LOCUS       sequence-164479-        5405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P450 BM3 variant (A74G/C773S/C810S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5405)
  AUTHORS   Tan CY, Hirakawa H, Suzuki R, Haga T, Iwata F, Nagamune T
  TITLE     Immobilization of a Bacterial Cytochrome P450 Monooxygenase System 
            on a Solid Support.
  JOURNAL   Angew Chem Int Ed Engl. 2016 Oct 26. doi: 10.1002/anie.201608033.
  PUBMED    27781345
REFERENCE   2  (bases 1 to 5405)
  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: "Angew Chem 
            Int Ed Engl. 2016 Oct 26. doi: 10.1002/anie.201608033."
COMMENT     Sequence Label: pBsaBM3
FEATURES             Location/Qualifiers
     source          1..5405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..192
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             193..852
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(259..278)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    1060..1081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1096..1101,1102..1119,1120..1126)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1134..1150
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1139..1161
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1158..1174
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1158..1174
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4422..4439)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4422..4438)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4431..4453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4593..4610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4764..5352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4844..4863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggaagatc acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc
       61 tgggccaact tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac
      121 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
      181 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
      241 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
      301 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
      361 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
      421 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
      481 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
      541 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
      601 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
      661 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
      721 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt
      781 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
      841 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg tgctacgcct
      901 gaataagtga taataagcgg atgaatggca gaaattcagc ttggcccagt gccaagctcc
      961 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1021 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1081 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1141 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgaattcg agctcgggtc
     1201 tcgggaggta gcatgacaat taaagaaatg cctcagccaa aaacgtttgg agagcttaaa
     1261 aatttaccgt tattaaacac agataaaccg gttcaagctt tgatgaaaat tgcggatgaa
     1321 ttaggagaaa tctttaaatt cgaggcgcct ggtcgtgtaa cgcgctactt atcaagtcag
     1381 cgtctaatta aagaagcatg cgatgaatca cgctttgata aaaacttaag tcaaggcctt
     1441 aaatttgtac gtgattttgc aggagacggg ttatttacaa gctggacgca tgaaaaaaat
     1501 tggaaaaaag cgcataatat cttacttcca agcttcagtc agcaggcaat gaaaggctat
     1561 catgcgatga tggtcgatat cgccgtgcag cttgttcaaa agtgggagcg tctaaatgca
     1621 gatgagcata ttgaagtacc ggaagacatg acacgtttaa cgcttgatac aattggtctt
     1681 tgcggcttta actatcgctt taacagcttt taccgagatc agcctcatcc atttattaca
     1741 agtatggtcc gtgcactgga tgaagcaatg aacaagctgc agcgagcaaa tccagacgac
