LOCUS       sequence-353105-        8310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing ERG20 (GAL10 promoter) and 
            nerolidol synthase from Actinidia chinensis (AcNES1; GAL7 promoter).
            Contains uracil auxotrophic marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  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 8310)
  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 NES
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(235..251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(235..251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(248..270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    259..275
                     /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        complement(join(283..289,290..307,308..313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    328..349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(466..483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(637..1225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(717..736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1396..2187,2188..2256))
                     /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     2019..2038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2257..2361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2429..2447
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2485..2507)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2607..2626
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2820..2842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2834..2851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2835..2851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7857..8298
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7857..7974
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     8269..8292
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
       61 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
      121 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
      181 cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat catggtcata
      241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     2461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2821 ccagtcacga cgttgtaaaa cgacggccag tgaattcatt taaatttttt tgattcggta
     2881 atctccgagc agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2941 atatgtggtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     3001 aaaacctgca ggaaacgaag ataaaacatg tccacaaaat catataccag tagagctgag
     3061 actcatgcaa gtccggttgc atcgaaactt ttacgtttaa tggatgaaaa gaagaccaat
     3121 ttgtgtgctt ctcttgacgt tcgttcgact gatgagctat tgaaacttgt tgaaacgttg
     3181 ggtccataca tttgcctttt gaaaacacac gttgatatct tggatgattt cagttatgag
     3241 ggtactgtcg ttccattgaa agcattggca gagaaataca agttcttgat atttgaggac
     3301 agaaaattcg ccgatatcgg taacacagtc aaattacaat atacatcggg cgtttaccgt
     3361 atcgcagaat ggtctgatat caccaacgcc cacggggtta ctggtgctgg tattgttgct
     3421 ggcttgaaac aaggtgcgca agaggtcacc aaagaaccaa ggggattatt gatgcttgct
     3481 gaattgtctt ccaagggttc tctagcacac ggtgaatata ctaagggtac cgttgatatt
     3541 gcaaagagtg ataaagattt cgttattggg ttcattgctc agaacgatat gggaggaaga
     3601 gaagaagggt ttgattggct aatcatgacc ccaggtgtag gtttagacga caaaggcgat
     3661 gcattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc agatatcatc
     3721 attgttggca gaggactttt cgccaagggt agagatccta aggttgaagg tgaaagatac
     3781 agaaatgctg gatgggaagc gtaccaaaag agaatcagcg ctccccatta attatacagg
     3841 aaacttaata gaacaaatca catatttaat ctaatagcca cctgcattgg cacggtgcaa
     3901 cactcacttc aacttcatct tacaaaagat cacgtgatct gttgtattgg gatctctaga
