LOCUS       sequence-413664-        8306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of (16R)-ent-kauran-16-ol in E. coli MEV20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8306)
  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 8306)
  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: pLRD07
FEATURES             Location/Qualifiers
     source          1..8306
                     /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        178..196
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    197..221
                     /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        1752..1770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1771..1795
                     /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        3630..3648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3649..3673
                     /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)."
     RBS             3781..3789
                     /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)"
     misc_feature    4854..4874
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4875..4932
                     /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(4956..4973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5507..5729
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5777..6727
                     /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(7341..8132,8133..8201))
                     /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     7964..7983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8202..8306)
                     /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 tcaaatctaa
      181 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctacgt tttcaaggtg
      241 gtagactgtc gccggatgtg tatccgacct gacgatggcc caaaagggcc gaaacagtcc
      301 tctacaaata attttgttta agagaatacc ccgctaagaa ggtaattatg tccatgcaaa
      361 gtttagataa gaattttcga catttatctc gtaaagaaaa gttacaacaa ttggttgata
      421 agcaatggtt atcagaagaa caattcgaca ttttactgaa tcatccatta atcgatgaag
      481 aagtagccaa tagtttaatt gaaaatgtca tcgcgcaagg tgcattaccc gttggattat
      541 taccgaatat cattgtggac gataaggcat atgttgtacc tatgatggtg gaagagcctt
      601 cagttgtcgc tgcagctagt tatggtgcaa agctagtgaa tcagactggc ggatttaaaa
      661 cggtatcttc tgaacgtatt atgataggtc aaatcgtctt tgatggcgtt gacgatactg
      721 aaaaattatc agcagacatt aaagctttag aaaagcaaat tcataaaatt gcggatgagg
      781 catatccttc tattaaagcg cgtggtggtg gttaccaacg tatagcgatt gatacatttc
      841 ctgagcaaca gttactatct ttaaaagtat ttgttgatac gaaagatgct atgggcgcta
      901 atatgcttaa tacgatttta gaggccataa ctgcattttt aaaaaatgaa tttccgcaaa
      961 gcgacatttt aatgagtatt ttatccaatc atgcaacagc gtccgttgtt aaagttcaag
     1021 gcgaaattga tgttaaagat ttagcaaggg gcgagagaac tggagaagag gttgccaaac
     1081 gaatggaacg tgcttctgta ttggcccaag tagatattca tcgtgcagca acacataata
     1141 aaggtgttat gaatggcata catgctgttg ttttagcaac aggaaatgat acgcgtggtg
     1201 cagaagcaag tgcgcatgca tacgcgagtc gtgacggaca gtatcgtggt attgctacat
     1261 ggcgttacga tcaagatcgt caacgattga ttggtacaat tgaagtgcct atgacattgg
     1321 caatcgttgg cggtggtaca aaagtattac caattgctaa agcttcatta gagctactaa
     1381 atgtagagtc agcacaagaa ttaggtcatg tagttgctgc cgttggttta gcgcaaaact
     1441 ttgcagcatg tcgcgcgctt gtgtcagaag gtattcaaca aggtcatatg agtttacaat
     1501 ataaatcatt agctatcgtt gtaggggcaa aaggtgatga aattgctaaa gtagctgaag
     1561 ctttgaaaaa agaaccccgt gcaaatacac aagcagcgga acatatttta caagaaatta
     1621 gacaacaata ataacggatg taaacagaga aacgtctaac taaaaaggcc tcccaaatcg
     1681 gggggccttt tttattgata acaaaagcct aatacacctg ttcgtgacga ctacggatgc
