LOCUS       sequence-422118-        4539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rhizobium grahamii LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  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 4539)
  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: pCYP04
FEATURES             Location/Qualifiers
     source          1..4539
                     /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(4451..4470)
                     /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 tgtttaggct tcccaaggag
     2101 gtaatttatg ctgctgaatc cattaaaccg tcgccggcgt ttacagtaca atatcccgat
     2161 catgcacgga gcatttccta ttgttggtca tttaccggcg attgtgagcg atctgccgcg
     2221 tctgctgcgt cgtgcggagg gtatgttagg aaatcacttt tggttggatt tcggccccgg
     2281 gggccagttg atgatatgtc ttgacccaga tacattggca gcgctgcgcc ataagaatgt
     2341 cagctcatcg ttaattggcg aaatggcgcc agaaattctg ggcggcacac tggtgtcttt
     2401 agatggttcc gctcaccgtc gcgcccgcga cggaattaaa gaagcctttc tcccgaaagg
     2461 actgaccgcg gccggtatag gcgacctttt cgcaccgatc attcgcgccc gggtacaggc
     2521 ttggcgcgaa cgtgggggag tttccatttt accggaaacc ggtgatttga tgctgaaact
     2581 gaccttttac cttatgggca tcccgccaca ggacttacct gtttggcatc gtaaatatcg
     2641 tcagttactg cagcttatcg tggcccctcc agtggactta cctggtacac cgcttcgtcg
     2701 aggacgtgcc gcccgcgact ggatcgatgc tcagtctcgc caatttattc gcgacgcacg
     2761 cgcccggtct actcgtaccg ggttaatcaa tgatatggtg agtgcgtttg accgatctga
     2821 ggacgcgtta tcagatgacg tcttagtggc taacgtgcgc ctgttgttac tcgcgggtca
     2881 tgaaactaca gcctcaacga tggcgtggat ggttatcgaa cttgcacggc accctgacct
     2941 ctggcatgcc ctggtggagg aagcccgtca tgccggtact gtgccaaccc agcatgagca
     3001 cctcccgcag ttcccagttg cggaagcatt gttccgtgaa acgcttcgtt tacacccagc
     3061 tagtagcctg gtccctcgcc gcacaactcg tgagctgcac ctgggaggcc ggagactgcc
     3121 ggccggaact cgtctgggaa ttccgctgtt acatttctct acgagcagtc tgctgcacga
     3181 cgcaccggat cagttccgaa tagaacggtg gctgggccgc gttgagcccg tgggtccgat
     3241 cgatatgttg cagtttggga cggggccgca tgtttgtatt ggttaccatc tggtttggct
     3301 tgagctggtg cagttcgcag tcgcactggc gcttacgatg caggaatctg gggtcaaacc
     3361 gcagctctta aacgatgctg aaaaagggcg tcgctttttc ccgacagcac agcccagtat
     3421 gaaagtccgc attggatttt cctaataagg atgagctcta caaataagca gaggtggttg
     3481 tgttgcgaaa aaaaaaaaaa aacaccctaa cgggtgtttt tttttttttg gtgtccccac
     3541 gtgtggcgct ggagaccgtc caatggcggc gcgccatcga atggtgcaaa acctttcgcg
     3601 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatatg agcccgaaac
     3661 gtcgtaccca ggcagaacgt gcaatggaaa cccagggtaa actgattgca gcagcactgg
     3721 gtgttctgcg tgaaaaaggt tatgcaggtt ttcgtattgc agatgttccg ggtgcagccg
     3781 gtgttagccg tggtgcacag agccatcatt ttccgaccaa actggaactg ctgctggcaa
     3841 cctttgaatg gctgtatgag cagattaccg aacgtagccg tgcacgtctg gcaaaactga
     3901 aaccggaaga tgatgttatt cagcagatgc tggatgatgc agcagatttt tttctggatg
     3961 atgattttag catcggcctg gatctgattg ttgcagcaga tcgtgatccg gcactgcgtg
     4021 aaggtattct gcgtaccgtt gaacgtaatc gttttgttgt tgaagatatg tggctgggtg
     4081 tgctggtgag ccgtggtctg agccgtgatg atgccgaaga tattctgtgg ctgattttta
     4141 acagcgttcg tggtctgaca gttcgtagcc tgtggcagaa agataaagaa cgttttgaac
     4201 gtgtgcgtaa tagcaccctg gaaattgcac gtgaacgtta tgcaaaattc aaacgttgat
     4261 aaggatccta attggtaacg aatcagacaa ttgacggctc gagggagtag catagggttt
     4321 gcagaatccc tgcttcgtcc atttgacagg cacattatgc atcgatgata agctgtcaaa
     4381 catgagcaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     4441 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4501 ctcatgagca aatattttat ctgaggtgct tcctcgctc
//
