LOCUS       sequence-360434-        7632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20-AcNES1, with linker sequence (AG4TGGA)2 (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 7632)
  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 7632)
  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 NES (AG4TGGA)2
FEATURES             Location/Qualifiers
     source          1..7632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(24..465)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(30..53)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    348..465
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4793..4810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4793..4809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4802..4824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5018..5037)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5137..5159
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5197..5215)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5283..5387
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5388..5456,5457..6248)
                     /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(5606..5625)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6419..7007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6908..6927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7161..7178
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7295..7316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7331..7336,7337..7354,7355..7361)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7369..7385
                     /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     7374..7396
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7393..7409
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7393..7409
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agtcctacta gttatagttt tttctccttg acgttaaagt atagaggtat attaacaatt
       61 ttttgttgat acttttatga catttgaata agaagtaata caaaccgaaa atgttgaaag
      121 tattagttaa agtggttatg cagcttttgc atttatatat ctgttaatag atcaaaaatc
      181 atcgcttcgc tgattaatta ccccagaaat aaggctaaaa aactaatcgc attattatcc
      241 tatggttgtt aatttgattc gttgatttga aggtttgtgg ggccaggtta ctgccaattt
      301 ttcctcttca taaccataaa agctagtatt gtagaatctt tattgttcgg agcagtgcgg
      361 cgcgaggcac atctgcgttt caggaacgcg accggtgaag accaggacgc acggaggaga
      421 gtcttccgtc ggagggctgt cgcccgctcg gcggcttcta atccgtactt caatatagca
      481 atgagcagtt aagcgtatta ctgaaagttc caaagagaag gtttttttag gctaagataa
      541 tggggctctt tacatttcca caacatataa gtaagattag atatggatat gtatatggtg
      601 gtattgccat gtaatatgat tattaaactt ctttgcgtcc atccaaaaaa aaaagtaaga
      661 atttttgaaa attcaatata aggatccaaa atggcttcag aaaaagaaat taggagagag
      721 agattcttga acgttttccc taaattagta gaggaattga acgcatcgct tttggcttac
      781 ggtatgccta aggaagcatg tgactggtat gcccactcat tgaactacaa cactccaggc
      841 ggtaagttaa atagaggttt gtccgttgtg gacacgtatg ctattctctc caacaagacc
      901 gttgaacaat tggggcaaga agaatacgaa aaggttgcta ttctaggttg gtgcattgag
      961 ttgttgcagg cttacttctt ggtcgccgat gatatgatgg acaagtccat taccagaaga
     1021 ggccaaccat gttggtacaa ggttcctgaa gttggggaaa ttgccatcaa tgacgcattc
     1081 atgttagagg ctgctatcta caagcttttg aaatctcact tcagaaacga aaaatactac
     1141 atagatatca ccgaattgtt ccatgaagtc accttccaaa ccgaattggg ccaattgatg
     1201 gacttaatca ctgcacctga agacaaagtc gacttgagta agttctccct aaagaagcac
     1261 tccttcatag ttactttcaa gactgcttac tattctttct acttgcctgt cgcattggct
     1321 atgtacgttg ccggtatcac agatgaaaag gatttgaaac aagccagaga tgtcttgatt
     1381 ccattgggtg aatatttcca aattcaagat gactacttag actgcttcgg taccccagaa
     1441 cagatcggta agatcggtac agatatccaa gataacaaat gttcttgggt aatcaacaag
     1501 gcattagaac ttgcttccgc agaacaaaga aagactttag acgaaaatta cggtaagaag
     1561 gactcagtcg cagaagccaa atgcaaaaag attttcaatg acttgaaaat cgaccagtta
     1621 taccacgaat atgaagagtc tgttgccaag gatttgaagg ccaagatctc ccaagtcgac
     1681 gagtctcgtg gcttcaaagc cgacgtctta actgcgtttt tgaacaaagt ttacaagaga
     1741 agcaaaagat ctgctggtgg aggaggtact ggtggtgctg caggaggcgg aggtacagga
     1801 ggtgcagagc tcatggctac cgcagcaggt cctatcgcaa ctaacaactc cccacaaaac
     1861 tccaacgctt acagaactcc aatcgctcct tccgtaccaa ttactcataa atggtctata
     1921 gctgaagatt tgacatgtat ttccaatcct agtaagcaca ataaccctca aactggttac
     1981 agatcatttt ctgacgaatt atacgttaag tacgaagaaa agttggaaga tgttagaaaa
     2041 gcattaagag aagttgaaga aaaccctttg gaaggtttag ttatgataga cgctttgcaa
     2101 agattgggta tcgattacca tttcagaggt gaaattggtg cattcttgca aaagcaacaa
     2161 atcatatctt caactccaga tggttaccct gaacatggtt tgtacgaagt ttcaacattg
     2221 tttagattct taagacaaga aggtcacaat gttaccgctg acgtctttaa taacttcaag
     2281 gataaggaag gtagattcag atcagaattg tcaacagata ttagaggttt gatgtcctta
     2341 tacgaagcaa gtcaattgag aatagaaggt gaagacatct tagatcaagc tgctgatttc
     2401 tccagtcaat tgttaggtag atggacaaaa gatcctaatc atcacgaagc cagattggtt
     2461 tctaacactt taacacatcc ataccacaag tcattggcta cctttatggg tcaaaaattg
     2521 tcctacatga actgcaaggg tccaaactgg gacggtgtcg ataatttgca agaattagct
     2581 aagatggatt tgactatcgt acaaagtatc catcaaaagg aagtattcca agtttctcaa
     2641 tggtggaagg atacaggttt agccaatgaa ttgaaattgg ctagaaacca accattgaag
     2701 tggtatatgt ggcctatggc cgctttaacc gatccaagat tcagtgaaga aagagttgaa
