LOCUS       sequence-484264-        3584 bp    DNA     circular SYN 21-AUG-2025
DEFINITION  Bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3584)
  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 3584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 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: pCAT203
FEATURES             Location/Qualifiers
     source          1..3584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             114..791
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      824..895
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      911..938
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1008..1823)
                     /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(1008..1823)
                     /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     1731..1750
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1815..1834
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2069..2840
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2841..3503
                     /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 cgaccttaga tcttttaaga aggagatata catatggcga
      121 gtagcgaaga cgttatcaaa gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta
      181 acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga
      241 ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc
      301 cgcagttcca gtacggttcc aaagcttacg ttaaacaccc ggctgacatc ccggactacc
      361 tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg
      421 gtgttgttac cgttacccag gactcctccc tgcaagacgg tgagttcatc tacaaagtta
      481 aactgcgtgg taccaacttc ccgtccgacg gtccggttat gcagaaaaaa accatgggtt
      541 gggaagcttc caccgaacgt atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa
      601 tgcgtctgaa actgaaagac ggtggtcact acgacgctga agttaaaacc acctacatgg
      661 ctaaaaaacc ggttcagctg ccgggtgctt acaaaaccga catcaaactg gacatcacct
      721 cccacaacga agactacacc atcgttgaac agtacgaacg tgctgaaggt cgtcactcca
      781 ccggtgctta aggatccaaa ctcgagtaag gatctccagg catcaaataa aacgaaaggc
      841 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
      901 gagtcacact ggctcacctt cgggtgggcc tttctgcgaa gcggcgcgcc gtcccgtcaa
      961 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     1021 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1081 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1141 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1201 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     1261 aatggcaaaa gyttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     1321 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     1381 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1441 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1501 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1561 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1621 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1681 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1741 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
     1801 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     1861 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     1921 acacgtggct ttccaggtgg ccggcccttc gcaaagtcgt gaccgcctac ggcggctgcg
     1981 gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg tcgctgcgct cccttgccag
     2041 cccgtggata tgtggacgat ggccgcgagc ggccaccggc tggctcgctt cgctcggccc
     2101 gtggacaacc ctgctggaca agctgatgga caggctgcgc ctgcccacga gcttgaccac
     2161 agggattgcc caccggctac ccagccttcg accacatacc caccggctcc aactgcgcgg
     2221 cctgcggcct tgccccatca atttttttaa ttttctctgg ggaaaagcct ccggcctgcg
     2281 gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag gcgggtcaag gctcgcgcag
     2341 cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg
     2401 gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg
     2461 gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc
     2521 cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc
     2581 aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg aaagacttaa
     2641 aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc cccgcaatag
     2701 ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca acgcataatt
     2761 gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc gtgcggcacc
     2821 cctaccgcat ggagataagc atggccacgc agtccagaga aatcggcatt caagccaaga
     2881 acaagcccgg tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg gccgggctta
     2941 ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct cgtggcgcag atgggccacc
     3001 agaacgccgt ggtggtcagc cagaagacac tttccaagct catcggacgt tctttgcgga
     3061 cggtccaata cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc gtgaagctca
     3121 acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc
     3181 gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg
     3241 acgaatcgct gttggggcat ggcgacctgc gccgcatccc gaccctgtat ccgggcgagc
     3301 agcaactacc gaccggcccc ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg
     3361 aaccagacct gccagccttg accgaaacgg aggaatggga acggcgcggg cagcagcgcc
     3421 tgccgatgcc cgatgagccg tgttttctgg acgatggcga gccgttggag ccgccgacac
     3481 gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc
     3541 cagactatcg gctgtagccg cctcgccgcc ctataccttg tctg
//
