LOCUS       sequence-423567-        4387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Saccharothrix texasensis LRD-modifying cytochrome P450
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4387)
  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 4387)
  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: pCYP41
FEATURES             Location/Qualifiers
     source          1..4387
                     /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(4299..4318)
                     /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 tgtttatctc tataccctat
     2101 taaggaggtc taatatgttg actaccgtgg ctgtggcacc ggaaccccgt cagtacccgt
     2161 ttgaaccatt caccggcgaa ctgagcccgg agctggttgg catggtcgaa accgatccgg
     2221 tgtctcgcgt gttactgccg gacggtgaac cggcgtggct ggtactcgat tatgctggct
     2281 gcgtgaaagt gctggctgat ccacgtttct cgcgcgttct tcatcggcca cctcccggtc
     2341 atccgcctct gcgcgatctg aatcaggacg gcgccgcgca cgccgctgtg cggagagttg
     2401 gtagtcgtgc cttcggtggt cgccgcatgg cgcattaccg cccacgcgtg caagcactgg
     2461 tcgatgaatt ggtcgacgcc atgatcgccg ggccccgtcc agcggatctt gtaagtggac
     2521 tggtggcccc tctgccattg ctggtgattt gcgaagtgat tggcgttcca gcaacagacc
     2581 gcgaacgatt tcatggctgg ctggcgggac tgaatagtgt ggtagactac aactcggcgg
     2641 aggcggcgaa aagcctgtgg gtgtatctgg cggggcaatt agcagcgaag cgtgttgaac
     2701 cgggcgatga cttattgtca gtgtggttgg aaggtggtga cgatcacggg ctttctgatg
     2761 aggagatcat cctgctggcc atgggtttac tgcgcggggg tctggaagta gggagcacag
     2821 ctgccggagt ccgtgtgttg tttcagcatc ctgaacagtt agctgatctg gtgcgctcgc
     2881 cagagaaaat tccggctgcg gttgatgaaa ttctgcgtta cactgctgtg agttccatgt
     2941 tccgagtcca agttgtcacg gaagatgtta ctgtcggcga tgttgcaatg agcgccggtg
     3001 atcgggttat ggcagtgcca tgggtcgcaa atcgtgatcc acgcgtgttc tccgacccga
     3061 atgtatttga cattgaccgc gtaccgcgta ctgcgcatct ggcgtttggt ttcggcccgc
     3121 acttctgcct gggtactgcc ctggctcgga tggaggtgga actttccata gcgaccctgc
     3181 tgcgtcgcct gccaggtctg gcccctgctg ttgcggttga agaattacct tggcgccacg
     3241 atcggattaa tggcgggatc gttgagttcc cggtcgtctg gagagaggat cagtaataag
     3301 gatgagctct acaaataagc agaggtggtt gtgttgcgaa aaaaaaaaaa acaccctaac
     3361 gggtgttttt tttttttggt gtccccacgt gtggcgctgg agaccgtcca atggcggcgc
     3421 gccatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     3481 ttcagggtgg tgaatatgag cccgaaacgt cgtacccagg cagaacgtgc aatggaaacc
     3541 cagggtaaac tgattgcagc agcactgggt gttctgcgtg aaaaaggtta tgcaggtttt
     3601 cgtattgcag atgttccggg tgcagccggt gttagccgtg gtgcacagag ccatcatttt
     3661 ccgaccaaac tggaactgct gctggcaacc tttgaatggc tgtatgagca gattaccgaa
     3721 cgtagccgtg cacgtctggc aaaactgaaa ccggaagatg atgttattca gcagatgctg
     3781 gatgatgcag cagatttttt tctggatgat gattttagca tcggcctgga tctgattgtt
     3841 gcagcagatc gtgatccggc actgcgtgaa ggtattctgc gtaccgttga acgtaatcgt
     3901 tttgttgttg aagatatgtg gctgggtgtg ctggtgagcc gtggtctgag ccgtgatgat
     3961 gccgaagata ttctgtggct gatttttaac agcgttcgtg gtctgacagt tcgtagcctg
     4021 tggcagaaag ataaagaacg ttttgaacgt gtgcgtaata gcaccctgga aattgcacgt
     4081 gaacgttatg caaaattcaa acgttgataa ggatcctaat tggtaacgaa tcagacaatt
     4141 gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacaggca
     4201 cattatgcat cgatgataag ctgtcaaaca tgagcagatc ctctacgccg gacgcatcgt
     4261 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     4321 tggggaagat cgggctcgcc acttcgggct catgagcaaa tattttatct gaggtgcttc
     4381 ctcgctc
//
