LOCUS       sequence-424412-        4425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces griseolus LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4425)
  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 4425)
  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: pCYP58
FEATURES             Location/Qualifiers
     source          1..4425
                     /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(4337..4356)
                     /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 tgtttagagc aacatatccc
     2101 taggagaatt aaaatgacgg ataccgccac cacaccccag accacagatg ccccggcctt
     2161 ccctagcaac cgttcctgcc cgtaccaact cccggatggc tatgcacaac tccgtgatac
     2221 accgggcccg ttacaccgcg tgacattgta cgatggccgt caagcgtggg tggtgaccaa
     2281 gcatgaagcc gcacgtaaac tgcttggcga cccacgttta tcgtcaaatc gtacggacga
     2341 caatttcccg gcaacatccc cgcgttttga agcagtgcgt gagagcccac aggcctttat
     2401 cggcctggac ccgcccgaac atggtacgcg ccgtcggatg acgattagtg agttcacggt
     2461 taaacgcatt aagggcatgc gtccggaagt cgaagaggtg gttcacggtt tcctggatga
     2521 aatgctggcc gctggcccaa ctgcagactt agtcagtcaa tttgcgttgc cggttccatc
     2581 gatggtgatt tgccgcctct taggcgttcc gtatgcggac catgaattct tccaggatgc
     2641 gagtaaacgc ttggtgcaaa gtaccgacgc acagtccgcc ttgaccgccc gcaacgatct
     2701 ggccggttat ttggacggac tgattaccca gtttcaaacg gagccaggcg cgggattggt
     2761 tggcgccctt gtcgctgacc agctggcaaa tggtgaaatt gaccgagagg aattaatcag
     2821 caccgccatg ctgctgctga tcgcggggca cgaaaccacc gcctctatga cgtcattatc
     2881 ggtaattacg ctgctggatc atcctgaaca gtacgcggcc ctgcgtgcgg atcgttcgct
     2941 cgttcctggt gcagtggagg aacttctccg ctacttagcg attgccgaca ttgccggcgg
     3001 ccgtgttgcg acagccgata tcgaagtgga gggacatctg atacgtgcgg gtgaaggggt
     3061 gattgtggtg aacagcattg cgaaccgcga tggcaccgtt tatgaagatc cggatgcgct
     3121 ggatatacat cgtagtgcac ggcatcactt agcgttcggt tttggcgttc accagtgctt
     3181 agggcagaat ttggcccgct tagagttgga ggtcatcctt aacgccctga tggatcgggt
     3241 accgaccctg cgcttagccg ttcctgttga acagctggta ctccgtccgg gcaccaccat
     3301 tcaaggtgtt aacgaattgc cggtcacttg gtgaggatga gctctacaaa taagcagagg
     3361 tggttgtgtt gcgaaaaaaa aaaaaaaaca ccctaacggg tgtttttttt tttttggtgt
     3421 ccccacgtgt ggcgctggag accgtccaat ggcggcgcgc catcgaatgg tgcaaaacct
     3481 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgagcc
     3541 cgaaacgtcg tacccaggca gaacgtgcaa tggaaaccca gggtaaactg attgcagcag
     3601 cactgggtgt tctgcgtgaa aaaggttatg caggttttcg tattgcagat gttccgggtg
     3661 cagccggtgt tagccgtggt gcacagagcc atcattttcc gaccaaactg gaactgctgc
     3721 tggcaacctt tgaatggctg tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa
     3781 aactgaaacc ggaagatgat gttattcagc agatgctgga tgatgcagca gatttttttc
     3841 tggatgatga ttttagcatc ggcctggatc tgattgttgc agcagatcgt gatccggcac
     3901 tgcgtgaagg tattctgcgt accgttgaac gtaatcgttt tgttgttgaa gatatgtggc
     3961 tgggtgtgct ggtgagccgt ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga
     4021 tttttaacag cgttcgtggt ctgacagttc gtagcctgtg gcagaaagat aaagaacgtt
     4081 ttgaacgtgt gcgtaatagc accctggaaa ttgcacgtga acgttatgca aaattcaaac
     4141 gttgataagg atcctaattg gtaacgaatc agacaattga cggctcgagg gagtagcata
     4201 gggtttgcag aatccctgct tcgtccattt gacaggcaca ttatgcatcg atgataagct
     4261 gtcaaacatg agcagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     4321 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     4381 ttcgggctca tgagcaaata ttttatctga ggtgcttcct cgctc
//
