LOCUS       sequence-413666-        6862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of biformene in E. coli MEV20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  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 6862)
  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: pLRD09
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(90..133)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    136..157
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        237..255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    256..280
                     /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)."
     promoter        2117..2135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2136..2160
                     /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)."
     misc_feature    3410..3430
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3431..3488
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3512..3529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4063..4285
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4333..5283
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFWIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(5897..6688,6689..6757))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6520..6539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6758..6862)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
      121 taaggatgat ttctggaatt cgcggccgct tctagagcgc gaagacttaa tcaaatccgt
      181 agcagtgcac aagcaagtga gtgctgacga gctcaggagc cgcaatacac aggtgctaat
      241 acgactcact ataggggaat tgtgagcgga taacaattcc cctctaaacg ccagccacta
      301 aagcggtcaa cgcatgtgct ttgcgttctg atgagacagt gatgtcgaaa ccgcctctac
      361 aaataatttt gtttaagaag gtcaaacaca ttagcaacga taagggggtc tttatgtcgt
      421 ccatcagcca tcaagcggct ccgacgcgtg ctcagcacac caattcctac gtgcagctgg
      481 cgcgccaatt agtgacttca gtcgacaatg acccatgggg tgacgtcccg cctagtgtgt
      541 acgaaaccgc acgggtcaca tcgtgggctc cgtggctgga aggtcacgag cgccgcctgg
      601 catggttact cgaacgccaa agtgcggccg gttcctgggg tgaaggtccg accccttatc
      661 gtctgcttcc tacactgtct gttactgaag cacttctcag caccctgcgt cagaacacag
      721 ccgccggtgt ttctcgcgaa cgcctggccg cggcagtcga caatggtctt gcagcgttgc
      781 gtgatctgag cggtaccggc ggctggccgg ataccgccgc catcgagatt ctggcacctg
      841 atctggtggt tttaatcaat gaccatctgg atcaaccaga agttgccgca cttccgcgcc
      901 tgggtccttg ggcccgcggt caaagattag cccaacctca cggttttcaa gcggcgctcc
      961 ctgaccgcgt tgctgaacgt tgccaggtgg ctgggggcgt tccgctgaaa cttcatcaca
     1021 cttttgaagg cgtggcacgc cgcctgccgc gtatggtgcc tggagttccc ggtggactcc
     1081 tgggctcaag tccagcggct acggcagcgt ggttagcaac cggtccggac gaagggcgtg
     1141 accaagcagt tacggctctt acggcggttg cggaacgcta cgatggcctg ttcccagaag
     1201 caaccccgat tagtgtgttc gaacgcctgt ggatctctgt agcattagct cgcccgggtc
     1261 tgcccgccgc gtgcgtacct acaattcgtg cctgggccgc tgaaatctat gatgcgaccg
     1321 gagtgcgtgg tgctcccggt ttactgccag atactgatga tacagcgatg gcggtcttag
     1381 cgtctgcact ggccggtagc ccgcgagatc cttctccact gtcggcattt gaggcgggtg
     1441 atcactatga ttgttacgtg ggcgaggaca ccggtagctc tacggctaat gcgcacgcac
     1501 tccaggctct tacggcgtgg cttagccacc gtccggccac tggcgacgct ttacaggcca
     1561 gacgcgatct gacgcgagat tggctgctgg cccagcagga aagtgatggg gcgtggcgtg
     1621 ataaatggca tgcgagccca tattatgcaa ctgaacgttg cgttactgcc ctgtcaggcc
     1681 atacgggccc tacgacgcgt gatgctatcc gctcggctgc cgattgggtc ctggatgcac
     1741 agtccgatga tggcagttgg ggtgtatggg gaggtaccgc ggaagaaacc gcctacgccg
     1801 tgaacatctt gcttaacagc ccggaccata ccgggacgcc agaagcgaca caggcgttaa
     1861 aactggcaga aaatgtattg cgtgaagcgg tgcattcgtc cggtcaccat catcccgctt
     1921 tatggcatga caaaaccctt tacgcgcccc aagcgatggc acaggcggaa gtcattgcag
     1981 ccttggaact gctgcaggca cgtcgtccgt aataattatg ccttccgcta atcaactcgg
     2041 taccaaattc cagaaaagag acgctttcga gcgtcttttt tcgttttggt ccgtggacaa
     2101 gcacagcata gccaagtaat acgactcact ataggggaat tgtgagcgga taacaattcc
     2161 cctctaggtc cgaagatact tagggtgtct caaggtgcgt accttgactg atgagtccga
     2221 aaggacgaaa cacccctcta caaataattt tgtttaagag agagcctgaa ttaccaataa
     2281 ggaggaatta atgacccgca cgggcgatgc tgtgacgatt ttgccacagc cagattttac
     2341 cgccaccttc cctggtccct tcccgaccag cccccacggt gagcgtacag agcgtcagtt
     2401 gttaggctgg ctggaagagt atccgctgct cccttcggcg cgcgctcgat cagttctggt
     2461 aaatattaca agccatggtg ttagtcgcac ccttccgacc gccgatgctg atgatctcgt
     2521 cctgtttgcg gaattacttc tctggctgac agccttcgac gatatgcatg gtgagtcaaa
     2581 cgccgcacgc gatctggtcg ctcttgtgga ccgcaccgcc gagttaacac tggttttagc
     2641 aggcggtagt ccgcctcccc tgaccaaccc gtttccggcg gctctctacg atctgttggc
     2701 gcgttttcga gcacgtaccg gtcctgcggc ttatcttcgc ttggccgcgt cgctgcgtga
     2761 cacaattatg gcgttagtgt gggaagctca tcacgttgca gaaccggagc gcgtagcact
