LOCUS       sequence-423536-        4584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Nonomuraea jiangxiensis LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4584)
  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 4584)
  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: pCYP21
FEATURES             Location/Qualifiers
     source          1..4584
                     /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(4496..4515)
                     /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 tgtttaggat atagtagcac
     2101 gaagggggta cttaatgttt aacgccactg gcacgaccgt aacacgcccc aaagttatcc
     2161 cattctacag agccattcca cgtttggtta aagatccagt gaccgagctt gctcgtatgg
     2221 gaacggaagc agggggacgt gtggtgcggt tagaccttgg tccgtttcgt ccgtatctga
     2281 tcagccaccc ggaccatgtg caacaggtac tgaaaacgga ttgggataat tttgtccgtg
     2341 cgggcatgtt ctggcaacct ctgcagcgcc tcatcgggca ctctataatg ggagaaggcg
     2401 agtcgtgggc aagtgcgcgc aaaatcctcc agcccctgtt tacggcacgc tacaccgcgt
     2461 ccctggctga agatatggcg gatatcatag ccgagcgcat tgtggaactg gatgattttg
     2521 cgcaggcagg ccgcccgatc gatgttgaac atgaaatgac gtctattgtt aatcaggcgg
     2581 ttattaaagt tctgttcggt ggtcgcataa ctcgagaaac agcagaacgc ttagcacccg
     2641 agttcgccac atgcgccagt tctatagggt tccgtatgct gttcccgtcg gctccttact
     2701 cgatccgcgt accaggagac agagccttcc tgcgtgccac gaagaaaatc catgagatcg
     2761 tttatcccct gatcgagcgt gctcagcgtg aaaagacgca gagcaaggaa gtggtgtcag
     2821 ccttactgac ggtgcgcctg gaacaggaag gtaaggccga cgttcgccaa gttcgtgatg
     2881 acttagtgaa tgtatatggt gcggcaagcg agaataccgc gatgtccctg ctgtggctgt
     2941 ggagtgtcct gcacgaccat ccggaagtaa acgcaagatt gcaggaggag attcaccgag
     3001 ttgtgggtga tggtccggtc cgtccttccc acgtcccgga aatgcattac ctgcgcatgg
     3061 tgcttcaaga actgctgcgc gtgtatcctt ctgcttgggt tttacctcgc caagcggtag
     3121 cggatacgga aattggtggt gtccgcatta aagccggaag ccaggtactg gttagcccct
     3181 acactacaca tagactcgaa gaattttggc ctcgcccgac cgaatttgat ccagagagat
     3241 gggccgggga taagacggaa agacgccatc gttactcctt tatcccgttt ggcgctggtc
     3301 cgcatgtctg cattggccag catttattct acgtggaggc ctcgctgttg attgccaatc
     3361 ttttctctcg ctaccgcccg gcgctgccgt acccgcagaa acttacgcca cttcgtggcg
     3421 gggcgacgct ccgcccgaaa gaaaagctgt ttttgcaact gcatccgatc gaacgcaagc
     3481 gcgccgcgta ataaggatga gctctacaaa taagcagagg tggttgtgtt gcgaaaaaaa
     3541 aaaaaaaaca ccctaacggg tgtttttttt ttttggtgtc cccacgtgtg gcgctggaga
     3601 ccgtccaatg gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     3661 gcccggaaga gagtcaattc agggtggtga atatgagccc gaaacgtcgt acccaggcag
     3721 aacgtgcaat ggaaacccag ggtaaactga ttgcagcagc actgggtgtt ctgcgtgaaa
     3781 aaggttatgc aggttttcgt attgcagatg ttccgggtgc agccggtgtt agccgtggtg
     3841 cacagagcca tcattttccg accaaactgg aactgctgct ggcaaccttt gaatggctgt
     3901 atgagcagat taccgaacgt agccgtgcac gtctggcaaa actgaaaccg gaagatgatg
     3961 ttattcagca gatgctggat gatgcagcag atttttttct ggatgatgat tttagcatcg
     4021 gcctggatct gattgttgca gcagatcgtg atccggcact gcgtgaaggt attctgcgta
     4081 ccgttgaacg taatcgtttt gttgttgaag atatgtggct gggtgtgctg gtgagccgtg
     4141 gtctgagccg tgatgatgcc gaagatattc tgtggctgat ttttaacagc gttcgtggtc
     4201 tgacagttcg tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg cgtaatagca
     4261 ccctggaaat tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga tcctaattgg
     4321 taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga atccctgctt
     4381 cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga gcagatcctc
     4441 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     4501 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcaaatat
     4561 tttatctgag gtgcttcctc gctc
//
