LOCUS       sequence-360448-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20-AcNES1, with linker sequence PTPTPTPTP (GAL10 promoter). 
            Contains uracil auxotrophic marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  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 7605)
  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: Linked Free (PT)4P
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(105..546)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(111..134)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    429..546
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4847..4864)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4847..4863)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4856..4878)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5072..5091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5191..5213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5251..5269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5337..5441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5442..5510,5511..6302)
                     /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(5660..5679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6473..7061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6962..6981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7215..7232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7349..7370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7385..7390,7391..7408,7409..7415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7423..7439
                     /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     7428..7450
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7447..7463
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7447..7463
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
       61 gcatcccttc cctttgcaaa tagtcctact agttatagtt ttttctcctt gacgttaaag
      121 tatagaggta tattaacaat tttttgttga tacttttatg acatttgaat aagaagtaat
      181 acaaaccgaa aatgttgaaa gtattagtta aagtggttat gcagcttttg catttatata
      241 tctgttaata gatcaaaaat catcgcttcg ctgattaatt accccagaaa taaggctaaa
      301 aaactaatcg cattattatc ctatggttgt taatttgatt cgttgatttg aaggtttgtg
      361 gggccaggtt actgccaatt tttcctcttc ataaccataa aagctagtat tgtagaatct
      421 ttattgttcg gagcagtgcg gcgcgaggca catctgcgtt tcaggaacgc gaccggtgaa
      481 gaccaggacg cacggaggag agtcttccgt cggagggctg tcgcccgctc ggcggcttct
      541 aatccgtact tcaatatagc aatgagcagt taagcgtatt actgaaagtt ccaaagagaa
      601 ggttttttta ggctaagata atggggctct ttacatttcc acaacatata agtaagatta
      661 gatatggata tgtatatggt ggtattgcca tgtaatatga ttattaaact tctttgcgtc
      721 catccaaaaa aaaaagtaag aatttttgaa aattcaatat aaggatccaa aatggcttca
      781 gaaaaagaaa ttaggagaga gagattcttg aacgttttcc ctaaattagt agaggaattg
      841 aacgcatcgc ttttggctta cggtatgcct aaggaagcat gtgactggta tgcccactca
      901 ttgaactaca acactccagg cggtaagtta aatagaggtt tgtccgttgt ggacacgtat
      961 gctattctct ccaacaagac cgttgaacaa ttggggcaag aagaatacga aaaggttgct
     1021 attctaggtt ggtgcattga gttgttgcag gcttacttct tggtcgccga tgatatgatg
     1081 gacaagtcca ttaccagaag aggccaacca tgttggtaca aggttcctga agttggggaa
     1141 attgccatca atgacgcatt catgttagag gctgctatct acaagctttt gaaatctcac
     1201 ttcagaaacg aaaaatacta catagatatc accgaattgt tccatgaagt caccttccaa
     1261 accgaattgg gccaattgat ggacttaatc actgcacctg aagacaaagt cgacttgagt
     1321 aagttctccc taaagaagca ctccttcata gttactttca agactgctta ctattctttc
     1381 tacttgcctg tcgcattggc tatgtacgtt gccggtatca cagatgaaaa ggatttgaaa
     1441 caagccagag atgtcttgat tccattgggt gaatatttcc aaattcaaga tgactactta
     1501 gactgcttcg gtaccccaga acagatcggt aagatcggta cagatatcca agataacaaa
     1561 tgttcttggg taatcaacaa ggcattagaa cttgcttccg cagaacaaag aaagacttta
     1621 gacgaaaatt acggtaagaa ggactcagtc gcagaagcca aatgcaaaaa gattttcaat
     1681 gacttgaaaa tcgaccagtt ataccacgaa tatgaagagt ctgttgccaa ggatttgaag
     1741 gccaagatct cccaagtcga cgagtctcgt ggcttcaaag ccgacgtctt aactgcgttt
     1801 ttgaacaaag tttacaagag aagcaaaaga tctccaacac ctactcccac accaacgcct
     1861 gagctcatgg ctaccgcagc aggtcctatc gcaactaaca actccccaca aaactccaac
     1921 gcttacagaa ctccaatcgc tccttccgta ccaattactc ataaatggtc tatagctgaa
     1981 gatttgacat gtatttccaa tcctagtaag cacaataacc ctcaaactgg ttacagatca
     2041 ttttctgacg aattatacgt taagtacgaa gaaaagttgg aagatgttag aaaagcatta
     2101 agagaagttg aagaaaaccc tttggaaggt ttagttatga tagacgcttt gcaaagattg
     2161 ggtatcgatt accatttcag aggtgaaatt ggtgcattct tgcaaaagca acaaatcata
     2221 tcttcaactc cagatggtta ccctgaacat ggtttgtacg aagtttcaac attgtttaga
     2281 ttcttaagac aagaaggtca caatgttacc gctgacgtct ttaataactt caaggataag
     2341 gaaggtagat tcagatcaga attgtcaaca gatattagag gtttgatgtc cttatacgaa
     2401 gcaagtcaat tgagaataga aggtgaagac atcttagatc aagctgctga tttctccagt
     2461 caattgttag gtagatggac aaaagatcct aatcatcacg aagccagatt ggtttctaac