     2821 ggaaacgtac ctggagatgc gtccgcatac cgtgtttgtc cggacgattt tcgccgcggc
     2881 cgaaatcgtg ttggactacg aactgacaga tgcacagcgg gcgctggcgc cggtcagaca
     2941 tctggaaacc gcagtagcca atctggctgg atggattaac gatctcgcgt cttatgaacg
     3001 ggaagcagcg cgcggaccgg cgcaaccgtt gtcactccca accttgctgc gcgctcgaca
     3061 cggtggtagc ttagaagaag cattcgcgcg ggccggcggt atgtgcgaaa atgaggccgc
     3121 agtcgccaga caaggcatta ccagtcttgc aggggaccca ccgagcgctt taaccgcgca
     3181 cgcgcgcgct ctggaagata ttgcgcgcag ctttgtgtgg catacgagcc acgcccgata
     3241 ccagggtccc aaacgcggcg ctgcaccaac gagtcggtaa taagcctcgt tgctcttcat
     3301 aaatcatccc tatgacccct tcaaaaaaaa aaaacaccct aacgggtgtt tttttttttt
     3361 tggtctgcct ctcgctgacg gtcacaatgc gcttacgaaa gtcttcctgt actagtagcg
     3421 gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac
     3481 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
     3541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatgacagta agacgggtaa
     3601 gcctgttgat gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacg
     3661 ggataatccg aagtggtcag actggaaaat cagagggcag gaactgctga acagcaaaaa
     3721 gtcagatagc accacatagc agacccgcca taaaacgccc tgagaagccc gtgacgggct
     3781 tttcttgtat tatgggtagt ttccttgcat gaatccataa aaggcgcctg tagtgccatt
     3841 tacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga aaaagacagc
     3901 gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg
     3961 agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc aaacagcgtt
     4021 tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata cacagggctg
     4081 ggatctattc tttttatctt tttttattct ttctttattc tataaattat aaccacttga
     4141 atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct agcagaattt
     4201 acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca aaggaaaagg
     4261 actagtaatt atcattgact agcccatctc aattggtata gtgattaaaa tcacctagac
     4321 caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc gattagtcgc
     4381 tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact actcaacccc
     4441 acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa ccaatacgct
     4501 cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc aaccagagag
     4561 ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggcttttg gattttccag
     4621 tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag tgaagagata
     4681 ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca tgttaagtct
     4741 tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact aacacaaaag
     4801 aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat gttaatgctt
     4861 gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc aataagtaaa
     4921 gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg cccgactgat
     4981 acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga acttgagaac
     5041 aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga ttcctaccta
     5101 cataacggac taagaaaaac actacacgat gctttaactg caaaaattca gctcaccagt
     5161 tttgaggcaa aatttttgag tgacatgcaa agtaagtatg atctcaatgg ttcgttctca
     5221 tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata cggaaggatc
     5281 tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa acaaaagtag
     5341 aacaactgtt caccgttaca tatcaaaggg aaaactgtcc atatgcacag atgaaaacgg
     5401 tgtaaaaaag atagatacat cagagctttt acgagttttt ggtgcattca aagctgttca
     5461 ccatgaacag atcgacaatg taacagatga acagcatgta acacctaata gaacaggtga
     5521 aaccagtaaa acaaagcaac tagaacatga aattgaacac ctgagacaac ttgttacagc
     5581 tcaacagtca cacatagaca gcctgaaaca ggcgatgctg cttatcgaat caaagctgcc
     5641 gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa tcatggcaat tctggaagaa
     5701 atagcgcttt cagccggcaa accggctgaa gccggatctg cgattctgat aacaaactag
     5761 caacaccaga acagcccgtt tgcgggcagc aaaacccgta ccgattatca aaaaggatct
     5821 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5881 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5941 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6001 gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag
     6061 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6121 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6181 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6241 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6301 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6361 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6421 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6481 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6541 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6601 taccgctgtt gagatccagt tcgatataac ccactcgtgc acccaactga tcttcagcat
     6661 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6721 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6781 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6841 ataaacaaat aggggttccg cg
//
