LOCUS       sequence-413689-        7850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of cis-abienol in E. coli MEV20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  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 7850)
  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: pLRD25
FEATURES             Location/Qualifiers
     source          1..7850
                     /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)."
     misc_feature    4398..4418
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4419..4476
                     /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(4500..4517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5051..5273
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5321..6271
                     /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(6885..7676,7677..7745))
                     /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     7508..7527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7746..7850)
                     /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 gggcaagcaa gtaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     1801 aggtccgaag atacttaggg tgtctcaagg tgcgtacctt gactgatgag tccgaaagga
     1861 cgaaacaccc ctctacaaat aattttgttt aagaggtcta tctaataagg agaactataa
     1921 tgaatcgtga attcccgccg agcttttgga acaatgacat tataaacagc ataacagcga
     1981 gccataaagt acagactggt gatcgcaagc gcattcagac gctgatttcc gaaattaaaa
     2041 atgtgtttaa tagcatgggg gacggggaaa caagcccgtc ggcatacgat acggcatggg
     2101 tggcgcgcat tcctgctgtt gatggttccg aacagccgca gttcccgcag acgctggagt
     2161 ggattttaca gaaccaactt aaagatggat cgtggggcga ggaattttat ttcttagcgt
     2221 acgatcggtt acttgcaacg ctggcgtgca tcattaccct gaccatctgg cgcaccggca
     2281 acgtccaatt acacaaaggg attgagttct ttcgcaaaca ggtagtacgg atggacgacg
     2341 aggcggacaa tcatcgcccc agcggctttg aaattgtctt tccggcgatg ctgaacgaag
     2401 caaaatctct gggtctggac ctgccttatg aacttccgtt catcgaacag atggtgaaga
     2461 aacgcgaagc gaaattgaag atgattacga ccaatgtact ctataccatt cagaccacgc
     2521 tgttatacag tttggaaggc ctccacgaga tcgtggactt tgataaaatt attaaactcc
     2581 aatccaagga tgggagcttt ctcggtagtc cggcgtctac cgcggctgtg ttcatgcaaa
     2641 ccggtaatac caaatgcctt gaatttctgg agttcgttct gcgaaagttt cgtaatcatg
     2701 tgccttcaga ttatccgctg gatctgttcg agcgcctctg ggtggtggat accgttgaac
     2761 gtctgggtat tgaccgtcac tttaagaaag aaatcaaaga cgcgctggac tatgtctatt
     2821 cttgttggga tgaacgagga attggctggg cgaaggattc ccctatcgca gatattgatg
     2881 atactgccat gggtctgcgc atcctgcgtc tgcacgggta taatgttagt ccggatgttc
     2941 tgaaaacctt taaggacgag aacggtgaat tcttttgctt tatgggccag acgcaaagag
     3001 gtgtgaccga tatgctgaac gtgtaccgtt gcagtcaagt ggcattcccc ggggaaacca
     3061 tcatggagga agcgaaatta tgcacagaac gttacttgcg caacgcttta gaaaacgctg
     3121 atgccttcga taagtgggcg atcaagaaaa acattcgcgg agaggttgaa tatgccttaa
     3181 aatatccttg gcatcgaagc atgccacggc tggaagtccg ttcgtacatt ggaaattatg
     3241 gtccaaacga tgtttggctg ggtaaaagtc tgtatatgat gccgtatatt tccaacgaga
     3301 aatacctgga acttgcgaaa cttgatttca atagcgtcca gtctctgcac caggaagaaa
     3361 tccgcgaatt agtgcgctgg tgtaaatcat caggatttac ggaactgaag tttacgcgtg
     3421 accgcgttgt ggaaacgtac tttgccgtgg catcaagcat gtttgaacca gaattctcca
     3481 cttgccgtgc tgtgtatacg aaaatcagcg tgctccttgt aatcctggat gacctgtatg
     3541 atggttacgg ctccccggat gagattaaac tgttctctga ggcggtgaaa cgttgggacc
     3601 tgagcctgct ggaacaaatg ccggatcaca tgaaaatttg cttcctgggc ctttacaaca
     3661 ccgtaaatga ggtggcggaa gaggggcgta agacccaggg tcatgatgta cttggttaca
     3721 tccggaatct gtgggagatt caactggcag catttacccg tgaagcagaa tggtcccagg
     3781 gaaaatatgt gccatcgttt gacgaatata ttgagaacgc gcaagtaagc attggtgtgg
     3841 ctacaatcct gctgattacc atcttgttta ctgaagaaga tgacatcctg agccacattg
