LOCUS       sequence-423537-        4616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces atratus LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4616)
  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 4616)
  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: pCYP23
FEATURES             Location/Qualifiers
     source          1..4616
                     /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(4528..4547)
                     /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 tgtttaggta cgtatcaagg
     2101 aggtaataca atgtatcgct cgcgatgcgg aaaagacggt cgtgttaccc cggacattga
     2161 tgtgaccgcc gatatggatg tgccgccgac agcacctggg cgcctgccgt taattgggca
     2221 cgtggtgcca ctcctgaaag atcgcctcga atttgttgaa cgccttcgta cttatgggcc
     2281 tattgtgaga atgagcgtag gtctgaaaaa ggtgaccgtg gtaaattcac atgaactgat
     2341 tcatgaaatg ctgacctcca agagtgggtc ctttagtaaa gggctgctcg gtgagaaact
     2401 gaaactgttt ggacagtcgg ctttgccagt cgcggaaggc gaaccacact tgcgccgtcg
     2461 ccgtctgctg cagccggcgt tccatcgtga acgcatttca gggtatgtgg atacgatgcg
     2521 agaaactgtc gaaccaagta tcacaggctg gcgcgagggt gaactgttag atttaagaac
     2581 cgtaatgggg ctcatggctc aggatgtggc catgtcagtg ctgtgtagct ttcgcgctga
     2641 acaagagcga gcgcgggcta tcctcgacgc agtggatacc gtgtttacag ctgcgatctg
     2701 gcggaccatg gtgccggttt cttccctgga gcgcctcccg accagaagca accagcgcct
     2761 taaagcagcc cgctctctgt tgcgttctga agttgcggat attatcgcgg aacgtcgtgc
     2821 tgcaccggat gcacacgacg atttggtaac cctgctttta gaagctcgcg atgaaacggg
     2881 ggccccgctg cctgacaacg agatcttatc agagattatc ggattgcttg ccgcgggaag
     2941 cgagactacc ggcgtcatgt tggcatggct gtttcatgag ttgggccgtg atccagagct
     3001 cgagcgtcgg ctgcacgaag aagttgatag tgttgttgga gaaggcccga ttactgctga
     3061 acacgtcccg cagttggctt tcacgcgtcg cctcgtgtcc gaaaccctgc gcctgcatca
     3121 cgctggctgg ctggttacac gtcaagcgac gaagcctgtg cggttaggcc aagtagttct
     3181 gcctgctggc gaacaagtcc tgtggagccc ctatgcattg caccgcgatc cagcattata
     3241 tccagatcct cttcgctttg atccggatag atggttgccg gaacgtccgc agccgccgcg
     3301 cggtgctttc atcccgtttg gttccggaaa acgcatgtgt attggtgacg cctttgcgtg
     3361 gactgaggcc atagtgatta ccgcctttat tgctagtcgt tggcgcctgc gcccggtgcc
     3421 tggtagcacg gtagagcctg tgggcctcct gacagtgcac cccagtgacc cccgtatggt
     3481 ggcggaaact cgtcaacgtg cagtggtggg taccgcggtg taataaggat gagctctaca
     3541 aataagcaga ggtggttgtg ttgcgaaaaa aaaaaaaaaa caccctaacg ggtgtttttt
     3601 ttttttggtg tccccacgtg tggcgctgga gaccgtccaa tggcggcgcg ccatcgaatg
     3661 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     3721 gaatatgagc ccgaaacgtc gtacccaggc agaacgtgca atggaaaccc agggtaaact
     3781 gattgcagca gcactgggtg ttctgcgtga aaaaggttat gcaggttttc gtattgcaga
     3841 tgttccgggt gcagccggtg ttagccgtgg tgcacagagc catcattttc cgaccaaact
     3901 ggaactgctg ctggcaacct ttgaatggct gtatgagcag attaccgaac gtagccgtgc
     3961 acgtctggca aaactgaaac cggaagatga tgttattcag cagatgctgg atgatgcagc
     4021 agattttttt ctggatgatg attttagcat cggcctggat ctgattgttg cagcagatcg
     4081 tgatccggca ctgcgtgaag gtattctgcg taccgttgaa cgtaatcgtt ttgttgttga
     4141 agatatgtgg ctgggtgtgc tggtgagccg tggtctgagc cgtgatgatg ccgaagatat
     4201 tctgtggctg atttttaaca gcgttcgtgg tctgacagtt cgtagcctgt ggcagaaaga
     4261 taaagaacgt tttgaacgtg tgcgtaatag caccctggaa attgcacgtg aacgttatgc
     4321 aaaattcaaa cgttgataag gatcctaatt ggtaacgaat cagacaattg acggctcgag
     4381 ggagtagcat agggtttgca gaatccctgc ttcgtccatt tgacaggcac attatgcatc
     4441 gatgataagc tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg gccggcatca
     4501 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     4561 gggctcgcca cttcgggctc atgagcaaat attttatctg aggtgcttcc tcgctc
//