     2761 ttaactaagc ctatttcttt tatatacatc atagatgaca tcttcgacgt ctatggtacc
     2821 attgaagaat tgaccttgtt tactgatgca gttaatagat gggaattgtc tgccgtcgaa
     2881 caattaccag actacatgaa agtatgtttc aaggctttgt acgatgttac caacgaaatc
     2941 gcatacaaaa tctataaaaa gcatggtcaa aaccctattg attccttgca aaagacttgg
     3001 gctagtttgt gcaatgcatt tttagttgaa gcaaagtggt tcgcctctgg tcacttgcca
     3061 aatgcagaag aatacttaaa gaacggtatc atctcttcag gtgttcatgt tgtcttggcc
     3121 cacatgtttt tcttgttagg tgacggtatt acacaagaat cagttgattt ggtagatgac
     3181 tatccaggta tttccacaag tatcgcaacc attttgagat tatctgatga cttgggttca
     3241 gccaaagatg aagaccaaga tggttatgat ggttcttaca tcgaatgtta catgaaggaa
     3301 cataagggtt ccagtgtcga ttcagccaga gaagaagtaa taagaatgat ctccgaagca
     3361 tggaaatgtt tgaataagga atgcttatca ccaaaccctt tttctgaatc attcagaata
     3421 ggttccttga atatggctag aatgatccct atgatgtact cttacgatga caaccataac
     3481 ttgccaattt tagaagaaca catgaaggca atgatctata acacatcttt ataactcgag
     3541 atagattaat ttaaacagta tatgtacagt tttatatata tatatatata tatacatata
     3601 taaagaaacc tgtgcgtttt ttgtattttc aaatacattt agttttggcg ccacttctat
     3661 aaaaggtctc atgatattgt tactctagag atcccaatac aacagatcac gtgatctttt
     3721 tgtaagatga agttgaagta gtgttgcacc gtgccaatgc aggtggctat tagattaaat
     3781 atgtgatttg ttctattaag tttcctgtat aattaatggg gagcgctgat tctcttttgg
     3841 tacgcttccc atccagcatt tctgtatctt tcaccttcaa ccttaggatc tctacccttg
     3901 gcgaaaagtc ctctgccaac aatgatgata tctgatccac cacttacaac ttcgtcgacg
     3961 gttctgtact gctgacccaa tgcatcgcct ttgtcgtcta aacctacacc tggggtcatg
     4021 attagccaat caaacccttc ttctcttcct cccatatcgt tctgagcaat gaacccaata
     4081 acgaaatctt tatcactctt tgcaatatca acggtaccct tagtatattc accgtgtgct
     4141 agagaaccct tggaagacaa ttcagcaagc atcaataatc cccttggttc tttggtgacc
     4201 tcttgcgcac cttgtttcaa gccagcaaca ataccagcac cagtaacccc gtgggcgttg
     4261 gtgatatcag accattctgc gatacggtaa acgcccgatg tatattgtaa tttgactgtg
     4321 ttaccgatat cggcgaattt tctgtcctca aatatcaaga acttgtattt ctctgccaat
     4381 gctttcaatg gaacgacagt accctcataa ctgaaatcat ccaagatatc aacgtgtgtt
     4441 ttcaaaaggc aaatgtatgg acccaacgtt tcaacaagtt tcaatagctc atcagtcgaa
     4501 cgaacgtcaa gagaagcaca caaattggtc ttcttttcat ccattaaacg taaaagtttc
     4561 gatgcaaccg gacttgcatg agtctcagct ctactggtat atgattttgt ggacatgttt
     4621 tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     4681 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     4741 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttaaatgaat tcactggccg
     4801 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4861 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4921 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     4981 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     5041 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5101 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5161 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5221 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5281 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5341 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5401 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5461 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5521 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5581 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5641 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5701 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5761 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5821 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5881 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5941 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6001 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6061 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6121 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6181 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6241 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6301 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6361 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6421 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6481 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6541 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6601 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6661 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6721 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6781 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6841 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6901 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6961 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7021 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7081 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7141 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7201 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7261 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7321 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7381 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcatttaaa
     7441 tcattacgac cgagattccc gggtaataac tgatataatt aaattgaagc tctaatttgt
     7501 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     7561 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     7621 ccctttgcaa at
//
