LOCUS       sequence-423263-        4416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Micromonospora mirobrigensis LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4416)
  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 4416)
  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: pCYP40
FEATURES             Location/Qualifiers
     source          1..4416
                     /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      2490..2499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4328..4347)
                     /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 tgtttaaaat tttagcagga
     2101 ggaatttaaa atgagctcac agcaatcccc aatcccttat ccgttagacc cagcgccgac
     2161 gatataccat ccggcccccg catatactga attgcgggaa cgttgccctg tcgcccatgt
     2221 agccctgccg gatggcacca gcgcttacct ggctacccgc catgccgatg tgcgccgtgt
     2281 gttcactgac caacgtttca gccgtgccgc agcagctgga ccgaatcggc caaccagaga
     2341 actcggttcc ctcgcagagg atagtctcat tggtatggac cctccgcgcc acacaactat
     2401 gcgacgtgcc gtcagtcatg catttacggt acgccgtgtt gaagcgttgc gaccgacggt
     2461 cgccaccctg gttgacggta tgattgatgc catggtggcc gcgggaccgc ctgccgacct
     2521 tattacccac ttaagcgcac ccctgccgat ttacgtcatt tcaaaactgt ttggtatccc
     2581 ggaacaagat cgtgaacgtg tgcgtcagtg gtctgatgca ttggttggtg attgggatgc
     2641 cgatccggcg gctcccaagg cggcgctgga tgcattcgca gaactgattg cccagcgacg
     2701 tcgtgaacct ggggacgatc tgatgtcagc attgattggg gcatgggaca cgcacgatga
     2761 cctgaccgaa agagagctgg tttcggtgac cgctggcatc tttgttggtg gccatgaaac
     2821 aacgacgaac caacttaact tatttctgct ggtgctggca cgtcatccgg aacagctggc
     2881 aggcttgcgg ggtgatgacc cggtggcagt agcccgagct gtggaggaac tgtctcgttt
     2941 tatccagttg ggcgaccacg gcgtgttgtt accgagagtg accaccgaag aagtgcagct
     3001 gggcggagta cggttgccag cgggggcacc cgttttgccg gccattgcga gcgcaaatcg
     3061 tgatgaagaa gtattcacgg gcgcagatag actggatctc acccgtgcgc acaacccgca
     3121 tctggcgttc ggcgctgggc ctcaccactg tctgggcgca gcgttggcac gaatggaact
     3181 gcaagaggcc ctgggtgcgt tactgcgtag actgccgggt ctgcgcctgg ccgttgacga
     3241 agcgcgcctt cgctttcgtc ccgggcttgt agtacgctcc ctggaagcct taccggtgac
     3301 gtgggatacc gtggaccgat aataaggatg agctctacaa ataagcagag gtggttgtgt
     3361 tgcgaaaaaa aaaaaaaaac accctaacgg gtgttttttt ttttttggtg tccccacgtg
     3421 tggcgctgga gaccgtccaa tggcggcgcg ccatcgaatg gtgcaaaacc tttcgcggta
     3481 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgagc ccgaaacgtc
     3541 gtacccaggc agaacgtgca atggaaaccc agggtaaact gattgcagca gcactgggtg
     3601 ttctgcgtga aaaaggttat gcaggttttc gtattgcaga tgttccgggt gcagccggtg
     3661 ttagccgtgg tgcacagagc catcattttc cgaccaaact ggaactgctg ctggcaacct
     3721 ttgaatggct gtatgagcag attaccgaac gtagccgtgc acgtctggca aaactgaaac
     3781 cggaagatga tgttattcag cagatgctgg atgatgcagc agattttttt ctggatgatg
     3841 attttagcat cggcctggat ctgattgttg cagcagatcg tgatccggca ctgcgtgaag
     3901 gtattctgcg taccgttgaa cgtaatcgtt ttgttgttga agatatgtgg ctgggtgtgc
     3961 tggtgagccg tggtctgagc cgtgatgatg ccgaagatat tctgtggctg atttttaaca
     4021 gcgttcgtgg tctgacagtt cgtagcctgt ggcagaaaga taaagaacgt tttgaacgtg
     4081 tgcgtaatag caccctggaa attgcacgtg aacgttatgc aaaattcaaa cgttgataag
     4141 gatcctaatt ggtaacgaat cagacaattg acggctcgag ggagtagcat agggtttgca
     4201 gaatccctgc ttcgtccatt tgacaggcac attatgcatc gatgataagc tgtcaaacat
     4261 gagcagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     4321 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4381 atgagcaaat attttatctg aggtgcttcc tcgctc
//
