LOCUS       sequence-423534-        4661 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the Micromonospora cremea LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4661)
  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 4661)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pCYP15
FEATURES             Location/Qualifiers
     source          1..4661
                     /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(4573..4592)
                     /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 tgtttagaat cgtacaggga
     2101 ggatttatat gttgcacgaa tatctgaccc cgcattcgcg ctcagcaacc ccgtcaatcc
     2161 ataagggtgt aaccgtgccg gtggcagcgc cacccaccgt tcctgtccaa cctcgccgtg
     2221 caggctccgt ccccctgcgt catgtcgtgc cgaacttatt tcgagatcct gcgcgaacct
     2281 tggtggagtt gggggaacgg tcgcagggtg aggtcgtgaa attgtctttg ggcgccttcc
     2341 gtccgtattt aatcacccgg cctgaacatc tgcaacgcgt gcttcgtgaa aagtccgata
     2401 actatgtgcg tgccggcgat gggttgcaat ggcgaccgct taaacgttta tttggcgatg
     2461 ggattctgag tgacggcgat tattggaaaa atagtcgcca cgtgctgcag ccattgttca
     2521 ccgcgcgccg ggtggattct ctggtcgatc gcctggctga agctattgag gaggtcgtag
     2581 acgaacttga tgaaccggcg cgggccggtc agccggttga catgggaacg gtgcaagcgc
     2641 gtattgtatg ctccgccatt atgcgtgtgt tcttcgcgga caaaatctca gtgcctcagg
     2701 cgatgagaat tatggaagcg caagatgcca tcgcgtggag cgttatgcca cgtctgttag
     2761 tgccgtgggc tccgttggcg atgccaatgc cgggggatcg cgcctttcgc caggccgttc
     2821 aactcattga tgacactgtt cttccgattg tgcgcgccgc tcgtggcaca gccgaaacgg
     2881 atggcgatga tatcatagca accttgtggc gcggtcgtac ggaagccggt ggtcgcctgg
     2941 acgaacgcca agtgcgcgat gataccgtat caatggttgc tacgacgacg gaaacgacca
     3001 ttagcgtatt aacctggctt tggccgcaca tcgctcaaga tgaacaaatc gcggaacgcc
     3061 tgtacgagga gattgatcgt gttgttggtg acgcaccggt ccgccgtgag cacttgcgcg
     3121 aactgcgtta tacccgacag gtgctggatg aactgcttcg cctgtatccg attggctggc
     3181 tctttcctcg tcgggcgtta gaggcagacg tcatcggggg cgttcgcatc gaagcaggcg
     3241 ctaccctggt tgtgtcaccg ctgatcaccc accgtatgag tgcgttttgg gatcgccctg
     3301 aagtgtttga tcctgatcgt tttgaccctg aacaggtgag aaatcgtcat cgctatgcgc
     3361 actttccgtt cggtggcggt ccgcaccagt gtatcggcat gcatctgttc tacctggagg
     3421 ccatgttgat agtcgctacc ttactgtctc gctttagatt tcgcctggag cggcaggctc
     3481 tgcccgggat aaaagtggcg gcagctttgc gccctctgca gcgggtcgag gtcacactgc
     3541 agccggttcg gagtgtctcg gcctaataag gatgagctct acaaataagc agaggtggtt
     3601 gtgttgcgaa aaaaaaaaaa aaacacccta acgggtgttt tttttttttt tggtgtcccc
     3661 acgtgtggcg ctggagaccg tccaatggcg gcgcgccatc gaatggtgca aaacctttcg
     3721 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaata tgagcccgaa
     3781 acgtcgtacc caggcagaac gtgcaatgga aacccagggt aaactgattg cagcagcact
     3841 gggtgttctg cgtgaaaaag gttatgcagg ttttcgtatt gcagatgttc cgggtgcagc
     3901 cggtgttagc cgtggtgcac agagccatca ttttccgacc aaactggaac tgctgctggc
     3961 aacctttgaa tggctgtatg agcagattac cgaacgtagc cgtgcacgtc tggcaaaact
     4021 gaaaccggaa gatgatgtta ttcagcagat gctggatgat gcagcagatt tttttctgga
     4081 tgatgatttt agcatcggcc tggatctgat tgttgcagca gatcgtgatc cggcactgcg
     4141 tgaaggtatt ctgcgtaccg ttgaacgtaa tcgttttgtt gttgaagata tgtggctggg
     4201 tgtgctggtg agccgtggtc tgagccgtga tgatgccgaa gatattctgt ggctgatttt
     4261 taacagcgtt cgtggtctga cagttcgtag cctgtggcag aaagataaag aacgttttga
     4321 acgtgtgcgt aatagcaccc tggaaattgc acgtgaacgt tatgcaaaat tcaaacgttg
     4381 ataaggatcc taattggtaa cgaatcagac aattgacggc tcgagggagt agcatagggt
     4441 ttgcagaatc cctgcttcgt ccatttgaca ggcacattat gcatcgatga taagctgtca
     4501 aacatgagca gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     4561 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     4621 ggctcatgag caaatatttt atctgaggtg cttcctcgct c
//
