LOCUS       sequence-269501-        3581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 8 to 42)
  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 3581)
  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: pCAT219
FEATURES             Location/Qualifiers
     source          1..3581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..42
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     CDS             73..750
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      783..854
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      870..897
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(971..1786)
                     /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(971..1786)
                     /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     1694..1713
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1778..1797
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2033..2804
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2805..3467
                     /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 tctctatttg acggctagct cagtcctagg tacagtgcta gcgtacttta actttaagaa
       61 ggagatatac atatggcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt
      121 cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt
      181 ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc
      241 gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg
      301 gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt
      361 atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt
      421 gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg
      481 cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct
      541 ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa
      601 gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac
      661 atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt
      721 gctgaaggtc gtcactccac cggtgcttaa ggatccaaac tcgagtaagg atctccaggc
      781 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      841 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
      901 aaagcggcgc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
      961 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1021 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1081 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1141 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1201 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
     1261 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1321 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1381 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1441 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1501 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1561 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1621 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1681 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1741 ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     1801 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     1861 tgtaacatca gagattttga gacacaacgt ggctttccag gtggccggcc cttcgcaaag
     1921 tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc ttcgccctgc
     1981 gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc gagcggccac
     2041 cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga tggacaggct
     2101 gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc ttcgaccaca
     2161 tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt ttaattttct
     2221 ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg acaccaagtg
     2281 gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac gcgcctggaa
     2341 cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc ctgccccccg
     2401 agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg gcaaggccga
     2461 aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag ggggagccgc
     2521 gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa gaactagata
     2581 tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt acgcaacagc
     2641 tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct gtaattgact
     2701 gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag ctgattgcgc
     2761 atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc acgcagtcca
     2821 gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg gaacgcaaag
     2881 cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg ctgctgcatc
     2941 acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag acactttcca
     3001 agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg gtggccgagc
     3061 gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac gtggtcaatg
     3121 accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc agtgccgccg
     3181 tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac ctgcgccgca
     3241 tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag gagccgccca
     3301 gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa acggaggaat
     3361 gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt ctggacgatg
     3421 gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac ttgggttgcg
     3481 cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc cgccctatac
     3541 cttgtctgga acttaattaa gccgcttcta gaacgtctca a
//
