LOCUS       sequence-423562-        4571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces clavuligerus LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4571)
  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 4571)
  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: pCYP29
FEATURES             Location/Qualifiers
     source          1..4571
                     /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(4483..4502)
                     /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 tgtttaggcc ccccatataa
     2101 aattggagta tttttatgac tacacagaaa cacgattcgt ctacaaccgg gaatttcacc
     2161 actggccgcg ccccaggagc gctcccatta atcgggcacg caggccatct gatgcgccgt
     2221 cctatcacat ttctggaaag ccttagcacg tacggagatt tagtcgagat tcgtatcggt
     2281 acccaacgcg cgtatgtccc atgtcacccg gaactgttac gccaggttct cactgacgat
     2341 cggaccttcg ataaaggtgg agtaattatc gagcgttttc gttccgtgct gggaaacggc
     2401 ttgggaacat gtccacactc cgaccaccgt cggcagcgcc gtctgatgca gccggcattc
     2461 tcaaaagcgc gcctggaaga gtatagtgct gtgatgcggc aagaaatcac cgcgctctct
     2521 ggctcgtggc aagagggaca agagattgac gcctttgccg tgttgtacgg tctgggcctg
     2581 cgagtcgtag tgcgcacact gctgagctcc gatctggatg atccgaccgt tgaacaagtc
     2641 agacatagtt ttgagcgcac gttacacgag ttcctgccgc gcacaattat cccgcagcaa
     2701 atccagcaac tgccattagc cgtgaataga agataccacc aggaattgca agcgctggac
     2761 gaggtcgtta accgcatcat tgctgactgt cgtgacgctg gtttggaaag aggcggtctg
     2821 ctggccaccc tgacggcgag ccatgcagga ggcggagttc ctatggatga tagtgagatt
     2881 cacgatcagg tgatcaccat gctgttaggc ggatcggaaa gcaccggcac gaccctgact
     2941 tgggccctgt atctgctgag tcagcatccc gaaattgaaa tgcgtctgca tgaagaggtg
     3001 gataccgtcc tcaacggacg tccagcagat tggaacgatt tgccaaatct ctcacttacc
     3061 cacaatatca ttacggaaac gttgcgtctg tatccgccag gttggctgct gagccggacc
     3121 acaacaaaag aagttgaact ggccggaacc cggatggcac cggatacgac tattgtattt
     3181 tcgccccacg taatccaacg acagggtgat attcatggat gcccagatgt tttcgacccg
     3241 gaccgttggt cgcccgatag aacggcggac ctgccgcgtg gtaattttgt gagttttggc
     3301 ggcgggcccc gtaaatgtat tggtgacact tttggtgaag ttcagagcac cctggcactg
     3361 agtacgattg cagcgcgctg gaaaatggaa tgtgccccag ggagtgatat tcgtccggtt
     3421 gcctttgact ttgcatatcg tccgcgccgg cttatcctgc gccctcgcct ccgataataa
     3481 ggatgagctc tacaaataag cagaggtggt tgtgttgcga aaaaaaaaaa aaaaacaccc
     3541 taacgggtgt tttttttttt tggtgtcccc acgtgtggcg ctggagaccg tccaatggcg
     3601 gcgcgccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     3661 tcaattcagg gtggtgaata tgagcccgaa acgtcgtacc caggcagaac gtgcaatgga
     3721 aacccagggt aaactgattg cagcagcact gggtgttctg cgtgaaaaag gttatgcagg
     3781 ttttcgtatt gcagatgttc cgggtgcagc cggtgttagc cgtggtgcac agagccatca
     3841 ttttccgacc aaactggaac tgctgctggc aacctttgaa tggctgtatg agcagattac
     3901 cgaacgtagc cgtgcacgtc tggcaaaact gaaaccggaa gatgatgtta ttcagcagat
     3961 gctggatgat gcagcagatt tttttctgga tgatgatttt agcatcggcc tggatctgat
     4021 tgttgcagca gatcgtgatc cggcactgcg tgaaggtatt ctgcgtaccg ttgaacgtaa
     4081 tcgttttgtt gttgaagata tgtggctggg tgtgctggtg agccgtggtc tgagccgtga
     4141 tgatgccgaa gatattctgt ggctgatttt taacagcgtt cgtggtctga cagttcgtag
     4201 cctgtggcag aaagataaag aacgttttga acgtgtgcgt aatagcaccc tggaaattgc
     4261 acgtgaacgt tatgcaaaat tcaaacgttg ataaggatcc taattggtaa cgaatcagac
     4321 aattgacggc tcgagggagt agcatagggt ttgcagaatc cctgcttcgt ccatttgaca
     4381 ggcacattat gcatcgatga taagctgtca aacatgagca gatcctctac gccggacgca
     4441 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     4501 ccgatgggga agatcgggct cgccacttcg ggctcatgag caaatatttt atctgaggtg
     4561 cttcctcgct c
//
