LOCUS       sequence-424494-        4396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces sp. Tu6071 LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4396)
  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 4396)
  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: pCYP48
FEATURES             Location/Qualifiers
     source          1..4396
                     /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             2098..2106
                     /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(4308..4327)
                     /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 tgtttaagga taaaacttaa
     2101 ggaggttact atggaccgtg ttctggactt tttgagctca gcagattcag ccggagaact
     2161 gcagcctcgt ctggttgaac tgtctcgaca ggagacactg ccgcgtgtgc tgctcgcgga
     2221 tggacaagaa gcgtggcttg tgacgcgtaa cgaagatgtg cgtaccgtat tatccgaccg
     2281 gagctttact cgtgatgtta tgggcgaacg cgcacgtcaa gccggcgaaa caccagacgg
     2341 tgcgcgtagt gtcaacatgg acggccgtcc ccataacgaa ctgcgcgcgc tggtgagcaa
     2401 agccttcacc gttcgccgca ttgaagccat gcgtccgcgg attcaggcct ggaccgatga
     2461 gctgatcgat gctatggaag aaaccggacc gccagccgac ctggtcgcac atctcgctgt
     2521 tccattacca gcgctggcaa tatgcgagct gttgggcttt ccggtcgaag atcgtcaggt
     2581 actgtcgggt tggtgcgaac ggatcaccag acttggagaa ggaggcccgg atcagcgggc
     2641 ctggcaggaa ctttcggcat atattgcgcg tcgtgtccca gtagagcgtg cagctgctcg
     2701 cggaggcctg gccccggaga catctatcct gacacgcctc gtacatgcgc atgatagcga
     2761 agatgccctt tccatggaag aactgctcag tctgaccgtt gttgtgctgg cgggcggctt
     2821 ggagacaacc caaacggcga tcggcgcggg gatggttcgt ttatttcgaa acccggccca
     2881 acttgataaa gtgcgtgcgg acccagatct ggtagtcccg gctgtcgagg agattctccg
     2941 ctatcagccg gtgattgatg ttaatagagt acaggtggcc acgcggacag tgcgactggg
     3001 cggacaggag attcgtgcgg gtgatcttgt ccaagtttct gttaacgccg caaatcgtga
     3061 cgaaaccgtt ttcccggatt ctgaacgctg cgacgtcacg cgcgggccga atccccacct
     3121 tgcattcggc tatggtgctc accattgtct gggtgctgcc ctggcacgtc tggaactgaa
     3181 aacggctttc tcgaccttgc tgcgtcgtct gccggatctg cgccccgcag ttccactgga
     3241 gtctttaggg tggcgtggtg gtcacgttac gctgggtctg gaggaactgc cagtcgcttg
     3301 gtaataagga tgagctctac aaataagcag aggtggttgt gttgcgaaaa aaaaaaaaac
     3361 accctaacgg gtgttttttt ttttttggtg tccccacgtg tggcgctgga gaccgtccaa
     3421 tggcggcgcg ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     3481 gagagtcaat tcagggtggt gaatatgagc ccgaaacgtc gtacccaggc agaacgtgca
     3541 atggaaaccc agggtaaact gattgcagca gcactgggtg ttctgcgtga aaaaggttat
     3601 gcaggttttc gtattgcaga tgttccgggt gcagccggtg ttagccgtgg tgcacagagc
     3661 catcattttc cgaccaaact ggaactgctg ctggcaacct ttgaatggct gtatgagcag
     3721 attaccgaac gtagccgtgc acgtctggca aaactgaaac cggaagatga tgttattcag
     3781 cagatgctgg atgatgcagc agattttttt ctggatgatg attttagcat cggcctggat
     3841 ctgattgttg cagcagatcg tgatccggca ctgcgtgaag gtattctgcg taccgttgaa
     3901 cgtaatcgtt ttgttgttga agatatgtgg ctgggtgtgc tggtgagccg tggtctgagc
     3961 cgtgatgatg ccgaagatat tctgtggctg atttttaaca gcgttcgtgg tctgacagtt
     4021 cgtagcctgt ggcagaaaga taaagaacgt tttgaacgtg tgcgtaatag caccctggaa
     4081 attgcacgtg aacgttatgc aaaattcaaa cgttgataag gatcctaatt ggtaacgaat
     4141 cagacaattg acggctcgag ggagtagcat agggtttgca gaatccctgc ttcgtccatt
     4201 tgacaggcac attatgcatc gatgataagc tgtcaaacat gagcagatcc tctacgccgg
     4261 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     4321 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcaaat attttatctg
     4381 aggtgcttcc tcgctc
//
