LOCUS       sequence-423277-        4655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Scytonema sp. HK-05 LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4655)
  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 4655)
  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: pCYP22
FEATURES             Location/Qualifiers
     source          1..4655
                     /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(4567..4586)
                     /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 tgtttaggac ctagcagaag
     2101 gagaataaaa tggaaatcaa ggaaggctat tttaagcagc aacaaaccct caaaaatgcc
     2161 gataccgccc aggaacacca aattaaccgc ccatcgccgt tcgatccgtc ccccaccccg
     2221 aaaacctgtc cagtcgtgcg tggttaccca tttgtaggcg tactgccaga actgctgcag
     2281 aacccgctgc gcctgtttca gcgggcagtg aaagaccatc cgggagaaat tgttgccctg
     2341 aaactggggc caattcaagc ccatttagta acacacccgg atcacgtgaa atatgtgttg
     2401 aatgataatt ggcagaattt tgggaaaggt gaagtcatgt ggaaaccagc ccgtcaggtc
     2461 gtcggtaacg gcctgatcac aagccacggc gaaagttggt ttcgcaatcg cagactgatg
     2521 cagccgatgt ttgcggcgaa tcaggctaac gcgctgaccg agctgtcagt taaacttatc
     2581 gctgaacagc tcgcacgttt tgaaaccaca gcaaattcag aacctatcga cattaagaac
     2641 gaaatggcac tgttaacgca gacggtgatc ctgggcgtta tattctcctc tagcattgat
     2701 cgttccaaag cagaaacgct caccgaagct ctgaaagttc tggttcgaga aaccagtcgt
     2761 agaatgtttc tgtgttttct gccaaaacag tttccgatcc cgggtgaacg cgcgttccgc
     2821 aatgcgattc aaactacaga tgatacaatt ctgtcaatca ttcgcgcccg gcgttggtct
     2881 agccttaact acggagactt gtttgagaga ttactgttta gccgtgatga ggaatcaaaa
     2941 accggaatga acgaccagca actgcgggac gagagcatta cactgttcct cgcaggatat
     3001 gaaaccacga gcaccgcact gacgtgggtt tggtatttgt tggacaaaca ccccgaagta
     3061 cagagtaaac tccgcgccga aattgacgct gttattggga aacgccagcc cacttgtgct
     3121 gatgtagcga aactgcagta caccaaaatg gttattcaag aagtgatgcg actctacccg
     3181 cccgtgctgt ttctggcgcg ggtagtggaa gctgacgacg tgattgatgg ctacccgctg
     3241 aaagctggca caaccgtcgt cctgtgtaag tacctgactc atcgtctgcc tgaattttgg
     3301 gatcaaccgg aagtctttga tccagagcgt ttcgcatcag aatatagtac acagcgtcat
     3361 tcgtattcct attttccttt cggcggtggt ccacgttatt gcattgggaa acatttggca
     3421 cttctcgaga tgcagcttat tgtcgctatg atggtacaaa agtatcgttt gcgttgcgtc
     3481 cccggccatc cggtggaggc gagtcctaaa attatcctgc accctcgcta cggcctcccg
     3541 atgactgtgg aaccggttta ataaggatga gctctacaaa taagcagagg tggttgtgtt
     3601 gcgaaaaaaa aaaaaaaaca ccctaacggg tgtttttttt tttttggtgt ccccacgtgt
     3661 ggcgctggag accgtccaat ggcggcgcgc catcgaatgg tgcaaaacct ttcgcggtat
     3721 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgagcc cgaaacgtcg
     3781 tacccaggca gaacgtgcaa tggaaaccca gggtaaactg attgcagcag cactgggtgt
     3841 tctgcgtgaa aaaggttatg caggttttcg tattgcagat gttccgggtg cagccggtgt
     3901 tagccgtggt gcacagagcc atcattttcc gaccaaactg gaactgctgc tggcaacctt
     3961 tgaatggctg tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa aactgaaacc
     4021 ggaagatgat gttattcagc agatgctgga tgatgcagca gatttttttc tggatgatga
     4081 ttttagcatc ggcctggatc tgattgttgc agcagatcgt gatccggcac tgcgtgaagg
     4141 tattctgcgt accgttgaac gtaatcgttt tgttgttgaa gatatgtggc tgggtgtgct
     4201 ggtgagccgt ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga tttttaacag
     4261 cgttcgtggt ctgacagttc gtagcctgtg gcagaaagat aaagaacgtt ttgaacgtgt
     4321 gcgtaatagc accctggaaa ttgcacgtga acgttatgca aaattcaaac gttgataagg
     4381 atcctaattg gtaacgaatc agacaattga cggctcgagg gagtagcata gggtttgcag
     4441 aatccctgct tcgtccattt gacaggcaca ttatgcatcg atgataagct gtcaaacatg
     4501 agcagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     4561 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     4621 tgagcaaata ttttatctga ggtgcttcct cgctc
//
