LOCUS       sequence-259187-        3557 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 3557)
  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 3557)
  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..3557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             88..765
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      798..869
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      885..912
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(982..1797)
                     /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(982..1797)
                     /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     1705..1724
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1789..1808
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2044..2813
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2814..3476
                     /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 gaacttaatt aaagcggata taaaaaccgt tattgacaca ggtggaaatt tagaataact
       61 tgttagtaaa cctaatggat cgaccttatg gcgagtagcg aagacgttat caaagagttc
      121 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
      181 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
      241 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
      301 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
      361 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
      421 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
      481 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
      541 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
      601 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
      661 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
      721 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaaggatc caaactcgag
      781 taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      841 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
      901 ggcctttctg cgaagcggcg cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta
      961 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1021 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1081 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1141 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1201 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
     1261 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1321 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     1381 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1441 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     1501 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1561 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1621 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     1681 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1741 catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat ggctcataac
     1801 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     1861 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttcca ggtggccggc
     1921 ccttcgcaaa gtcgtgaccg cctacggcgg ctgcggcgcc ctacgggctt gctctccggg
     1981 cttcgccctg cgcggtcgct gcgctccctt gccagcccgt ggatatgtgg acgatggccg
     2041 cgagcggcca ccggctggct cgcttcgctc ggcccgtgga caaccctgct ggacaagctg
     2101 atggacaggc tgcgcctgcc cacgagcttg accacaggga ttgcccaccg gctacccagc
     2161 cttcgaccac atacccaccg gctccaactg cgcggcctgc ggccttgccc catcaatttt
     2221 tttaattttc tctggggaaa agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg
     2281 gacaccaagt ggaaggcggg tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga
     2341 cgcgcctgga acgacccaag cctatgcgag tgggggcagt cgaaggcgaa gcccgcccgc
     2401 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     2461 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     2521 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     2581 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     2641 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     2701 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     2761 ctgattgcgc atggtgccgc aaccgtgcgg caccctaccg catggagata agcatggcca
     2821 cgcagtccag agaaatcggc attcaagcca agaacaagcc cggtcactgg gtgcaaacgg
     2881 aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag gaaacccacg gcggcaatgc
     2941 tgctgcatca cctcgtggcg cagatgggcc accagaacgc cgtggtggtc agccagaaga
     3001 cactttccaa gctcatcgga cgttctttgc ggacggtcca atacgcagtc aaggacttgg
     3061 tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg
     3121 tggtcaatga ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg tcggtgttca
     3181 gtgccgccgt ggtggttgat cacgacgacc aggacgaatc gctgttgggg catggcgacc
     3241 tgcgccgcat cccgaccctg tatccgggcg agcagcaact accgaccggc cccggcgagg
     3301 agccgcccag ccagcccggc attccgggca tggaaccaga cctgccagcc ttgaccgaaa
     3361 cggaggaatg ggaacggcgc gggcagcagc gcctgccgat gcccgatgag ccgtgttttc
     3421 tggacgatgg cgagccgttg gagccgccga cacgggtcac gctgccgcgc cggtagcact
     3481 tgggttgcgc agcaacccgt aagtgcgctg ttccagacta tcggctgtag ccgcctcgcc
     3541 gccctatacc ttgtctg
//
