LOCUS       sequence-413673-        9714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of sclareol in E. coli MEV20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9714)
  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 9714)
  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: pLRD11
FEATURES             Location/Qualifiers
     source          1..9714
                     /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        4375..4393
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4394..4418
                     /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    6262..6282
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      6283..6340
                     /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(6364..6381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      6915..7137
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7185..8135
                     /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(8749..9540,9541..9609))
                     /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     9372..9391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9610..9714)
                     /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 agagctctat ctaataagga gaactataat
     1921 gaatcgtgaa ttcccgccga gcttttggaa caatgacatt ataaacagca taacagcgag
     1981 ccataaagta cagactggtg atcgcaagcg cattcagacg ctgatttccg aaattaaaaa
     2041 tgtgtttaat agcatggggg acggggaaac aagcccgtcg gcatacgata cggcatgggt
     2101 ggcgcgcatt cctgctgttg atggttccga acagccgcag ttcccgcaga cgctggagtg
     2161 gattttacag aaccaactta aagatggatc gtggggcgag gaattttatt tcttagcgta
     2221 cgatcggtta cttgcaacgc tggcgtgcat cattaccctg accatctggc gcaccggcaa
     2281 cgtccaatta cacaaaggga ttgagttctt tcgcaaacag gtagtacgga tggacgacga
     2341 ggcggacaat catcgcccca gcggctttga aattgtcttt ccggcgatgc tgaacgaagc
     2401 aaaatctctg ggtctggacc tgccttatga acttccgttc atcgaacaga tggtgaagaa
     2461 acgcgaagcg aaattgaaga tgattacgac caatgtactc tataccattc agaccacgct
     2521 gttatacagt ttggaaggcc tccacgagat cgtggacttt gataaaatta ttaaactcca
     2581 atccaaggat gggagctttc tcggtagtcc ggcgtctacc gcggctgtgt tcatgcaaac
     2641 cggtaatacc aaatgccttg aatttctgga gttcgttctg cgaaagtttc gtaatcatgt
     2701 gccttcagat tatccgctgg atctgttcga gcgcctctgg gtggtggata ccgttgaacg
     2761 tctgggtatt gaccgtcact ttaagaaaga aatcaaagac gcgctggact atgtctattc
     2821 ttgttgggat gaacgaggaa ttggctgggc gaaggattcc cctatcgcag atattgatga
     2881 tactgccatg ggtctgcgca tcctgcgtct gcacgggtat aatgttagtc cggatgttct
     2941 gaaaaccttt aaggacgaga acggtgaatt cttttgcttt atgggccaga cgcaaagagg
     3001 tgtgaccgat atgctgaacg tgtaccgttg cagtcaagtg gcattccccg gggaaaccat
     3061 catggaggaa gcgaaattat gcacagaacg ttacttgcgc aacgctttag aaaacgctga
     3121 tgccttcgat aagtgggcga tcaagaaaaa cattcgcgga gaggttgaat atgccttaaa
     3181 atatccttgg catcgaagca tgccacggct ggaagtccgt tcgtacattg gaaattatgg
     3241 tccaaacgat gtttggctgg gtaaaagtct gtatatgatg ccgtatattt ccaacgagaa
     3301 atacctggaa cttgcgaaac ttgatttcaa tagcgtccag tctctgcacc aggaagaaat
     3361 ccgcgaatta gtgcgctggt gtaaatcatc aggatttacg gaactgaagt ttacgcgtga
     3421 ccgcgttgtg gaaacgtact ttgccgtggc atcaagcatg tttgaaccag aattctccac
     3481 ttgccgtgct gtgtatacga aaatcagcgt gctccttgta atcctggctg acctgtatga
     3541 tggttacggc tccccggatg agattaaact gttctctgag gcggtgaaac gttgggacct
     3601 gagcctgctg gaacaaatgc cggatcacat gaaaatttgc ttcctgggcc tttacaacac
     3661 cgtaaatgag gtggcggaag aggggcgtaa gacccagggt catgatgtac ttggttacat
     3721 ccggaatctg tgggagattc aactggcagc atttacccgt gaagcagaat ggtcccaggg
     3781 aaaatatgtg ccatcgtttg acgaatatat tgagaacgcg caagtaagca ttggtgtggc
     3841 tacaatcctg ctgattacca tcttgtttac tgaagaagat gacatcctga gccacattga
     3901 ttatggttcc aaatttcttc gactggcgag tctcactgcg cgtttggcga atgacattaa
     3961 aacttatcaa gaagaacggg cgcacggcga agtggttagc gcaattcaat gctacatgaa
     4021 agatcgcccg gaaatcacgg aagaagaagc cctgaaatat gtgtacggcc gcatggtcaa
     4081 cgatctggcc gaactgaata gcgaatacct gaaaagtaac gaaatgcccc agaactgcaa
     4141 gcgcttagta ttcgacaccg cgcgcgtcgc acagctgttt accatggagg gcgacgggct
     4201 cacgtattcg gataccatgg agatcaagga acacatcaag aaatgcttat tcgaaccggc
     4261 aacttaataa taattatgcc ttccgctaat caactcggta ccaaattcca gaaaagagac
     4321 gctttcgagc gtcttttttc gttttggtcc gtggacaagc acagcatagc caagtaatac
     4381 gactcactat aggggaattg tgagcggata acaattcccc tctaggtccg aagatactta
     4441 gggtgtctca aggtgcgtac cttgactgat gagtccgaaa ggacgaaaca cccctctaca
     4501 aataattttg tttaagatat aatcaaaaat tacaaataaa ctaggagtcg ttaaatggct
     4561 aaaatgaaag aaaactttaa acgtgaggac gataagtttc caactactac cacgctccgg
     4621 agcgaagaca tcccctcaaa tttgtgtatc atcgatactc tgcaacgact cggagttgat
     4681 cagttcttcc agtatgaaat taacacgata ctggacaata catttcgcct gtggcaggaa
     4741 aaacacaagg tcatttatgg aaacgtgacc acccatgcga tggcctttcg cttgctgcgt
     4801 gtcaaaggct acgaagtttc ctcggaagag ttggctccgt acggtaacca agaagcggtg
