LOCUS       sequence-259195-        3579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 41 to 75)
  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 3579)
  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..3579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..75
                     /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             106..783
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      816..887
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      903..930
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1004..1819)
                     /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(1004..1819)
                     /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     1727..1746
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1811..1830
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2066..2835
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2836..3498
                     /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 aagccgcttc tagaacgtct caatctctat ttgacggcta gctcagtcct
       61 aggtacagtg ctagcgtact ttaactttaa gaaggagata tacatatggc gagtagcgaa
      121 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
      181 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
      241 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
      301 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
      361 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
      421 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
      481 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
      541 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
      601 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
      661 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
      721 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
      781 taaggatcca aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
      841 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
      901 ctggctcacc ttcgggtggg cctttctgcg tttaaagcgg cgcgccgtcc cgtcaagtca
      961 gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     1021 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1081 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1141 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1201 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1261 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1321 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1381 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     1441 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     1501 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     1561 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     1621 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     1681 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     1741 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     1801 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     1861 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     1921 cgtggctttc caggtggccg gcccttcgca aagtcgtgac cgcctacggc ggctgcggcg
     1981 ccctacgggc ttgctctccg ggcttcgccc tgcgcggtcg ctgcgctccc ttgccagccc
     2041 gtggatatgt ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg
     2101 gacaaccctg ctggacaagc tgatggacag gctgcgcctg cccacgagct tgaccacagg
     2161 gattgcccac cggctaccca gccttcgacc acatacccac cggctccaac tgcgcggcct
     2221 gcggccttgc cccatcaatt tttttaattt tctctgggga aaagcctccg gcctgcggcc
     2281 tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga
     2341 ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca agcctatgcg agtgggggca
     2401 gtcgaaggcg aagcccgccc gcctgccccc cgagcctcac ggcggcgagt gcgggggttc
     2461 caagggggca gcgccacctt gggcaaggcc gaaggccgcg cagtcgatca acaagccccg
     2521 gaggggccac tttttgccgg agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg
     2581 gtgcccttct ttgggcacca aagaactaga tatagggcga aatgcgaaag acttaaaaat
     2641 caacaactta aaaaaggggg gtacgcaaca gctcattgcg gcaccccccg caatagctca
     2701 ttgcgtaggt taaagaaaat ctgtaattga ctgccacttt tacgcaacgc ataattgttg
     2761 tcgcgctgcc gaaaagttgc agctgattgc gcatggtgcc gcaaccgtgc ggcaccctac
     2821 cgcatggaga taagcatggc cacgcagtcc agagaaatcg gcattcaagc caagaacaag
     2881 cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg cgtgggccgg gcttattgcg
     2941 aggaaaccca cggcggcaat gctgctgcat cacctcgtgg cgcagatggg ccaccagaac
     3001 gccgtggtgg tcagccagaa gacactttcc aagctcatcg gacgttcttt gcggacggtc
     3061 caatacgcag tcaaggactt ggtggccgag cgctggatct ccgtcgtgaa gctcaacggc
     3121 cccggcaccg tgtcggccta cgtggtcaat gaccgcgtgg cgtggggcca gccccgcgac
     3181 cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg atcacgacga ccaggacgaa
     3241 tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc tgtatccggg cgagcagcaa
     3301 ctaccgaccg gccccggcga ggagccgccc agccagcccg gcattccggg catggaacca
     3361 gacctgccag ccttgaccga aacggaggaa tgggaacggc gcgggcagca gcgcctgccg
     3421 atgcccgatg agccgtgttt tctggacgat ggcgagccgt tggagccgcc gacacgggtc
     3481 acgctgccgc gccggtagca cttgggttgc gcagcaaccc gtaagtgcgc tgttccagac
     3541 tatcggctgt agccgcctcg ccgccctata ccttgtctg
//
