LOCUS       sequence-424409-        4416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Bradyrhizobium sp. Rc3b LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4416)
  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 4416)
  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: pCYP12
FEATURES             Location/Qualifiers
     source          1..4416
                     /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)"
     primer_bind     complement(4328..4347)
                     /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 ggctgcaccg
      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 tgtttaactt aattttcccg
     2101 atccctaagg agtaatctaa tgagtgagca accagtcccg atgctgccga tgtggcgcgt
     2161 tgaccacatc aaaccatcgc cggaaatgct ggcgttatgt gccaacggcc cgattcaccg
     2221 tgtccgtttc ccctctggtc atgaaggctg gtgcgtgacc ggttatgacg aagctaaagc
     2281 cgcactgtca gatgccgcgt tccgtccggc tggcatgcct ccgaccgcgt ttacccccga
     2341 ttcagttatc ttgggcagcc ccgggtggct cgtttctcac gaaggtggtg aacatgttcg
     2401 tttacgtatg cttgtggcac cggcatttag ttcccgacgc atgaaactgt tggcgccaca
     2461 tgtggatgct atcgccgcgc aactgttcga aaatctggct gcacagcgtc aaccggctga
     2521 cttgcgtcgc catctgagct tcccgctgcc agccatggtc atcagcgccc tgatgggtct
     2581 gctgtatgag gatcacgcct tctttgcccg cctcagcgat caagtcatga cccatcagca
     2641 tgaatccgga ccacgctcgg cgagccgctt ggcttgggaa gaactgcgtg cgtatatccg
     2701 gggaaaaatg cgcgataagc gccaggaccc tggagacaac ttactgaccg acctgctcac
     2761 ggcggtggat cagggcaaag ccacggaaga agaagccata ggtttagccg ccggtatgct
     2821 ggtggccggt cacgaaagca cagttgcgca aattgagtat gggttactgg ccatgttctg
     2881 tcatcgtcag caacgcgagc gcctcgtagg cgacccgtcg ttattagata gcgccgtcga
     2941 agaaatcttg cgaatgtacc cgccgggagc agggtgggac ggcatcatgc gatatccgcg
     3001 aaccgacgtt accattgcag gcgtccatat tccagcagaa tccaaggttt tggttggtct
     3061 gccagctacg agcttcgacc ctcgccattt cgacgagcct gagatattcg acattggccg
     3121 tcaagaaaaa ccgcatctcg ccttttcgta tggtccacat agctgcatcg gcggtgagct
     3181 ggcgcgcctg gaactcaaag cggtctttgg ctcaatcttt cagcgctttc ccgcgcttcg
     3241 tctggcagtc gcaccagaac agctcaaact gcggaaagag atcattactg ggggcttcga
     3301 agaatttcca gtgttctggt aataaggatg agctctacaa ataagcagag gtggttgtgt
     3361 tgcgaaaaaa aaaaaaaaac accctaacgg gtgttttttt ttttttggtg tccccacgtg
     3421 tggcgctgga gaccgtccaa tggcggcgcg ccatcgaatg gtgcaaaacc tttcgcggta
     3481 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgagc ccgaaacgtc
     3541 gtacccaggc agaacgtgca atggaaaccc agggtaaact gattgcagca gcactgggtg
     3601 ttctgcgtga aaaaggttat gcaggttttc gtattgcaga tgttccgggt gcagccggtg
     3661 ttagccgtgg tgcacagagc catcattttc cgaccaaact ggaactgctg ctggcaacct
     3721 ttgaatggct gtatgagcag attaccgaac gtagccgtgc acgtctggca aaactgaaac
     3781 cggaagatga tgttattcag cagatgctgg atgatgcagc agattttttt ctggatgatg
     3841 attttagcat cggcctggat ctgattgttg cagcagatcg tgatccggca ctgcgtgaag
     3901 gtattctgcg taccgttgaa cgtaatcgtt ttgttgttga agatatgtgg ctgggtgtgc
     3961 tggtgagccg tggtctgagc cgtgatgatg ccgaagatat tctgtggctg atttttaaca
     4021 gcgttcgtgg tctgacagtt cgtagcctgt ggcagaaaga taaagaacgt tttgaacgtg
     4081 tgcgtaatag caccctggaa attgcacgtg aacgttatgc aaaattcaaa cgttgataag
     4141 gatcctaatt ggtaacgaat cagacaattg acggctcgag ggagtagcat agggtttgca
     4201 gaatccctgc ttcgtccatt tgacaggcac attatgcatc gatgataagc tgtcaaacat
     4261 gagcagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     4321 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4381 atgagcaaat attttatctg aggtgcttcc tcgctc
//