     1741 gggcaaggtg ctaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     1801 aaacgccagc cactaaagcg gtcaacgcat gtgctttgcg ttctgatgag acagtgatgt
     1861 cgaaaccgcc tctacaaata attttgttta agagattatt gattaatttt aaggaggatt
     1921 tatatgctgg aagttccggc acagccaacg ccagcgcctc gtgaagcaga agcggcagcc
     1981 ttattggcag cgaccgttgc tgatccttgg ggcttagtcg cacctagtgt gtacgacaca
     2041 gctcgtctgg taagccttgc cccgtggctg gatggacacc gtgaacgcct gggttacctc
     2101 gtgaaagaac agaatcagga tggctcatgg ggcgctccgg acggctatgg tctggtccct
     2161 accttgtcgg ccgttgaagc cctgctgacg gaattagcgc gcaccgatag tggagcgccg
     2221 catcctccac acgatgatct ggctgcggcc tgcgcgggcg gacttggtgc gttacaggat
     2281 ggcctcttgg ctgggccggt tcctgatact atcggcgttg agtttgttgc gccttcatta
     2341 ctggcggata tcaacacgcg tctggccgca ctgacagaac aagcccctgg caaactgggc
     2401 gcatggtccg gtacaaccct gacgtcacca gctccggatc tggatggcgc acttctggcc
     2461 ggcgttcgtg agatgactga gcaggcgcca ctgccagaga aactgtggca taccttggaa
     2521 gccatcaccc gcgatggtac ccgtggggca cgcccccatg aaggtgcacc gccacacaat
     2581 gggtctgttg gttgtagtcc agcagctacc gctgcatggc ttggtgcgtc gcctgatcct
     2641 gcagcgccag gtgtagcata cctgcgtgat gttcaggctc gctttggcgg cccggtgccg
     2701 tccattacgc cgatcgtgta ctttgagcag gcatgggtgc tgaatagttt agccgcatcg
     2761 ggactgcgct atgaagcgcc tgcagcatta ctggacagtc tggaagccgg cttgactgat
     2821 gaaggcatcg cggcagcacc gggcctccct tctgacagcg acgataccgc cgcagtatta
     2881 ttcgcgctgg cccagcacgg tcgtacacat cgcccagact ccctgatgca cttccgccgt
     2941 gatggctatt tttcatgttt tggtgtcgag cgcacgccat caacctcaac aaacgcgcat
     3001 atactggaag cgttgggaca ccatgttaca gtgcgccccg acgatgctgg gcgttatggc
     3061 gccgaaatcc gtatgatttc tgactggctg ctggataatc aactgccaga tggctcttgg
     3121 atggataaat ggcacgcgtc accctattac gccaccgcct gctgtgccct tgctctggcc
     3181 gaatttggcg gaccgagcgc tcgtgctgca gtggatcgtg cggcagcgtg ggctctggaa
     3241 actcaacgtg ccgatggcag ctggggtcgc tggcagggca caacagaaga aaccgcatac
     3301 atggtgcagc tcttgatgcg cacccgcacg cctggttcgc caggtactgt ggctagatcg
     3361 gctgcgcgcg gctgtgatgc cctcctggct catgatgatc cggcctcgta cccgggcctt
     3421 tggcatgata aagacattta tgccccggta acggtgattc gagcggcacg tttagccgcg
     3481 ctggctctgg gaggtgcaga gtctgccgcg agtggtgggg cgtaataatt atgccttccg
     3541 ctaatcaact cggtaccaaa ttccagaaaa gagacgcttt cgagcgtctt ttttcgtttt
     3601 ggtccgtgga caagcacagc atagccaagt aatacgactc actatagggg aattgtgagc
     3661 ggataacaat tcccctctag gtccgaagat acttagggtg tctcaaggtg cgtaccttga
     3721 ctgatgagtc cgaaaggacg aaacacccct ctacaaataa ttttgtttaa gactggagta
     3781 taaggaggtt tattatgaac aacaagctgg tgagtcatct gcttgatagt gccgatcccg
     3841 ctgtacgggc aaaagctatt gaagccgccg ccctggcccg ccagtgggca cgtccagtac
     3901 aacaagagtt gtcggataag gtctacgata ccctggcggc gacggtgtgt gtgattgcgc
     3961 ccgagctgga cgtcgccgat catgctttac ttgtggagta ctcgctttgg ctgtacttat
     4021 tggatggtcg gctggatgat ttcgaacatt atgggacccg gccggaggat gttggccgac
     4081 gtgttatcgc cgtgttgcgc ggcggtcggg cagaagcccg tgcagacgac ttctttgaaa
     4141 cgtcgctggc cgctttggtg gaagaattac gaacgcgcga tggctgctgt ggactgcttg
     4201 aacgattcgt gctgcgcctg gtagacgggg ttcgcgcggg tgtacgtcaa gcggtgctca
     4261 gtcgccgtat cgcggaaggt gccgaaccgc tgccgacgat ggaagatttt ctggagctgg
     4321 cttaccgtca tgtcaattat cgtagcgtcg ccctggcgtt gctgatcacc gtgggagagc
     4381 gtccagatag tgctgctcag gaaagactgg atgctgctct ggtgcctgct tcgcgcgcgg
     4441 tacgccttgc gaacgatctg cgcacctggg ccaaagacgc agaagagggc actctgaacg
     4501 tcgttcagct ggtcgcggga gatggtaccc caatgactcc gcatgcagtg cgtgatcgca
     4561 ttcaggctta ccgtgacgaa caggcacatt tactggatga gctgcatcgc agcgcgcccg
