtgcttaca aaaccgacat caaactggac atcacctccc acaacgaaga
      721 caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaaggatc
      781 caaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      841 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
      901 ccttcgggtg ggcctttctg cgaagcggcg cgccgtcccg tcaagtcagc gtaatgctct
      961 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     1021 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1081 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1141 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1201 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     1261 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1321 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     1381 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1441 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     1501 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1561 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1621 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1681 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1741 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
     1801 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     1861 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttcca
     1921 ggtggccggc ccttcgcaaa gtcgtgaccg cctacggcgg ctgcggcgcc ctacgggctt
     1981 gctctccggg cttcgccctg cgcggtcgct gcgctccctt gccagcccgt ggatatgtgg
     2041 acgatggccg cgagcggcca ccggctggct cgcttcgctc ggcccgtgga caaccctgct
     2101 ggacaagctg atggacaggc tgcgcctgcc cacgagcttg accacaggga ttgcccaccg
     2161 gctacccagc cttcgaccac atacccaccg gctccaactg cgcggcctgc ggccttgccc
     2221 catcaatttt tttaattttc tctggggaaa agcctccggc ctgcggcctg cgcgcttcgc
     2281 ttgccggttg gacaccaagt ggaaggcggg tcaaggctcg cgcagcgacc gcgcagcggc
     2341 ttggccttga cgcgcctgga acgacccaag cctatgcgag tgggggcagt cgaagggcga
     2401 agcccgcccg cctgcccccc gagcctcacg gcggcgagtg cgggggttcc aagggggcag
     2461 cgccaccttg ggcaaggccg aaggccgcgc agtcgatcaa caagccccgg aggggccact
     2521 ttttgccgga gggggagccg cgccgaaggc gtgggggaac cccgcagggg tgcccttctt
     2581 tgggcaccaa agaactagat atagggcgaa atgcgaaaga cttaaaaatc aacaacttaa
     2641 aaaagggggg tacgcaacag ctcattgcgg caccccccgc aatagctcat tgcgtaggtt
     2701 aaagaaaatc tgtaattgac tgccactttt acgcaacgca taattgttgt cgcgctgccg
     2761 aaaagttgca gctgattgcg catggtgccg caaccgtgcg gcacccctac cgcatggaga
     2821 taagcatggc cacgcagtcc agagaaatcg gcattcaagc caagaacaag cccggtcact
     2881 gggtgcaaac ggaacgcaaa gcgcatgagg cgtgggccgg gcttattgcg aggaaaccca
     2941 cggcggcaat gctgctgcat cacctcgtgg cgcagatggg ccaccagaac gccgtggtgg
     3001 tcagccagaa gacactttcc aagctcatcg gacgttcttt gcggacggtc caatacgcag
     3061 tcaaggactt ggtggccgag cgctggatct ccgtcgtgaa gctcaacggc cccggcaccg
     3121 tgtcggccta cgtggtcaat gaccgcgtgg cgtggggcca gccccgcgac cagttgcgcc
     3181 tgtcggtgtt cagtgccgcc gtggtggttg atcacgacga ccaggacgaa tcgctgttgg
     3241 ggcatggcga cctgcgccgc atcccgaccc tgtatccggg cgagcagcaa ctaccgaccg
     3301 gccccggcga ggagccgccc agccagcccg gcattccggg catggaacca gacctgccag
     3361 ccttgaccga aacggaggaa tgggaacggc gcgggcagca gcgcctgccg atgcccgatg
     3421 agccgtgttt tctggacgat ggcgagccgt tggagccgcc gacacgggtc acgctgccgc
     3481 gccggtagca cttgggttgc gcagcaaccc gtaagtgcgc tgttccagac tatcggctgt
     3541 agccgcctcg ccgccc
//
