LOCUS       sequence-360446-        9407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing ERG20 (GAL10 promoter) and 
            fusion protein deadFPPS-AcNES1 (GAL7 promoter). deadFPPS is 
            ERG20(K197G-K254A), an inactive mutant of ERG20..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9407)
  AUTHORS   Cheah LC, Liu L, Stark T, Plan MR, Peng B, Lu Z, Schenk G, Sainsbury
            F, Vickers CE
  TITLE     Metabolic flux enhancement from the translational fusion of terpene 
            synthases is linked to terpene synthase accumulation.
  JOURNAL   Metab Eng. 2023 May;77:143-151. doi: 10.1016/j.ymben.2023.03.012. 
            Epub 2023 Mar 28.
  PUBMED    36990382
REFERENCE   2  (bases 1 to 9407)
  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: "Metab 
            Eng."; date: "2023-05"; volume: "77:143-151. doi"; pages: " 
            10.1016/j.ymben.2023.03.012. Epub 2023 Mar 28"
COMMENT     Sequence Label: Coexpressed Free deadFPPS-NES
FEATURES             Location/Qualifiers
     source          1..9407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    300..417
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(6520..6537)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6520..6536)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6529..6551)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6745..6764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6864..6886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6924..6942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7010..7114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7115..7183,7184..7975)
                     /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     complement(7333..7352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8146..8734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8635..8654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8888..8905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9022..9043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9058..9063,9064..9081,9082..9088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9096..9112
                     /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)."
     primer_bind     9101..9123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9120..9136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9120..9136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(9383..9407,1..417))
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(join(9389..9407,1..5))
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 atattaacaa ttttttgttg atacttttat gacatttgaa taagaagtaa tacaaaccga
       61 aaatgttgaa agtattagtt aaagtggtta tgcagctttt gcatttatat atctgttaat
      121 agatcaaaaa tcatcgcttc gctgattaat taccccagaa ataaggctaa aaaactaatc
      181 gcattattat cctatggttg ttaatttgat tcgttgattt gaaggtttgt ggggccaggt
      241 tactgccaat ttttcctctt cataaccata aaagctagta ttgtagaatc tttattgttc
      301 ggagcagtgc ggcgcgaggc acatctgcgt ttcaggaacg cgaccggtga agaccaggac
      361 gcacggagga gagtcttccg tcggagggct gtcgcccgct cggcggcttc taatccgtac
      421 ttcaatatag caatgagcag ttaagcgtat tactgaaagt tccaaagaga aggttttttt
      481 aggctaagat aatggggctc tttacatttc cacaacatat aagtaagatt agatatggat
      541 atgtatatgg tggtattgcc atgtaatatg attattaaac ttctttgcgt ccatccaaaa
      601 aaaaaagtaa gaatttttga aaattcaata taaggatcca aaatggcttc agaaaaagaa
      661 attaggagag agagattctt gaacgttttc cctaaattag tagaggaatt gaacgcatcg
      721 cttttggctt acggtatgcc taaggaagca tgtgactggt atgcccactc attgaactac
      781 aacactccag gcggtaagtt aaatagaggt ttgtccgttg tggacacgta tgctattctc
      841 tccaacaaga ccgttgaaca attggggcaa gaagaatacg aaaaggttgc tattctaggt
      901 tggtgcattg agttgttgca ggcttacttc ttggtcgccg atgatatgat ggacaagtcc
      961 attaccagaa gaggccaacc atgttggtac aaggttcctg aagttgggga aattgccatc
     1021 aatgacgcat tcatgttaga ggctgctatc tacaagcttt tgaaatctca cttcagaaac
     1081 gaaaaatact acatagatat caccgaattg ttccatgaag tcaccttcca aaccgaattg
     1141 ggccaattga tggacttaat cactgcacct gaagacaaag tcgacttgag taagttctcc
