LOCUS       sequence-269497-        3550 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 3550)
  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 3550)
  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: pCAT205
FEATURES             Location/Qualifiers
     source          1..3550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             183..860
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      893..964
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      980..1007
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1151..1810)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1725..1744
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1811..1913)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2104..2875
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2876..3538
                     /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 gcagcaaccc gtaagtgcgc tgttccagac tatcggctgt agccgcctcg ccgccctata
       61 ccttgtctgg aacttaatta aagcggatat aaaaaccgtt attgacacag gtggaaattt
      121 agaataactt gttagtaaac ctaatggatc gaccttagat cttttaagaa ggagatatac
      181 atatggcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
      241 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
      301 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
      361 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
      421 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
      481 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
      541 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
      601 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
      661 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
      721 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
      781 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
      841 gtcactccac cggtgcttaa ggatccaaac tcgagtaagg atctccaggc atcaaataaa
      901 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      961 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgaag cggcgcgcct
     1021 taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca
     1081 tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac caataactgc
     1141 cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     1201 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     1261 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     1321 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     1381 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     1441 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     1501 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     1561 tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg
     1621 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     1681 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     1741 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     1801 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     1861 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
     1921 attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca ggtggccggc
     1981 ccttcgcaaa gtcgtgaccg cctacggcgg ctgcggcgcc ctacgggctt gctctccggg
     2041 cttcgccctg cgcggtcgct gcgctccctt gccagcccgt ggatatgtgg acgatggccg
     2101 cgagcggcca ccggctggct cgcttcgctc ggcccgtgga caaccctgct ggacaagctg
     2161 atggacaggc tgcgcctgcc cacgagcttg accacaggga ttgcccaccg gctacccagc
     2221 cttcgaccac atacccaccg gctccaactg cgcggcctgc ggccttgccc catcaatttt
     2281 tttaattttc tctggggaaa agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg
     2341 gacaccaagt ggaaggcggg tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga
     2401 cgcgcctgga acgacccaag cctatgcgag tgggggcagt cgaagggcga agcccgcccg
     2461 cctgcccccc gagcctcacg gcggcgagtg cgggggttcc aagggggcag cgccaccttg
     2521 ggcaaggccg aaggccgcgc agtcgatcaa caagccccgg aggggccact ttttgccgga
     2581 gggggagccg cgccgaaggc gtgggggaac cccgcagggg tgcccttctt tgggcaccaa
     2641 agaactagat atagggcgaa atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg
     2701 tacgcaacag ctcattgcgg caccccccgc aatagctcat tgcgtaggtt aaagaaaatc
     2761 tgtaattgac tgccactttt acgcaacgca taattgttgt cgcgctgccg aaaagttgca
     2821 gctgattgcg catggtgccg caaccgtgcg gcacccctac cgcatggaga taagcatggc
     2881 cacgcagtcc agagaaatcg gcattcaagc caagaacaag cccggtcact gggtgcaaac
     2941 ggaacgcaaa gcgcatgagg cgtgggccgg gcttattgcg aggaaaccca cggcggcaat
     3001 gctgctgcat cacctcgtgg cgcagatggg ccaccagaac gccgtggtgg tcagccagaa
     3061 gacactttcc aagctcatcg gacgttcttt gcggacggtc caatacgcag tcaaggactt
     3121 ggtggccgag cgctggatct ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta
     3181 cgtggtcaat gaccgcgtgg cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt
     3241 cagtgccgcc gtggtggttg atcacgacga ccaggacgaa tcgctgttgg ggcatggcga
     3301 cctgcgccgc atcccgaccc tgtatccggg cgagcagcaa ctaccgaccg gccccggcga
     3361 ggagccgccc agccagcccg gcattccggg catggaacca gacctgccag ccttgaccga
     3421 aacggaggaa tgggaacggc gcgggcagca gcgcctgccg atgcccgatg agccgtgttt
     3481 tctggacgat ggcgagccgt tggagccgcc gacacgggtc acgctgccgc gccggtagca
     3541 cttgggttgc
//
