LOCUS       sequence-423268-        4449 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the Streptomyces cyslabdanicus LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4449)
  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 4449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCYP46
FEATURES             Location/Qualifiers
     source          1..4449
                     /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(4361..4380)
                     /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 tgtttacggc tatttttaaa
     2101 aaataaggag ataaattatg actgcagccc cgaatgctgt tccactgtat ggcccagaac
     2161 atgccgctga cccgcaacgc tcctaccaat tactgcgtga gcagggtccg gttggccgag
     2221 cagagatcga tccgggatta gtggtctggg tcgtcactga ctatcgtgcg gcgttggatc
     2281 tcctgcaaga tcccggtacc tggaccaaag acacccgtgg ctgggacacc accgtgcccg
     2341 ccgattcccc tgttcgtgag cccttgcagt ggcgtccatc actcgccttt gccgatggtg
     2401 aggaacatgc ccatctgcgt aaagtgatca ccgactcctt tgcgttatta gacccacaat
     2461 atttaagaga tgtcaccctc cgacacgcgg acaccttact ggacgaattt gcccatactg
     2521 gtcgtgccga cctggtagcg cagtatgcgc gtcgtctgcc cctgatggtt ctgaacaccc
     2581 tgtttggcat gccagatgaa ggtgcggacc agttaatgtc tgcgattgaa gccttaatgt
     2641 cgaccgtgcc gcaggcaaag gaagctggtg agcaggctct gaccgcgtac ctcggttcgc
     2701 tctatgcaat gaaaagcgcc cggcgcggtc gcgatctgac gtcttggttc atcgaccatc
     2761 ctcatggcct gggccctgaa gaggttatta accaggtact gatcacccag ggagcggctt
     2821 atggaactct ggcgggtctg attgcgaaca ctttagcaca gatcctgtcc gatcaacggt
     2881 accgtggcac gctgaccacg ggttccttgc cgattcgcca tgcgattgat gaggttctgt
     2941 gggcagatcc gccgctggca atgtattccc ttcaccgcgc ccggcgctcc gttactttcc
     3001 atcgcagagc gattgaggct ggcgaacctg tgatggtatc ctatgcagct gtgaatactt
     3061 gcccgcacac cggacaggtt gccgatgagc aacgctccga tcacaaagcg cacctggcat
     3121 tcgctgcagg cccacatagc tgtccagcag aggacattgt gttcgtggtt gctgtggccg
     3181 cgattgaacg ccttatgggg cgactgccgg atgtggaact caccgttccg gccgcgaccc
     3241 tgacctatca gcccggtccg atttaccggg tgctgacctc tctgccctgc cgcttcacac
     3301 cgcagacacc ggactcacgt cgaagcacca gaggttccgt tcctgtttct gcgtaataag
     3361 gatgagctct acaaataagc agaggtggtt gtgttgcgaa aaaaaaaaaa aacaccctaa
     3421 cgggtgtttt tttttttttg gtgtccccac gtgtggcgct ggagaccgtc caatggcggc
     3481 gcgccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     3541 aattcagggt ggtgaatatg agcccgaaac gtcgtaccca ggcagaacgt gcaatggaaa
     3601 cccagggtaa actgattgca gcagcactgg gtgttctgcg tgaaaaaggt tatgcaggtt
     3661 ttcgtattgc agatgttccg ggtgcagccg gtgttagccg tggtgcacag agccatcatt
     3721 ttccgaccaa actggaactg ctgctggcaa cctttgaatg gctgtatgag cagattaccg
     3781 aacgtagccg tgcacgtctg gcaaaactga aaccggaaga tgatgttatt cagcagatgc
     3841 tggatgatgc agcagatttt tttctggatg atgattttag catcggcctg gatctgattg
     3901 ttgcagcaga tcgtgatccg gcactgcgtg aaggtattct gcgtaccgtt gaacgtaatc
     3961 gttttgttgt tgaagatatg tggctgggtg tgctggtgag ccgtggtctg agccgtgatg
     4021 atgccgaaga tattctgtgg ctgattttta acagcgttcg tggtctgaca gttcgtagcc
     4081 tgtggcagaa agataaagaa cgttttgaac gtgtgcgtaa tagcaccctg gaaattgcac
     4141 gtgaacgtta tgcaaaattc aaacgttgat aaggatccta attggtaacg aatcagacaa
     4201 ttgacggctc gagggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg
     4261 cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc cggacgcatc
     4321 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     4381 gatggggaag atcgggctcg ccacttcggg ctcatgagca aatattttat ctgaggtgct
     4441 tcctcgctc
//
