LOCUS       sequence-423392-        4410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sinorhizobium fredii LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4410)
  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 4410)
  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: pCYP14
FEATURES             Location/Qualifiers
     source          1..4410
                     /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)"
     RBS             2101..2109
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(4322..4341)
                     /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 tgtttacgct tctaaacccc
     2101 taaggaggtt gttatgccgg agcagccgtt gccgaccctt ccaatgtggc gcgtggatca
     2161 tatcgaaccg tcgcccacca tgcttgcgct gcgggcgaat ggtcctattc acaacgtgcg
     2221 atttccacgc gggcatgaag gttggtgggt cactggttat gacgaggcca aagccgtttt
     2281 gtcagatgcg gcgttccgtc cggcgggaat gccaccggca gcatttactc ccgattgcgt
     2341 gatcctgggt agccctgggt ggctggttag tcatgagggc ggtgagcacg cgcgtttaag
     2401 aaccattgtg gccccggcct ttagtgaccg tcgggttaaa ttactggctc aacaggttga
     2461 agcgattgcg gcgcaactgt ttgaaacttt agctgcacag ccgcagccag cggacttgcg
     2521 ccgtcacctg tcttttccgc tgccggccat ggttatttcg gctctgatgg gcgtgttgta
     2581 cgaggaccac gcattcttcg caggtctttc cgacgaggta atgacgcatc agcacgaatc
     2641 cggcccgcgc tcagcttctc gcttggcctg ggaggaattg cgtgcatata ttcgcggcaa
     2701 aatgcgcgac aaacgtcaag accctgggga taatctgctg accgacttac tggcagccgt
     2761 agatcgtggc gaggccaccg aagaggaggc aatcggactg gcggccggaa tgctcgtggc
     2821 tggccatgaa tcaacagtag ctcagatcga attcggcctg ctggcaatgc tgcgacatcc
     2881 gcagcaacgc gagcgcttgg tcggcaaccc ttcgctggta gacaaggcag ttgaagaaat
     2941 cttaagaatg tacccgcctg gggctggttg ggacggaata atgcgctatc cccgtaccga
     3001 tgtcactata gccggcgtcc acattccggc tgagtcgaaa gtgttagttg gactgcctgc
     3061 tacctccttt gacccacgtc actttgaaga tcctgaaatt tttgatatcg gccgtgacgc
     3121 taaaccgcac ttggccttct cctatggccc tcattattgt attggtatgg cgctggcacg
     3181 gttggaatta aaggtggttt ttggcagcat ctttcaacgc tttccggcac tgcgtcttgc
     3241 ggtcgcgcct gaggaactta agctgcgcaa ggaaattatt acaggtgggt ttgaagagtt
     3301 tccggttctg tggtaataag gatgagctct acaaataagc agaggtggtt gtgttgcgaa
     3361 aaaaaaaaaa aaacacccta acgggtgttt tttttttttt ggtgtcccca cgtgtggcgc
     3421 tggagaccgt ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc ggtatggcat
     3481 gatagcgccc ggaagagagt caattcaggg tggtgaatat gagcccgaaa cgtcgtaccc
     3541 aggcagaacg tgcaatggaa acccagggta aactgattgc agcagcactg ggtgttctgc
     3601 gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc ggtgttagcc
     3661 gtggtgcaca gagccatcat tttccgacca aactggaact gctgctggca acctttgaat
     3721 ggctgtatga gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg aaaccggaag
     3781 atgatgttat tcagcagatg ctggatgatg cagcagattt ttttctggat gatgatttta
     3841 gcatcggcct ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt gaaggtattc
     3901 tgcgtaccgt tgaacgtaat cgttttgttg ttgaagatat gtggctgggt gtgctggtga
     3961 gccgtggtct gagccgtgat gatgccgaag atattctgtg gctgattttt aacagcgttc
     4021 gtggtctgac agttcgtagc ctgtggcaga aagataaaga acgttttgaa cgtgtgcgta
     4081 atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga taaggatcct
     4141 aattggtaac gaatcagaca attgacggct cgagggagta gcatagggtt tgcagaatcc
     4201 ctgcttcgtc catttgacag gcacattatg catcgatgat aagctgtcaa acatgagcag
     4261 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     4321 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     4381 aaatatttta tctgaggtgc ttcctcgctc
//