     1801 ccagcttatg atgaaaacaa gcgccagttt caagaagata tcaaggtgat gaacgaccta
     1861 gtagataaaa ttattgcaga tcgcaaagca agcggtgaac aaagcgatga tttattaacg
     1921 catatgctaa acggaaaaga tccagaaacg ggtgagccgc ttgatgacga gaacattcgc
     1981 tatcaaatta ttacattctt aattgcggga cacgaaacaa caagtggtct tttatcattt
     2041 gcgctgtatt tcttagtgaa aaatccacat gtattacaaa aagcagcaga agaagcagca
     2101 cgagttctag tagatcctgt tccaagctac aaacaagtca aacagcttaa atatgtcggc
     2161 atggtcttaa acgaagcgct gcgcttatgg ccaactgctc ctgcgttttc cctatatgca
     2221 aaagaagata cggtgcttgg aggagaatat cctttagaaa aaggcgacga actaatggtt
     2281 ctgattcctc agcttcaccg tgataaaaca atttggggag acgatgtgga agagttccgt
     2341 ccagagcgtt ttgaaaatcc aagtgcgatt ccgcagcatg cgtttaaacc gtttggaaac
     2401 ggtcagcgtg cgtgtatcgg tcagcagttc gctcttcatg aagcaacgct ggtacttggt
     2461 atgatgctaa aacactttga ctttgaagat catacaaact acgagctgga tattaaagaa
     2521 actttaacgt taaaacctga aggctttgtg gtaaaagcaa aatcgaaaaa aattccgctt
     2581 ggcggtattc cttcacctag cactgaacag tctgctaaaa aagtacgcaa aaaggcagaa
     2641 aacgctcata atacgccgct gcttgtgcta tacggttcaa atatgggaac agctgaagga
     2701 acggcgcgtg atttagcaga tattgcaatg agcaaaggat ttgcaccgca ggtcgcaacg
     2761 cttgattcac acgccggaaa tcttccgcgc gaaggagctg tattaattgt aacggcgtct
     2821 tataacggtc atccgcctga taacgcaaag caatttgtcg actggttaga ccaagcgtct
     2881 gctgatgaag taaaaggcgt tcgctactcc gtatttggat gcggcgataa aaactgggct
     2941 actacgtatc aaaaagtgcc tgcttttatc gatgaaacgc ttgccgctaa aggggcagaa
     3001 aacatcgctg accgcggtga agcagatgca agcgacgact ttgaaggcac atatgaagaa
     3061 tggcgtgaac atatgtggag tgacgtagca gcctacttta acctcgacat tgaaaacagt
     3121 gaagataata aatctactct ttcacttcaa tttgtcgaca gcgccgcgga tatgccgctt
     3181 gcgaaaatgc acggtgcgtt ttcaacgaac gtcgtagcaa gcaaagaact tcaacagcca
     3241 ggcagtgcac gaagcacgcg acatcttgaa attgaacttc caaaagaagc ttcttatcaa
     3301 gaaggagatc atttaggtgt tattcctcgc aactatgaag gaatagtaaa ccgtgtaaca
     3361 gcaaggttcg gcctagatgc atcacagcaa atccgtctgg aagcagaaga agaaaaatta
     3421 gctcatttgc cactcgctaa aacagtatcc gtagaagagc ttctgcaata cgtggagctt
     3481 caagatcctg ttacgcgcac gcagcttcgc gcaatggctg ctaaaacggt cagcccgccg
     3541 cataaagtag agcttgaagc cttgcttgaa aagcaagcct acaaagaaca agtgctggca
     3601 aaacgtttaa caatgcttga actgcttgaa aaatacccgg cgagcgaaat gaaattcagc
     3661 gaatttatcg cccttctgcc aagcatacgc ccgcgctatt actcgatttc ttcatcacct
     3721 cgtgtcgatg aaaaacaagc aagcatcacg gtcagcgttg tctcaggaga agcgtggagc
     3781 ggatatggag aatataaagg aattgcgtcg aactatcttg ccgagctgca agaaggagat
     3841 acgattacgt gctttatttc cacaccgcag tcagaattta cgctgccaaa agaccctgaa
     3901 acgccgctta tcatggtcgg accgggaaca ggcgtcgcgc cgtttagagg ctttgtgcag
     3961 gcgcgcaaac agctaaaaga acaaggacag tcacttggag aagcacattt atacttcggc
     4021 tgccgttcac ctcatgaaga ctatctgtat caagaagagc ttgaaaacgc ccaaagcgaa
     4081 ggcatcatta cgcttcatac cgctttttct cgcatgccaa atcagccgaa aacatacgtt
     4141 cagcacgtaa tggaacaaga cggcaagaaa ttgattgaac ttcttgatca aggagcgcac
     4201 ttctatattt gcggagacgg aagccaaatg gcacctgccg ttgaagcaac gcttatgaaa
     4261 agctatgctg acgttcacca agtgagtgaa gcagacgctc gcttatggct gcagcagcta
     4321 gaagaaaaag gccgatacgc aaaagacgtg tgggctgggt aaggatctga gaccgtaccc
     4381 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cactggccgt cgttttacaa
     4441 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4501 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4561 agcctgaatg gcgaatgagc ttcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4621 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4681 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4921 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5161 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaactcc
     5401 gtcga
//
