LOCUS       sequence-420111-        4594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rhizobium aethiopicum LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4594)
  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 4594)
  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: pCYP01
FEATURES             Location/Qualifiers
     source          1..4594
                     /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             2097..2105
                     /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(4506..4525)
                     /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 tgtttagtaa aaaagttaag
     2101 gaggttaatt tcatggatat gctgctgaat ccgttaaacc gctggcgtcg tttacgtgat
     2161 gatatcccgg ttatgccggg cgcctttccg ttagtcggtc atctgcctgc gatcgtgtgt
     2221 gacctgccgc gtttacttcg ccgtgctgag agaactttag gctcgcattt ctggctggat
     2281 ttcggcccgg caggtcacct gattacgtgc ttagatccgg atgctttggc ccttttaaga
     2341 cacaaagaag tgtcatcagc cctgattgaa gagatggcgc cggatatttt gggtggaaca
     2401 ctggttacac tcgacggcag cgcacaccgc caggcgcggg atggtatcaa agcggcgttc
     2461 ctgccgcgcg gcttatcaga agctggcatt ggcgaacttt tcgaaccgat catccgcgca
     2521 caagttaaag cgtggcgcga tcgcggtgaa gtggccatcc tcccggacac tcgaaattta
     2581 atgctgaaac tgaccttttc cttgatgggc atccctgccc aagatctgtc agagtggcat
     2641 cgcaaatatc ggcagctgct gcagttaatg gtagccaccc ctattgatct tccaggcatg
     2701 cctcttagac gcggacgcgc cgcacgagat tggattgacg cgcagagccg acaatttatc
     2761 cgcgatgccc gcgccccggc cgcaagaacg ggcttaatca atgatatggt tagcgccttt
     2821 gattgctcgg atggtgccct ttccgatgat gtcctggtcg caaatatccg cctgttgtta
     2881 ctggcaggcc atgaaacctc ggcttcaaca atggcgtgga tggtcattga actggcccaa
     2941 cacccggagt tatgggatgc gctggtggaa gaagcacagc gtgtaggagc cgtgccgaca
     3001 ggccatgagg atttagctca gtgcccggtt gcggaagcac tgtttcgcga aacgctgcgt
     3061 atgcatcccg cgtctagtct ggttcctcgt cgtgccatgc aggaactgca gctgggcccg
     3121 cgtcgtatcc cggctggtac tcatctgtgt attccgcttc tgcacttctc tactagcccg
     3181 ttgttacacg aggcgccaga cccctttcgg ttgggccgct ggctgcagcg taccgaaccg
     3241 atccggccag tagatatgct ccaatttggc gcggggccgc acgtgtgtat gggttaccac
     3301 ctggtgtggt tagagctggt ccagtttagc atcgccctgg ccttaaccat gcaggaagcg
     3361 ggcgtgcgac cgcgtctgat gagcggtgta gaaaaaggac gccgttatta cccgaccgca
     3421 tccgtgcatg ataaccccca ccgtatcctc atgtcctggg accctttgcc gcgcgttaaa
     3481 cgtcagcgcc gtcgccgtta ataaggatga gctctacaaa taagcagagg tggttgtgtt
     3541 gcgaaaaaaa aaaaaaaaca ccctaacggg tgtttttttt ttttggtgtc cccacgtgtg
     3601 gcgctggaga ccgtccaatg gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg
     3661 gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgagccc gaaacgtcgt
     3721 acccaggcag aacgtgcaat ggaaacccag ggtaaactga ttgcagcagc actgggtgtt
     3781 ctgcgtgaaa aaggttatgc aggttttcgt attgcagatg ttccgggtgc agccggtgtt
     3841 agccgtggtg cacagagcca tcattttccg accaaactgg aactgctgct ggcaaccttt
     3901 gaatggctgt atgagcagat taccgaacgt agccgtgcac gtctggcaaa actgaaaccg
     3961 gaagatgatg ttattcagca gatgctggat gatgcagcag atttttttct ggatgatgat
     4021 tttagcatcg gcctggatct gattgttgca gcagatcgtg atccggcact gcgtgaaggt
     4081 attctgcgta ccgttgaacg taatcgtttt gttgttgaag atatgtggct gggtgtgctg
     4141 gtgagccgtg gtctgagccg tgatgatgcc gaagatattc tgtggctgat ttttaacagc
     4201 gttcgtggtc tgacagttcg tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg
     4261 cgtaatagca ccctggaaat tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga
     4321 tcctaattgg taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga
     4381 atccctgctt cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga
     4441 gcagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     4501 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     4561 gagcaaatat tttatctgag gtgcttcctc gctc
//