     4621 caagtgccgc cgctttagaa aataacctgc gcgtagccat agatttgtat acagttgatg
     4681 atttgcagtt tgaccttccg gacgccgaac aagccgttgg ctaataagcc tcgttgctct
     4741 tcataaatca tccctatgac cccttcaaaa aaaaaaaaca ccctaacggg tgtttttttt
     4801 tttttggtct gcctctcgct gacggtcaca atgcgcttac gaaagtcttc ctgtactagt
     4861 agcggccgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta
     4921 aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg
     4981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatgac agtaagacgg
     5041 gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg aatgacctgt
     5101 cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg ctgaacagca
     5161 aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa gcccgtgacg
     5221 ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc
     5281 catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc acgaaaaaga
     5341 cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt tgcccgagct
     5401 tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag gagcaaacag
     5461 cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt tatacacagg
     5521 gctgggatct attcttttta tcttttttta ttctttcttt attctataaa ttataaccac
     5581 ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg gtctagcaga
     5641 atttacaagt tttccagcaa aggtctagca gaatttacag atacccacaa ctcaaaggaa
     5701 aaggactagt aattatcatt gactagccca tctcaattgg tatagtgatt aaaatcacct
     5761 agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac tagcgattag
     5821 tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg cactactcaa
     5881 ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt ataaccaata
     5941 cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta aagcaaccag
     6001 agagctgatg acgagaactg tggaaatcag gaatcctttg gttaaaggct tttggatttt
     6061 ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt ttagtgaaga
     6121 gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg aacatgttaa
     6181 gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag aactaacaca
     6241 aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt tcatgttaat
     6301 gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga aaccaataag
     6361 taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag gccgcccgac
     6421 tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa ccgaacttga
     6481 gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat cagattccta
     6541 cctacataac ggactaagaa aaacactaca cgatgcttta actgcaaaaa ttcagctcac
     6601 cagttttgag gcaaaatttt tgagtgacat gcaaagtaag tatgatctca atggttcgtt
     6661 ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta aatacggaag
     6721 gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta acaaacaaaa
     6781 gtagaacaac tgttcaccgt tacatatcaa agggaaaact gtccatatgc acagatgaaa
     6841 acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca ttcaaagctg
     6901 ttcaccatga acagatcgac aatgtaacag atgaacagca tgtaacacct aatagaacag
     6961 gtgaaaccag taaaacaaag caactagaac atgaaattga acacctgaga caacttgtta
     7021 cagctcaaca gtcacacata gacagcctga aacaggcgat gctgcttatc gaatcaaagc
     7081 tgccgacaac acgggagcca gtgacgcctc ccgtggggaa aaaatcatgg caattctgga
     7141 agaaatagcg ctttcagccg gcaaaccggc tgaagccgga tctgcgattc tgataacaaa
     7201 ctagcaacac cagaacagcc cgtttgcggg cagcaaaacc cgtaccgatt atcaaaaagg
     7261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7441 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     7501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8041 atcttaccgc tgttgagatc cagttcgata taacccactc gtgcacccaa ctgatcttca
     8101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8281 aaaaataaac aaataggggt tccgcg
//