     1201 ctaaagaagc actccttcat agttactttc aagactgctt actattcttt ctacttgcct
     1261 gtcgcattgg ctatgtacgt tgccggtatc acagatgaaa aggatttgaa acaagccaga
     1321 gatgtcttga ttccattggg tgaatatttc caaattcaag atgactactt agactgcttc
     1381 ggtaccccag aacagatcgg taagatcggt acagatatcc aagataacaa atgttcttgg
     1441 gtaatcaaca aggcattaga acttgcttcc gcagaacaaa gaaagacttt agacgaaaat
     1501 tacggtaaga aggactcagt cgcagaagcc aaatgcaaaa agattttcaa tgacttgaaa
     1561 atcgaccagt tataccacga atatgaagag tctgttgcca aggatttgaa ggccaagatc
     1621 tcccaagtcg acgagtctcg tggcttcaaa gccgacgtct taactgcgtt tttgaacaaa
     1681 gtttacaaga gaagcaaata aagatctttt gccagcttac tatccttctt gaaaatatgc
     1741 actctatatc ttttagttct taattgcaac acatagattt gctgtataac gaattttatg
     1801 ctatttttta aatttggagt tcagtgataa aagtgtcaca gcgaatttcc tcacatgtag
     1861 ggaccgaatt gtttacaagt tctctgtacc accatggaga catcaaaaat tgaaaatcta
     1921 tggaaagata tggacggtag caacaagaat atagcacgag ccgcggagtt catttcgtta
     1981 cttttgatat cactcacaac tattgcgaag cgcttcagtg aaaaaatcat aaggaaaagt
     2041 tgtaaatatt attggtagta ttcgtttggt aaagtagagg gggtaatttt tcccctttat
     2101 tttgttcata cattcttaaa ttgctttgcc tctccttttg gaaagctata cttcggagca
     2161 ctgttgagcg aaggctcatt agatatattt tctgtcattt tccttaaccc aaaaataagg
     2221 gaaagggtcc aaaaagcgct cggacaactg ttgaccgtga tccgaaggac tggctataca
     2281 gtgttcacaa aatagccaag ctgaaaataa tgtgtagcta tgttcagtta gtttggctag
     2341 caaagatata aaagcaggtc ggaaatattt atgggcatta ttatgcagag catcaacatg
     2401 ataaaaaaaa acagttgaat attccctcaa aagagctcaa aatggcttca gaaaaagaaa
     2461 ttaggagaga gagattcttg aacgttttcc ctaaattagt agaggaattg aacgcatcgc
     2521 ttttggctta cggtatgcct aaggaagcat gtgactggta tgcccactca ttgaactaca
     2581 acactccagg cggtaagtta aatagaggtt tgtccgttgt ggacacgtat gctattctct
     2641 ccaacaagac cgttgaacaa ttggggcaag aagaatacga aaaggttgct attctaggtt
     2701 ggtgcattga gttgttgcag gcttacttct tggtcgccga tgatatgatg gacaagtcca
     2761 ttaccagaag aggccaacca tgttggtaca aggttcctga agttggggaa attgccatca
     2821 atgacgcatt catgttagag gctgctatct acaagctttt gaaatctcac ttcagaaacg
     2881 aaaaatacta catagatatc accgaattgt tccatgaagt caccttccaa accgaattgg
     2941 gccaattgat ggacttaatc actgcacctg aagacaaagt cgacttgagt aagttctccc
     3001 taaagaagca ctccttcata gttactttcg gtactgctta ctattctttc tacttgcctg
     3061 tcgcattggc tatgtacgtt gccggtatca cagatgaaaa ggatttgaaa caagccagag
     3121 atgtcttgat tccattgggt gaatatttcc aaattcaaga tgactactta gactgcttcg
     3181 gtaccccaga acagatcgat gctatcggta cagatatcca agataacaaa tgttcttggg
     3241 taatcaacaa ggcattagaa cttgcttccg cagaacaaag aaagacttta gacgaaaatt
     3301 acggtaagaa ggactcagtc gcagaagcca aatgcaaaaa gattttcaat gacttgaaaa
     3361 tcgaccagtt ataccacgaa tatgaagagt ctgttgccaa ggatttgaag gccaagatct
     3421 cccaagtcga cgagtctcgt ggcttcaaag ccgacgtctt aactgcgttt ttgaacaaag
     3481 tttacaagag aagcaaaaga tctgctggtg gaggaggtac tggtggtgct gagctcaaaa
     3541 tggctaccgc agcaggtcct atcgcaacta acaactcccc acaaaactcc aacgcttaca
     3601 gaactccaat cgctccttcc gtaccaatta ctcataaatg gtctatagct gaagatttga
     3661 catgtatttc caatcctagt aagcacaata accctcaaac tggttacaga tcattttctg
     3721 acgaattata cgttaagtac gaagaaaagt tggaagatgt tagaaaagca ttaagagaag
     3781 ttgaagaaaa ccctttggaa ggtttagtta tgatagacgc tttgcaaaga ttgggtatcg
     3841 attaccattt cagaggtgaa attggtgcat tcttgcaaaa gcaacaaatc atatcttcaa
     3901 ctccagatgg ttaccctgaa catggtttgt acgaagtttc aacattgttt agattcttaa
     3961 gacaagaagg tcacaatgtt accgctgacg tctttaataa cttcaaggat aaggaaggta
     4021 gattcagatc agaattgtca acagatatta gaggtttgat gtccttatac gaagcaagtc
     4081 aattgagaat agaaggtgaa gacatcttag atcaagctgc tgatttctcc agtcaattgt
     4141 taggtagatg gacaaaagat cctaatcatc acgaagccag attggtttct aacactttaa
     4201 cacatccata ccacaagtca ttggctacct ttatgggtca aaaattgtcc tacatgaact
     4261 gcaagggtcc aaactgggac ggtgtcgata atttgcaaga attagctaag atggatttga
     4321 ctatcgtaca aagtatccat caaaaggaag tattccaagt ttctcaatgg tggaaggata