     3901 attatggttc caaatttctt cgactggcga gtctcactgc gcgtttggcg aatgacatta
     3961 aaacttatca agaagaacgg gcgcacggcg aagtggttag cgcaattcaa tgctacatga
     4021 aagatcgccc ggaaatcacg gaagaagaag ccctgaaata tgtgtacggc cgcatggtca
     4081 acgatctggc cgaactgaat agcgaatacc tgaaaagtaa cgaaatgccc cagaactgca
     4141 agcgcttagt attcgacacc gcgcgcgtcg cacagctgtt taccatggag ggcgacgggc
     4201 tcacgtattc ggataccatg gagatcaagg aacacatcaa gaaatgctta ttcgaaccgg
     4261 caacttaata agcctcgttg ctcttcataa atcatcccta tgaccccttc aaaaaaaaaa
     4321 aacaccctaa cgggtgtttt tttttttttg gtctgcctct cgctgacggt cacaatgcgc
     4381 ttacgaaagt cttcctgtac tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg
     4441 tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc
     4501 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4561 aaggcggtaa tgacagtaag acgggtaagc ctgttgatga taccgctgcc ttactgggtg
     4621 cattagccag tctgaatgac ctgtcacggg ataatccgaa gtggtcagac tggaaaatca
     4681 gagggcagga actgctgaac agcaaaaagt cagatagcac cacatagcag acccgccata
     4741 aaacgccctg agaagcccgt gacgggcttt tcttgtatta tgggtagttt ccttgcatga
     4801 atccataaaa ggcgcctgta gtgccattta cccccattca ctgccagagc cgtgagcgca
     4861 gcgaactgaa tgtcacgaaa aagacagcga ctcaggtgcc tgatggtcgg agacaaaagg
     4921 aatattcagc gatttgcccg agcttgcgag ggtgctactt aagcctttag ggttttaagg
     4981 tctgttttgt agaggagcaa acagcgtttg cgacatcctt ttgtaatact gcggaactga
     5041 ctaaagtagt gagttataca cagggctggg atctattctt tttatctttt tttattcttt
     5101 ctttattcta taaattataa ccacttgaat ataaacaaaa aaaacacaca aaggtctagc
     5161 ggaatttaca gagggtctag cagaatttac aagttttcca gcaaaggtct agcagaattt
     5221 acagataccc acaactcaaa ggaaaaggac tagtaattat cattgactag cccatctcaa
     5281 ttggtatagt gattaaaatc acctagacca attgagatgt atgtctgaat tagttgtttt
     5341 caaagcaaat gaactagcga ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat
     5401 tttatgctgt gtggcactac tcaaccccac gattgaaaac cctacaagga aagaacggac
     5461 ggtatcgttc acttataacc aatacgctca gatgatgaac atcagtaggg aaaatgctta
     5521 tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc
     5581 tttggttaaa ggcttttgga ttttccagtg gacaaactat gccaagttct caagcgaaaa
     5641 attagaatta gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat
     5701 aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga
     5761 gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct
     5821 tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa
     5881 ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt
     5941 ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca
     6001 aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc
     6061 aacaaccatt acatcagatt cctacctaca taacggacta agaaaaacac tacacgatgc
     6121 tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag
     6181 taagtatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga
     6241 gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg
     6301 aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata tcaaagggaa
     6361 aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac
     6421 gagtttttgg tgcattcaaa gctgttcacc atgaacagat cgacaatgta acagatgaac
     6481 agcatgtaac acctaataga acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa
     6541 ttgaacacct gagacaactt gttacagctc aacagtcaca catagacagc ctgaaacagg
     6601 cgatgctgct tatcgaatca aagctgccga caacacggga gccagtgacg cctcccgtgg
     6661 ggaaaaaatc atggcaattc tggaagaaat agcgctttca gccggcaaac cggctgaagc
     6721 cggatctgcg attctgataa caaactagca acaccagaac agcccgtttg cgggcagcaa
     6781 aacccgtacc gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7021 ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatataaccc
     7621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
//
