LOCUS       sequence-269075-        3551 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 3551)
  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 3551)
  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: pCAT217
FEATURES             Location/Qualifiers
     source          1..3551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(44..49,50..65,66..72)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             111..788
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      821..892
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      908..935
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1005..1820)
                     /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(1005..1820)
                     /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     1728..1747
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1812..1831
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2067..2838
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2839..3501
                     /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 gtagccgcct cgccgcccta taccttgtct ggaacttaat taattgacaa ttaatcatcg
       61 gctcgtataa tgtgtggaat tgtgagatct tttaagaagg agatatacat atggcgagta
      121 gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt tccgttaacg
      181 gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt acccagaccg
      241 ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc ctgtccccgc
      301 agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg gactacctga
      361 aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa gacggtggtg
      421 ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac aaagttaaac
      481 tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc atgggttggg
      541 aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa atcaaaatgc
      601 gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc tacatggcta
      661 aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac atcacctccc
      721 acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt cactccaccg
      781 gtgcttaagg atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca
      841 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
      901 tcacactggc tcaccttcgg gtgggccttt ctgcgaagcg gcgcgccgtc ccgtcaagtc
      961 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     1021 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1081 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1141 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1201 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1261 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1321 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1381 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1441 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1501 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1561 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1621 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1681 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1741 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt
     1801 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     1861 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     1921 acgtggcttt ccaggtggcc ggcccttcgc aaagtcgtga ccgcctacgg cggctgcggc
     1981 gccctacggg cttgctctcc gggcttcgcc ctgcgcggtc gctgcgctcc cttgccagcc
     2041 cgtggatatg tggacgatgg ccgcgagcgg ccaccggctg gctcgcttcg ctcggcccgt
     2101 ggacaaccct gctggacaag ctgatggaca ggctgcgcct gcccacgagc ttgaccacag
     2161 ggattgccca ccggctaccc agccttcgac cacataccca ccggctccaa ctgcgcggcc
     2221 tgcggccttg ccccatcaat ttttttaatt ttctctgggg aaaagcctcc ggcctgcggc
     2281 ctgcgcgctt cgcttgccgg ttggacacca agtggaaggc gggtcaaggc tcgcgcagcg
     2341 accgcgcagc ggcttggcct tgacgcgcct ggaacgaccc aagcctatgc gagtgggggc
     2401 agtcgaaggg cgaagcccgc ccgcctgccc cccgagcctc acggcggcga gtgcgggggt
     2461 tccaaggggg cagcgccacc ttgggcaagg ccgaaggccg cgcagtcgat caacaagccc
     2521 cggaggggcc actttttgcc ggagggggag ccgcgccgaa ggcgtggggg aaccccgcag
     2581 gggtgccctt ctttgggcac caaagaacta gatatagggc gaaatgcgaa agacttaaaa
     2641 atcaacaact taaaaaaggg gggtacgcaa cagctcattg cggcaccccc cgcaatagct
     2701 cattgcgtag gttaaagaaa atctgtaatt gactgccact tttacgcaac gcataattgt
     2761 tgtcgcgctg ccgaaaagtt gcagctgatt gcgcatggtg ccgcaaccgt gcggcacccc
     2821 taccgcatgg agataagcat ggccacgcag tccagagaaa tcggcattca agccaagaac
     2881 aagcccggtc actgggtgca aacggaacgc aaagcgcatg aggcgtgggc cgggcttatt
     2941 gcgaggaaac ccacggcggc aatgctgctg catcacctcg tggcgcagat gggccaccag
     3001 aacgccgtgg tggtcagcca gaagacactt tccaagctca tcggacgttc tttgcggacg
     3061 gtccaatacg cagtcaagga cttggtggcc gagcgctgga tctccgtcgt gaagctcaac
     3121 ggccccggca ccgtgtcggc ctacgtggtc aatgaccgcg tggcgtgggg ccagccccgc
     3181 gaccagttgc gcctgtcggt gttcagtgcc gccgtggtgg ttgatcacga cgaccaggac
     3241 gaatcgctgt tggggcatgg cgacctgcgc cgcatcccga ccctgtatcc gggcgagcag
     3301 caactaccga ccggccccgg cgaggagccg cccagccagc ccggcattcc gggcatggaa
     3361 ccagacctgc cagccttgac cgaaacggag gaatgggaac ggcgcgggca gcagcgcctg
     3421 ccgatgcccg atgagccgtg ttttctggac gatggcgagc cgttggagcc gccgacacgg
     3481 gtcacgctgc cgcgccggta gcacttgggt tgcgcagcaa cccgtaagtg cgctgttcca
     3541 gactatcggc t
//