     4861 tcccagcaga caaatgatct gcccatgatt attgagttat accgtgcagc caacgaacga
     4921 atttatgagg aagaacgttc acttgagaaa attctggctt ggacaaccat cttcctgaat
     4981 aagcaggttc aagataactc cattccggat aagaaactgc acaaactggt tgagttttac
     5041 ctgcgtaatt acaaggggat caccatccgc ctgggcgcca gacgtaatct ggaattatac
     5101 gatatgacat actatcaagc tctcaaaagc acaaaccgtt tctcgaattt atgcaatgaa
     5161 gattttcttg tgttcgccaa acaggatttc gatatacatg aggcacagaa tcagaaaggc
     5221 ctgcagcagc tgcagcgctg gtatgctgac tgccgtttag ataccctgaa tttcggtcgg
     5281 gatgtcgtga ttgttgctaa ctacctggcc agtcttatta tcggcgatca cgcgtttgac
     5341 tacgttcgct tagcgtttgc caagactagt gtccttgtta ccatcatgga tgatttcttt
     5401 gattgccacg gcagctccca ggaatgcgac aaaatcatcg agctcgtcaa agagtggaag
     5461 gaaaacccag atgcagagta tggtagtgaa gaactggaaa tacttttcat ggcactgtat
     5521 aacaccgtca atgaattagc ggaacgcgcg cgcgtggaac aagggcggag cgtgaaagag
     5581 tttttagtca aactctgggt ggagattctt agcgccttca aaatcgagct ggacacttgg
     5641 tcgaatggga cgcagcaaag tttcgatgaa tacatcagct caagctggct ttccaacggt
     5701 agccgtctga caggcctcct taccatgcag ttcgtgggag ttaaactctc agacgagatg
     5761 ctgatgagtg aggaatgcac cgacttagcc cgccatgtat gtatggtcgg tcggcttctg
     5821 aacgacgtct gctctagtga acgagagcgc gaagaaaaca ttgcgggtaa gtcctattca
     5881 attctgctcg ctaccgaaaa agatggccgt aaagtgtctg aagatgaagc gatcgctgaa
     5941 attaatgaga tggttgaata tcactggcgc aaagtgttgc aaatcgtgta taaaaaggag
     6001 agcattctgc cgcgtcgctg caaagacgtg ttcctggaaa tggccaaagg tacattctac
     6061 gcctatggca taaatgatga actcacgagt ccacagcaga gtaaagagga catgaaaagt
     6121 tttgttttct aataagcctc gttgctcttc ataaatcatc cctatgaccc cttcaaaaaa
     6181 aaaaaacacc ctaacgggtg tttttttttt tttggtctgc ctctcgctga cggtcacaat
     6241 gcgcttacga aagtcttcct gtactagtag cggccgctgc agtccggcaa aaaagggcaa
     6301 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc
     6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6421 ctcaaaggcg gtaatgacag taagacgggt aagcctgttg atgataccgc tgccttactg
     6481 ggtgcattag ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa
     6541 atcagagggc aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc
     6601 cataaaacgc cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc
     6661 atgaatccat aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag
     6721 cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa
     6781 aaggaatatt cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt
     6841 aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa
     6901 ctgactaaag tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt
     6961 ctttctttat tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc
     7021 tagcggaatt tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga
     7081 atttacagat acccacaact caaaggaaaa ggactagtaa ttatcattga ctagcccatc
     7141 tcaattggta tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg
     7201 ttttcaaagc aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc
     7261 taattttatg ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac
     7321 ggacggtatc gttcacttat aaccaatacg ctcagatgat gaacatcagt agggaaaatg
     7381 cttatggtgt attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga
     7441 atcctttggt taaaggcttt tggattttcc agtggacaaa ctatgccaag ttctcaagcg
     7501 aaaaattaga attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat
     7561 tcataaaata taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt
     7621 atgagtggtt attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta
     7681 gccttgatga atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc
     7741 ttaaccaatg ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat
     7801 tggtggttga taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata
     7861 gacaaatgga tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa
     7921 taccaacaac cattacatca gattcctacc tacataacgg actaagaaaa acactacacg
     7981 atgctttaac tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc
     8041 aaagtaagta tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaaccacac
     8101 tagagaacat actggctaaa tacggaagga tctgaggttc ttatggctct tgtatctatc
     8161 agtgaagcat caagactaac aaacaaaagt agaacaactg ttcaccgtta catatcaaag
     8221 ggaaaactgt ccatatgcac agatgaaaac ggtgtaaaaa agatagatac atcagagctt
     8281 ttacgagttt ttggtgcatt caaagctgtt caccatgaac agatcgacaa tgtaacagat
     8341 gaacagcatg taacacctaa tagaacaggt gaaaccagta aaacaaagca actagaacat
     8401 gaaattgaac acctgagaca acttgttaca gctcaacagt cacacataga cagcctgaaa
     8461 caggcgatgc tgcttatcga atcaaagctg ccgacaacac gggagccagt gacgcctccc
     8521 gtggggaaaa aatcatggca attctggaag aaatagcgct ttcagccggc aaaccggctg
     8581 aagccggatc tgcgattctg ataacaaact agcaacacca gaacagcccg tttgcgggca
     8641 gcaaaacccg taccgattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8881 gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     9001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatata
     9481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcg
//
