LOCUS       sequence-431886-        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express amorphadiene synthase (ADS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  AUTHORS   Baker JJ, Shi J, Wang S, Mujica EM, Bianco S, Capponi S, Dueber JE
  TITLE     ML-enhanced peroxisome capacity enables compartmentalization of 
            multienzyme pathway.
  JOURNAL   Nat Chem Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2.
  PUBMED    39402374
REFERENCE   2  (bases 1 to 6829)
  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: "Nat Chem 
            Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2."
COMMENT     Sequence Label: pJJB1734
FEATURES             Location/Qualifiers
     source          1..6829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..680
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     550..573
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     RBS             2126..2134
                     /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)"
     terminator      2340..2527
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2395..2419)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2812..2999
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3000..3662
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      3877..4859
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4917..5732)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4917..5732)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5640..5659
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5724..5743
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5957..6545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6446..6465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt
       61 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
      121 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
      181 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
      241 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
      301 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
      361 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
      421 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
      481 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
      541 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
      601 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata
      661 aacacacata aacaaacaaa agatctatgt cattgactga ggaaaaacct atccgaccga
      721 ttgcaaattt tcctccatcc atatggggtg atcagttctt gatttacgaa aaacaagtcg
      781 agcaaggtgt tgaacaaatt gtaaatgatt tgaaaaaaga ggtgagacag ttattgaagg
      841 aggcattaga tattccaatg aaacacgcta atttattaaa gcttatcgac gaaatccaac
      901 gattaggtat tccttatcat ttcgagagag aaattgatca cgctttgcaa tgtatatacg
      961 aaacatatgg agacaattgg aatggtgata ggagtagtct gtggttcaga ctaatgagaa
     1021 aacaaggcta ctatgtcaca tgtgatgtat ttaataatta taaggataaa aatggtgcat
     1081 ttaagcagag cctagcgaat gatgttgaag gactgttgga attatatgag gctacctcta
     1141 tgagagtgcc tggtgagatt atattagaag atgccttggg cttcacgaga tcacgtttat
     1201 cgattatgac gaaggacgct ttttctacga acccagcact ttttacagaa attcagagag
     1261 ctttgaagca gccactttgg aaaagactac ctcgtataga ggcagctcag tatatccctt
     1321 tttaccaaca gcaagatagc cataacaaaa ctttacttaa attggctaaa cttgaattca
     1381 accttcttca atcattacac aaagaagaac tgagccatgt gtgtaagtgg tggaaggcct
     1441 tcgacatcaa aaaaaacgca ccatgtttga gagacaggat tgtagaatgc tacttttggg
     1501 gtttgggctc tggttacgaa ccacaatatt caagggccag agttttcttt accaaggctg
     1561 tcgcagtaat aactctaatt gacgatactt acgacgccta tggcacttat gaagaattga
     1621 agatatttac agaagcagtt gaacgctggt ctattacttg tttagataca ttaccagaat
     1681 atatgaaacc gatatacaaa ctatttatgg acacttatac agaaatggag gaatttctcg
     1741 ccaaagaagg aaggaccgat ttatttaatt gcggtaaaga attcgtcaag gaatttgtta
     1801 gaaatctcat ggttgaggcg aaatgggcta acgaaggcca tattccaacg accgaggagc
     1861 atgaccctgt tgtcattata actggtgggg ctaatttact tactactacc tgctacttag
     1921 ggatgtccga tatattcaca aaggaatcag ttgagtgggc tgtttctgca cccccactgt
     1981 ttcggtacag tggaatcctg ggacgtaggt tgaatgactt gatgactcat aaagccgaac
     2041 aagaaagaaa gcactcgtcc tcgtctttag aatcctatat gaaagaatac aacgtcaacg
     2101 aagaatatgc ccaaacattg atttataagg aggtagaaga tgtatggaaa gatatcaacc
     2161 gtgaatacct cacaaccaag aatatcccca gaccgctact aatggctgtg atatacttgt
     2221 gtcaattttt ggaagttcaa tatgcgggta aggataactt caccaggatg ggggatgagt
     2281 ataaacatct gatcaagtct ctcctagtgt atcccatgag tattggatcc taactcgagg
     2341 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     2401 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     2461 tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct
     2521 ctaccggcat gccgagcaaa tgcctgcaaa tcgctcccca tttcgctgga aatctgctcg
     2581 tcagtggtgc tcacactgac gaatcatgta cagatcatac cgatgactgc ctggcgactc
     2641 acaactaagc aagacagccg gaaccagcgc cggcgaacac cactgcatat atggcatatc
     2701 acaacagtcc acgtctcaag cagttacaga gatgttacga accactagtg cactgcagta
     2761 caaacacagt cctttcccgc aattttcttt ttctattact cttggcctcc tctagtacac
     2821 tctatatttt tttatgcctc ggtaatgatt ttcatttttt tttttccacc tagcggatga
     2881 ctcttttttt ttcttagcga ttggcattat cacataatga attatacatt atataaagta
     2941 atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg aaggcaaaga
     3001 tgacagagca gaaagcccta gtaaagcgta ttacaaatga aaccaagatt cagattgcga
     3061 tctctttaaa gggtggtccc ctagcgatag agcactcgat cttcccagaa aaagaggcag
     3121 aagcagtagc agaacaggcc acacaatcgc aagtgattaa cgtccacaca ggtatagggt
     3181 ttctggacca tatgatacat gctctggcca agcattccgg ctggtcgcta atcgttgagt
     3241 gcattggtga cttacacata gacgaccatc acaccactga agactgcggg attgctctcg
     3301 gtcaagcttt taaagaggcc ctaggggccg tgcgtggagt aaaaaggttt ggatcaggat
     3361 ttgcgccttt ggatgaggca ctttccagag cggtggttga tctttcgaac aggccgtacg
     3421 cagttgtcga acttggtttg caaagggaga aagtaggtga tctctcttgc gagatgatcc
     3481 cgcattttct tgaaagcttt gcagaggcta gcagaattac cctccacgtt gattgtctgc
     3541 gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa ggctcttgcg gttgccataa
     3601 gagaagccac ctcgcccaat ggtaccaacg atgttccctc caccaaaggt gttcttatgt
     3661 agtgacaccg attatttaaa gctgctgcat acgatatata tacatgtgta tatatgtata
     3721 cctatgaatg tcagtaagta tgtatacgaa cagtatgata ctgaagatga caaggtaatg
     3781 catcattcta tacgtgtcat tctgaacgag gcgcgctttc cttttttctt tttgcttttt
     3841 cttttttttt ctcttgaact cgacggatca tagagtaacg aagcatctgt gcttcatttt
     3901 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3961 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     4021 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     4081 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     4141 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     4201 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     4261 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     4321 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     4381 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     4441 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     4501 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4561 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4621 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4681 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4741 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4801 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4861 tagagaatag gaacttcccg agcggccgcg tgttacaacc aattaaccaa ttctgattag
     4921 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     4981 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     5041 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     5101 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     5161 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     5221 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     5281 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     5341 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     5401 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     5461 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     5521 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     5581 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     5641 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg
     5701 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     5761 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     5821 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     5881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6601 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gcccctgaat
     6661 tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc
     6721 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
     6781 cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacg
//
