LOCUS       sequence-423282-        4600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sphaerisporangium album LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4600)
  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 4600)
  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: pCYP19
FEATURES             Location/Qualifiers
     source          1..4600
                     /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(4512..4531)
                     /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 tgtttacacg agggatctac
     2101 aaaataagga gaaagttatg gaaaacgttg cgatgccggt cgaaattaat gcagctcgta
     2161 cgggaaaacg cgttctcgcc cgtgttgcgc cagcgatcgt gcgtgatccg gttaacggcc
     2221 tcgcagattt aggccgccgc gcgggtggtc gtatcgttcg cttaaatctt ggtgtgctgc
     2281 gtccgtacct ggtcacgcac ccggaccacg tccagcatgt tatgcgcgat aattggacta
     2341 attactctcg tcaaggcatg ttctggcagc ccttagggcg tctgctgggc gattcgattc
     2401 tcggtgatgg cccaggttgg gaggcaagcc gccgtatttt gcagcctctg tttaccccga
     2461 agtacgtcgc agcgctggcg ggcacaatcg ccgcaactgt tgatgctcgc atcgaagaac
     2521 tgaatgcacg cgccagatcc ggcgacccta tggacgccgg tcgcaccatg ggtgctatcg
     2581 ttaaccagtc tgttgttcgc gtgctgtttg gcgatcgcat tagtctgcgg gacggagaac
     2641 gtctgacgag cgcgtaccac accgcggacg cggcggtgtt ctctcgactc ctgctgccgt
     2701 ttatgccgta ccgtgttccg atgccggggg atcgtgcttt taaaacggcg gttgcgacca
     2761 tcgatgaaat catcttcccg ctgatccgcc aggtgcgccg tgaacctgac gacggctcgg
     2821 atgtggtaag tgccctctgt cgctcacgcg aagccgatgg cactccccgg gaggaccgtc
     2881 gtgtacgtga cgatttggtc agcatttacg cggcagcaag cgtgactacc gcggcaacct
     2941 taacctggac atggccgctt ctggaagcac atccagaagt agctgcaaaa gtttatgcgg
     3001 aaattgatca agtcgtaggt gatggtccgg cccttccagc gcacgttcca gaacttcgtt
     3061 atacgaaaat ggtgctccag gaggttatgc gactgtatcc cagtggttgg ttactgccac
     3121 gtaccgtagt cgaatccgaa acactgggcg gcacccgcct gaaagcgggc tccatggttc
     3181 tgatctcgcc ttatgtcacc catcgtctgg aagaggtgtg ggacagaccg cttgaatttg
     3241 acccagagcg tttcagtcct gaccgtgggg agcgtcgcca tcgttacgcg tattttccct
     3301 ttggtggagg cgcgcatcag tgtgttggaa aacatctgtt ctatatggat gctcccttaa
     3361 ttatggcagc cattctgagt cgttttcgac ccgcagctct gggtggacgt acgccagccc
     3421 cagctgctac ggcgctgtta cgtccacgtg gtcgtgtcgg tttacgactg ttgccagcta
     3481 aatgccctgg tccgggcacc cgttaataag gatgagctct acaaataagc agaggtggtt
     3541 gtgttgcgaa aaaaaaaaaa aaacacccta acgggtgttt tttttttttt ggtgtcccca
     3601 cgtgtggcgc tggagaccgt ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc
     3661 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatat gagcccgaaa
     3721 cgtcgtaccc aggcagaacg tgcaatggaa acccagggta aactgattgc agcagcactg
     3781 ggtgttctgc gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc
     3841 ggtgttagcc gtggtgcaca gagccatcat tttccgacca aactggaact gctgctggca
     3901 acctttgaat ggctgtatga gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg
     3961 aaaccggaag atgatgttat tcagcagatg ctggatgatg cagcagattt ttttctggat
     4021 gatgatttta gcatcggcct ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt
     4081 gaaggtattc tgcgtaccgt tgaacgtaat cgttttgttg ttgaagatat gtggctgggt
     4141 gtgctggtga gccgtggtct gagccgtgat gatgccgaag atattctgtg gctgattttt
     4201 aacagcgttc gtggtctgac agttcgtagc ctgtggcaga aagataaaga acgttttgaa
     4261 cgtgtgcgta atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga
     4321 taaggatcct aattggtaac gaatcagaca attgacggct cgagggagta gcatagggtt
     4381 tgcagaatcc ctgcttcgtc catttgacag gcacattatg catcgatgat aagctgtcaa
     4441 acatgagcag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4501 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4561 gctcatgagc aaatatttta tctgaggtgc ttcctcgctc
//