     4381 caggtttagc caatgaattg aaattggcta gaaaccaacc attgaagtgg tatatgtggc
     4441 ctatggccgc tttaaccgat ccaagattca gtgaagaaag agttgaatta actaagccta
     4501 tttcttttat atacatcata gatgacatct tcgacgtcta tggtaccatt gaagaattga
     4561 ccttgtttac tgatgcagtt aatagatggg aattgtctgc cgtcgaacaa ttaccagact
     4621 acatgaaagt atgtttcaag gctttgtacg atgttaccaa cgaaatcgca tacaaaatct
     4681 ataaaaagca tggtcaaaac cctattgatt ccttgcaaaa gacttgggct agtttgtgca
     4741 atgcattttt agttgaagca aagtggttcg cctctggtca cttgccaaat gcagaagaat
     4801 acttaaagaa cggtatcatc tcttcaggtg ttcatgttgt cttggcccac atgtttttct
     4861 tgttaggtga cggtattaca caagaatcag ttgatttggt agatgactat ccaggtattt
     4921 ccacaagtat cgcaaccatt ttgagattat ctgatgactt gggttcagcc aaagatgaag
     4981 accaagatgg ttatgatggt tcttacatcg aatgttacat gaaggaacat aagggttcca
     5041 gtgtcgattc agccagagaa gaagtaataa gaatgatctc cgaagcatgg aaatgtttga
     5101 ataaggaatg cttatcacca aacccttttt ctgaatcatt cagaataggt tccttgaata
     5161 tggctagaat gatccctatg atgtactctt acgatgacaa ccataacttg ccaattttag
     5221 aagaacacat gaaggcaatg atctataaca catctttata actcgagata gattaattta
     5281 aacagtatat gtacagtttt atatatatat atatatatat acatatataa agaaacctgt
     5341 gcgttttttg tattttcaaa tacatttagt tttggcgcca cttctataaa aggtctcatg
     5401 atattgttac tctagagatc ccaatacaac agatcacgtg atctttttgt aagatgaagt
     5461 tgaagtagtg ttgcaccgtg ccaatgcagg tggctattag attaaatatg tgatttgttc
     5521 tattaagttt cctgtataat taatggggag cgctgattct cttttggtac gcttcccatc
     5581 cagcatttct gtatctttca ccttcaacct taggatctct acccttggcg aaaagtcctc
     5641 tgccaacaat gatgatatct gatccaccac ttacaacttc gtcgacggtt ctgtactgct
     5701 gacccaatgc atcgcctttg tcgtctaaac ctacacctgg ggtcatgatt agccaatcaa
     5761 acccttcttc tcttcctccc atatcgttct gagcaatgaa cccaataacg aaatctttat
     5821 cactctttgc aatatcaacg gtacccttag tatattcacc gtgtgctaga gaacccttgg
     5881 aagacaattc agcaagcatc aataatcccc ttggttcttt ggtgacctct tgcgcacctt
     5941 gtttcaagcc agcaacaata ccagcaccag taaccccgtg ggcgttggtg atatcagacc
     6001 attctgcgat acggtaaacg cccgatgtat attgtaattt gactgtgtta ccgatatcgg
     6061 cgaattttct gtcctcaaat atcaagaact tgtatttctc tgccaatgct ttcaatggaa
     6121 cgacagtacc ctcataactg aaatcatcca agatatcaac gtgtgttttc aaaaggcaaa
     6181 tgtatggacc caacgtttca acaagtttca atagctcatc agtcgaacga acgtcaagag
     6241 aagcacacaa attggtcttc ttttcatcca ttaaacgtaa aagtttcgat gcaaccggac
     6301 ttgcatgagt ctcagctcta ctggtatatg attttgtgga catgttttat cttcgtttcc
     6361 tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     6421 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     6481 gttcggagat taccgaatca aaaaaattta aatgaattca ctggccgtcg ttttacaacg
     6541 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6601 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6661 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6721 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6781 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6841 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6901 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6961 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8281 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     8461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8521 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8761 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8821 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     8881 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     8941 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     9001 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     9061 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     9121 aggaaacagc tatgaccatg attacgccaa gcttgcatgc atttaaatca ttacgaccga
     9181 gattcccggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     9241 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     9301 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     9361 gtcctactag ttatagtttt ttctccttga cgttaaagta tagaggt
//
