LOCUS       sequence-423273-        4428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Micromonospora humi LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4428)
  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 4428)
  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: pCYP42
FEATURES             Location/Qualifiers
     source          1..4428
                     /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)"
     regulatory      3278..3287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4340..4359)
                     /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 tgtttagaac ctgaacgtaa
     2101 ggagggaatt atgctggcca tggatatcga tgggggcctg cccgtggttg acagctaccg
     2161 tgttcctgtg aattcgggtt tgcgaccata cccatttgag cccttcactg gggatttacc
     2221 gactgaattg ctggacatgg tggtaaccga accggtttct cgggtgcttc tgccggatgg
     2281 ccgcgctgcc tggctggtac tgtcgtatgc gcactgttgc actgttctgg cagatcctcg
     2341 atttagtcgc cttccattag gcgcgactac cacccctgga gcaggaggtc cccgcgaatt
     2401 gaatatggat gggcctgctc atgccgctgt tcgtcgtgtt gccggccgag cgttcacagc
     2461 tcgtcgcatc gaaacctttc gtcctcgggt acgccgcctg gtggatgaac ttgtggatgc
     2521 gatgctggct ggcccgcgtc cagcagattt agtggcaggt ctggtcgctc cgctgcctgt
     2581 gttcgttgtt tgcgatgtgc tgggagtgcc ggttgcggat cggccacggt tctatgattg
     2641 gatctcaggt ctgaatagtg ttacagcgta tggcagcgct ggcgcggccg atgctcaagc
     2701 ggcattacgt gcgtacctga ccggacagtt ggcccgtaaa cgttcggcgc cgggggacga
     2761 tttgttgtct gcctgggtgc tgggacagga tgcgcacgcg ctggtggatg cagaactggt
     2821 agaactggcg atgggtgtac tgcttggtgg gcttgagata aacgccacca gcgcaggcct
     2881 gcgagcgctg tttcagcatc ccgagcagtt ggccaaactc cgtgcggctc caggcaaact
     2941 cgcagcggct accgacgaga tcttgcgcta tacctcggtg tcctcgatgt tccgggttca
     3001 ggtggtacgg gaggacctga cgcttggtgg cgtcgcgatg cgtgcgggtg attgcgtgat
     3061 ggccattccg tgggccggta atcgggaccc gcgtcacttc ccggaaccta atcgttttga
     3121 cattgatcgc gcgccgaccg tcccgcactt aacctttgga ttcggtcctc atttttgctt
     3181 gggtaccgcc cttggccgca tgcaggttga gctttcactt ggcgcgttac tgtctcgcat
     3241 gccagaactt gcgccggcgg tgcctattga agagatacca tggcggcacg atcgcgtcaa
     3301 tggtggcatc gcggcgtttc cggttacttg gtaataagga tgagctctac aaataagcag
     3361 aggtggttgt gttgcgaaaa aaaaaaaaaa caccctaacg ggtgtttttt ttttttttgg
     3421 tgtccccacg tgtggcgctg gagaccgtcc aatggcggcg cgccatcgaa tggtgcaaaa
     3481 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatatga
     3541 gcccgaaacg tcgtacccag gcagaacgtg caatggaaac ccagggtaaa ctgattgcag
     3601 cagcactggg tgttctgcgt gaaaaaggtt atgcaggttt tcgtattgca gatgttccgg
     3661 gtgcagccgg tgttagccgt ggtgcacaga gccatcattt tccgaccaaa ctggaactgc
     3721 tgctggcaac ctttgaatgg ctgtatgagc agattaccga acgtagccgt gcacgtctgg
     3781 caaaactgaa accggaagat gatgttattc agcagatgct ggatgatgca gcagattttt
     3841 ttctggatga tgattttagc atcggcctgg atctgattgt tgcagcagat cgtgatccgg
     3901 cactgcgtga aggtattctg cgtaccgttg aacgtaatcg ttttgttgtt gaagatatgt
     3961 ggctgggtgt gctggtgagc cgtggtctga gccgtgatga tgccgaagat attctgtggc
     4021 tgatttttaa cagcgttcgt ggtctgacag ttcgtagcct gtggcagaaa gataaagaac
     4081 gttttgaacg tgtgcgtaat agcaccctgg aaattgcacg tgaacgttat gcaaaattca
     4141 aacgttgata aggatcctaa ttggtaacga atcagacaat tgacggctcg agggagtagc
     4201 atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca tcgatgataa
     4261 gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     4321 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     4381 cacttcgggc tcatgagcaa atattttatc tgaggtgctt cctcgctc
//
