LOCUS       sequence-420113-        4580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Burkholderia multivorans LRD-modifying cytochrome P450
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4580)
  AUTHORS   Lin GM, Voigt CA
  TITLE     Design of a redox-proficient Escherichia coli for screening 
            terpenoids and modifying cytochrome P450s
  JOURNAL   Nature Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5
REFERENCE   2  (bases 1 to 4580)
  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: "Nature 
            Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5"
COMMENT     Sequence Label: pCYP05
FEATURES             Location/Qualifiers
     source          1..4580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(296..841)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(376..395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(930..1745)
                     /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(930..1745)
                     /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     1653..1672
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1737..1756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1922..1949
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    1980..2007
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     RBS             2099..2107
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(4492..4511)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 actgactcgc tgcacgaggc agacctcagc gctagcggag tgtatactgg cttactatgt
       61 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaacg
      121 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
      181 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
      241 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
      301 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
      361 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
      421 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
      481 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
      541 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      601 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
      661 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
      721 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
      781 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
      841 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacct tagaaaaact catcgagcat caaatgaaac
      961 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1021 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1081 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1141 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1201 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1261 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1321 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1381 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1441 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1501 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1561 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1621 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1681 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     1741 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1801 atatttttat cttgtgcaat gtacatcaga gattttgaga cacaaccaat tattgaaggc
     1861 ctccctaacg gggggccttt ttttgtttct ggtctcccaa gcgcttaacg atcgttggct
     1921 gaacaaacag acaatctggt ctgtttgtat tatggaaaat ttttctgtat aatagattca
     1981 acaaacagac aatctggtct gtttgtatta tcagcggtca acgcatgtgc tttgcgttct
     2041 gatgagacag tgatgtcgaa accgcctcta caaataattt tgtttagagc cagcactata
     2101 aggaggttat tgatgctgct gaacccgtta agtcgtcgtg tgcgcatgca gcaaggcatg
     2161 cccgtgttac cgggcgcgtt cccggtcgta ggccatctgc ccgctgttgc gcgcgcactg
     2221 cctgatctgc tggcatacgg cgaacgtaga tggggcaccc atttctggtt ggacatgggc
     2281 gcgatgggtc gtatgctggt gtgcacccgc cccgaggcct ttgaactgtt caacaacaaa
     2341 gacgtgctcg ccgatttact gggagatgtc gctccggcca tgcgtgacac gatggccacg
     2401 caagatggtg aactgcatcg tcgtacccgc gcttgggcga acccatcttt tcaaccacag
     2461 ggtcttagtg ccgccaaggt tggcgaactg ttcgcggata taatgatggc gtggattcgt
     2521 gcttggccag cgcgcggtac aattaacttg gtggacgaaa cgagcgcttt gacgctgaca
     2581 atgatcttcc gtatgttagg gatcgcagag gatgacattg caacgtggga aacccattat
     2641 cgccattatg cgctggtgag cctgcttccc gtggacctgc cgggtacgcc tttacgccgc
     2701 agccgtcgcg cacaagtctg gattgatgcg cgtctgtcag cattggtaaa cgaattgggt
     2761 cgtagcccag gcgactcggg cctcttagcg accctgatcc gttcacccgc ggccccggat
     2821 gcgtcgtcac cagaatcaaa tcttttaggt aacctgcgtg cactcattca cgccgcgcac
     2881 aaaaccaccg cggtaacgat ggctcacttg gcgatccgcc tcgcagagca gccagaccac
     2941 tgggatgcat tagtcgcaga agctcgggct gctggaacgg tgccgcgcaa cccctcggaa
     3001 ctggcacgtt tcccgttcgc tgaggccctc tttctggaaa cgctgcgctc tgagccggca
     3061 tttcccttag tgtttcgtcg cacccgcgaa cgaattgagc tgggcggtcg cgagctggct
     3121 gcaggtacac ggctgattat ccctctgatc ttgttatcgc gtgcgccaag cctgtacccg
     3181 cagccagata ctttcttacc gcagcgctgg ttagagcacc gaggcgccca taaaccaatt
     3241 cagactttac agttcggaag cggtccacat cgttgtctgg gttttcatac tgcctgcatg
     3301 caactggtgc agtttgcagt taccttggcc ctggtgctgg atgaacggaa aatgcgcccg
     3361 caacttgtgc cgcagcggac ttgtgggcgc cgtcgttttc cgattaacta tccaccacgt
     3421 gacacccagg tggcctttag aagtgtttct tcgaatcacg acaaccgtag tgagattccg
     3481 agctaataag gatgagctct acaaataagc agaggtggtt gtgttgcgaa aaaaaaaaaa
     3541 aaacacccta acgggtgttt tttttttttt ggtgtcccca cgtgtggcgc tggagaccgt
     3601 ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     3661 ggaagagagt caattcaggg tggtgaatat gagcccgaaa cgtcgtaccc aggcagaacg
     3721 tgcaatggaa acccagggta aactgattgc agcagcactg ggtgttctgc gtgaaaaagg
     3781 ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc ggtgttagcc gtggtgcaca
     3841 gagccatcat tttccgacca aactggaact gctgctggca acctttgaat ggctgtatga
     3901 gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg aaaccggaag atgatgttat
     3961 tcagcagatg ctggatgatg cagcagattt ttttctggat gatgatttta gcatcggcct
     4021 ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt gaaggtattc tgcgtaccgt
     4081 tgaacgtaat cgttttgttg ttgaagatat gtggctgggt gtgctggtga gccgtggtct
     4141 gagccgtgat gatgccgaag atattctgtg gctgattttt aacagcgttc gtggtctgac
     4201 agttcgtagc ctgtggcaga aagataaaga acgttttgaa cgtgtgcgta atagcaccct
     4261 ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga taaggatcct aattggtaac
     4321 gaatcagaca attgacggct cgagggagta gcatagggtt tgcagaatcc ctgcttcgtc
     4381 catttgacag gcacattatg catcgatgat aagctgtcaa acatgagcag atcctctacg
     4441 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     4501 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc aaatatttta
     4561 tctgaggtgc ttcctcgctc
//
