LOCUS       sequence-423278-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Micromonospora humi LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4577)
  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 4577)
  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: pCYP28
FEATURES             Location/Qualifiers
     source          1..4577
                     /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             2100..2108
                     /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(4489..4508)
                     /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 tgtttaggat cttagtaact
     2101 aaggaggtaa ttatttatga gcatcacaac ccatgctcca agacgtgaag ctcgcccacc
     2161 gcgagtacct gctgggctgc cgcttgtggg tcatggtctg cagttattac gtcgccggct
     2221 ggattttctg caggaagcac gtcgctatgg tccagtggtg agagttggac tggggccttc
     2281 aacggcctat ctggtaaatg accataccct tctccaccag atgctggtta atgatgccgc
     2341 actgtttccg cgcggcatcc atttccgtaa agcgcgcgcg gtagctggtg atgggatcat
     2401 tactgcagaa ggccccgcgc atcgtcgtca acgtcgtctt gtgctgcctg ctttttcacg
     2461 taaacgtgtg catggttatg gcgatgtgat gcgtcgtatt gcgatggaac gaatctcatc
     2521 ctggccagag gacagaccta ttgaattgaa accggaattc gtatcgctgg caacgaccat
     2581 cggtacccgc gccctcttct caaccgatct gggcgcttcg gctgtctcgg tcattgaacg
     2641 cgcgctgcct gtactgacga gaggcgtaag tttgcgggct atggacccaa ccgggttagt
     2701 tgagcactta ccgctgccgg caaatagacg atttggagct acaatgcgcg aattacacgc
     2761 agtgattgac ggcatcatcg cttcgtatcg tcgcaccgag cagcagggtg atgatctgct
     2821 gtccctgctg atgttagcta ctgatgaagg ggatggcaaa ggtatgaccg atcggcagct
     2881 ccgtgacgaa gttatatcaa ttctcctttc ttctgcagaa acgaccgcgc tgacgatgag
     2941 ctgggcatgt catgagctgg gacgtcaccc agaggtattt cgctcactgc gggcggaggt
     3001 ggatcgagtc ctcccagacg gtgaacctcc ccgtcatgag cagctgcccg agttagaatt
     3061 aacccgtcgc gtagtgaagg aaacgctgcg cctctacccg ccaacgtatt tcctgtcccg
     3121 tacggccacg aaagacgtcc gtctgggcca gcatttgatc ccgggtggct caaccgttct
     3181 gtactcattc tactcccaac atcgcgatcc tcgcttgttc agagatccgc atacgtttga
     3241 tcctgaccgg tggttgccag agcgtgcggc tgacgtcacg cctgaagcgt acatgccgtt
     3301 tggtgaaggc gcccaccggt gcgtgggtga aggctttgca tgggcggaag ctatgacgag
     3361 cctggcgatt atagcgttgc accgtgatct tagaccggtt gacggtgatc aagtgcgtcc
     3421 cacaggtacc gtgaccttga tgcctcgggg tctgtcaatg gtcctgcccc gtagagcagc
     3481 gtaataagga tgagctctac aaataagcag aggtggttgt gttgcgaaaa aaaaaaaaaa
     3541 caccctaacg ggtgtttttt tttttttggt gtccccacgt gtggcgctgg agaccgtcca
     3601 atggcggcgc gccatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     3661 agagagtcaa ttcagggtgg tgaatatgag cccgaaacgt cgtacccagg cagaacgtgc
     3721 aatggaaacc cagggtaaac tgattgcagc agcactgggt gttctgcgtg aaaaaggtta
     3781 tgcaggtttt cgtattgcag atgttccggg tgcagccggt gttagccgtg gtgcacagag
     3841 ccatcatttt ccgaccaaac tggaactgct gctggcaacc tttgaatggc tgtatgagca
     3901 gattaccgaa cgtagccgtg cacgtctggc aaaactgaaa ccggaagatg atgttattca
     3961 gcagatgctg gatgatgcag cagatttttt tctggatgat gattttagca tcggcctgga
     4021 tctgattgtt gcagcagatc gtgatccggc actgcgtgaa ggtattctgc gtaccgttga
     4081 acgtaatcgt tttgttgttg aagatatgtg gctgggtgtg ctggtgagcc gtggtctgag
     4141 ccgtgatgat gccgaagata ttctgtggct gatttttaac agcgttcgtg gtctgacagt
     4201 tcgtagcctg tggcagaaag ataaagaacg ttttgaacgt gtgcgtaata gcaccctgga
     4261 aattgcacgt gaacgttatg caaaattcaa acgttgataa ggatcctaat tggtaacgaa
     4321 tcagacaatt gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat
     4381 ttgacaggca cattatgcat cgatgataag ctgtcaaaca tgagcagatc ctctacgccg
     4441 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     4501 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcaaa tattttatct
     4561 gaggtgcttc ctcgctc
//