     2521 actttaacac atccatacca caagtcattg gctaccttta tgggtcaaaa attgtcctac
     2581 atgaactgca agggtccaaa ctgggacggt gtcgataatt tgcaagaatt agctaagatg
     2641 gatttgacta tcgtacaaag tatccatcaa aaggaagtat tccaagtttc tcaatggtgg
     2701 aaggatacag gtttagccaa tgaattgaaa ttggctagaa accaaccatt gaagtggtat
     2761 atgtggccta tggccgcttt aaccgatcca agattcagtg aagaaagagt tgaattaact
     2821 aagcctattt cttttatata catcatagat gacatcttcg acgtctatgg taccattgaa
     2881 gaattgacct tgtttactga tgcagttaat agatgggaat tgtctgccgt cgaacaatta
     2941 ccagactaca tgaaagtatg tttcaaggct ttgtacgatg ttaccaacga aatcgcatac
     3001 aaaatctata aaaagcatgg tcaaaaccct attgattcct tgcaaaagac ttgggctagt
     3061 ttgtgcaatg catttttagt tgaagcaaag tggttcgcct ctggtcactt gccaaatgca
     3121 gaagaatact taaagaacgg tatcatctct tcaggtgttc atgttgtctt ggcccacatg
     3181 tttttcttgt taggtgacgg tattacacaa gaatcagttg atttggtaga tgactatcca
     3241 ggtatttcca caagtatcgc aaccattttg agattatctg atgacttggg ttcagccaaa
     3301 gatgaagacc aagatggtta tgatggttct tacatcgaat gttacatgaa ggaacataag
     3361 ggttccagtg tcgattcagc cagagaagaa gtaataagaa tgatctccga agcatggaaa
     3421 tgtttgaata aggaatgctt atcaccaaac cctttttctg aatcattcag aataggttcc
     3481 ttgaatatgg ctagaatgat ccctatgatg tactcttacg atgacaacca taacttgcca
     3541 attttagaag aacacatgaa ggcaatgatc tataacacat ctttataact cgagatagat
     3601 taatttaaac agtatatgta cagttttata tatatatata tatatataca tatataaaga
     3661 aacctgtgcg ttttttgtat tttcaaatac atttagtttt ggcgccactt ctataaaagg
     3721 tctcatgata ttgttactct agagatccca atacaacaga tcacgtgatc tttttgtaag
     3781 atgaagttga agtagtgttg caccgtgcca atgcaggtgg ctattagatt aaatatgtga
     3841 tttgttctat taagtttcct gtataattaa tggggagcgc tgattctctt ttggtacgct
     3901 tcccatccag catttctgta tctttcacct tcaaccttag gatctctacc cttggcgaaa
     3961 agtcctctgc caacaatgat gatatctgat ccaccactta caacttcgtc gacggttctg
     4021 tactgctgac ccaatgcatc gcctttgtcg tctaaaccta cacctggggt catgattagc
     4081 caatcaaacc cttcttctct tcctcccata tcgttctgag caatgaaccc aataacgaaa
     4141 tctttatcac tctttgcaat atcaacggta cccttagtat attcaccgtg tgctagagaa
     4201 cccttggaag acaattcagc aagcatcaat aatccccttg gttctttggt gacctcttgc
     4261 gcaccttgtt tcaagccagc aacaatacca gcaccagtaa ccccgtgggc gttggtgata
     4321 tcagaccatt ctgcgatacg gtaaacgccc gatgtatatt gtaatttgac tgtgttaccg
     4381 atatcggcga attttctgtc ctcaaatatc aagaacttgt atttctctgc caatgctttc
     4441 aatggaacga cagtaccctc ataactgaaa tcatccaaga tatcaacgtg tgttttcaaa
     4501 aggcaaatgt atggacccaa cgtttcaaca agtttcaata gctcatcagt cgaacgaacg
     4561 tcaagagaag cacacaaatt ggtcttcttt tcatccatta aacgtaaaag tttcgatgca
     4621 accggacttg catgagtctc agctctactg gtatatgatt ttgtggacat gttttatctt
     4681 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     4741 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     4801 tccttctgtt cggagattac cgaatcaaaa aaatttaaat gaattcactg gccgtcgttt
     4861 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4921 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4981 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     5041 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5101 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5161 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5221 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5281 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5341 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5401 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5461 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5521 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5581 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5641 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5701 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5761 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5821 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5881 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5941 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6001 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6061 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6121 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6181 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6241 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6301 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6361 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6601 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7141 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7201 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7261 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7321 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7381 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7441 ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcatt taaatcatta
     7501 cgaccgagat tcccgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     7561 agtatacatg catttactta taatacagtt ttttagtttt gctgg
//