     3961 gtaacaatat catgagacct tttatagaag tggcgccaaa actaaatgta tttgaaaata
     4021 caaaaaacgc acaggtttct ttatatatgt atatatatat atatatatat ataaaactgt
     4081 acatatactg tttaaattaa tctatctcga gttataaaga tgtgttatag atcattgcct
     4141 tcatgtgttc ttctaaaatt ggcaagttat ggttgtcatc gtaagagtac atcataggga
     4201 tcattctagc catattcaag gaacctattc tgaatgattc agaaaaaggg tttggtgata
     4261 agcattcctt attcaaacat ttccatgctt cggagatcat tcttattact tcttctctgg
     4321 ctgaatcgac actggaaccc ttatgttcct tcatgtaaca ttcgatgtaa gaaccatcat
     4381 aaccatcttg gtcttcatct ttggctgaac ccaagtcatc agataatctc aaaatggttg
     4441 cgatacttgt ggaaatacct ggatagtcat ctaccaaatc aactgattct tgtgtaatac
     4501 cgtcacctaa caagaaaaac atgtgggcca agacaacatg aacacctgaa gagatgatac
     4561 cgttctttaa gtattcttct gcatttggca agtgaccaga ggcgaaccac tttgcttcaa
     4621 ctaaaaatgc attgcacaaa ctagcccaag tcttttgcaa ggaatcaata gggttttgac
     4681 catgcttttt atagattttg tatgcgattt cgttggtaac atcgtacaaa gccttgaaac
     4741 atactttcat gtagtctggt aattgttcga cggcagacaa ttcccatcta ttaactgcat
     4801 cagtaaacaa ggtcaattct tcaatggtac catagacgtc gaagatgtca tctatgatgt
     4861 atataaaaga aataggctta gttaattcaa ctctttcttc actgaatctt ggatcggtta
     4921 aagcggccat aggccacata taccacttca atggttggtt tctagccaat ttcaattcat
     4981 tggctaaacc tgtatccttc caccattgag aaacttggaa tacttccttt tgatggatac
     5041 tttgtacgat agtcaaatcc atcttagcta attcttgcaa attatcgaca ccgtcccagt
     5101 ttggaccctt gcagttcatg taggacaatt tttgacccat aaaggtagcc aatgacttgt
     5161 ggtatggatg tgttaaagtg ttagaaacca atctggcttc gtgatgatta ggatcttttg
     5221 tccatctacc taacaattga ctggagaaat cagcagcttg atctaagatg tcttcacctt
     5281 ctattctcaa ttgacttgct tcgtataagg acatcaaacc tctaatatct gttgacaatt
     5341 ctgatctgaa tctaccttcc ttatccttga agttattaaa gacgtcagcg gtaacattgt
     5401 gaccttcttg tcttaagaat ctaaacaatg ttgaaacttc gtacaaacca tgttcagggt
     5461 aaccatctgg agttgaagat atgatttgtt gcttttgcaa gaatgcacca atttcacctc
     5521 tgaaatggta atcgataccc aatctttgca aagcgtctat cataactaaa ccttccaaag
     5581 ggttttcttc aacttctctt aatgcttttc taacatcttc caacttttct tcgtacttaa
     5641 cgtataattc gtcagaaaat gatctgtaac cagtttgagg gttattgtgc ttactaggat
     5701 tggaaataca tgtcaaatct tcagctatag accatttatg agtaattggt acggaaggag
     5761 cgattggagt tctgtaagcg ttggagtttt gtggggagtt gttagttgcg ataggacctg
     5821 ctgcggtagc cattttgagc tcttttgagg gaatattcaa ctgttttttt ttatcatgtt
     5881 gatgctctgc ataataatgc ccataaatat ttccgacctg cttttatatc tttgctagcc
     5941 aaactaactg aacatagcta cacattattt tcagcttggc tattttgtga acactgtata
     6001 gccagtcctt cggatcacgg tcaacagttg tccgagcgct ttttggaccc tttcccttat
     6061 ttttgggtta aggaaaatga cagaaaatat atctaatgag ccttcgctca acagtgctcc
     6121 gaagtatagc tttccaaaag gagaggcaaa gcaatttaag aatgtatgaa caaaataaag
     6181 gggaaaaatt accccctcta ctttaccaaa cgaatactac caataatatt tacaactttt
     6241 ccttatgatt ttttcactga agcgcttcgc aatagttgtg agtgatatca aaagtaacga
     6301 aatgaactcc gcggctcgtg ctatattctt gttgctaccg tccatatctt tccatagatt
     6361 ttcaattttt gatgtctcca tggtggtaca gagaacttgt aaacaattcg gtccctacat
     6421 gtgaggaaat tcgctgtgac acttttatca ctgaactcca aatttaaaaa atagcataaa
     6481 attcgttata cagcaaatct atgtgttgca attaagaact aaaagatata gagtgcatat
     6541 tttcaagaag gatagtaagc tggcaaaaga tctttatttg cttctcttgt aaactttgtt
     6601 caaaaacgca gttaagacgt cggctttgaa gccacgagac tcgtcgactt gggagatctt
     6661 ggccttcaaa tccttggcaa cagactcttc atattcgtgg tataactggt cgattttcaa
     6721 gtcattgaaa atctttttgc atttggcttc tgcgactgag tccttcttac cgtaattttc
     6781 gtctaaagtc tttctttgtt ctgcggaagc aagttctaat gccttgttga ttacccaaga
     6841 acatttgtta tcttggatat ctgtaccgat cttaccgatc tgttctgggg taccgaagca
     6901 gtctaagtag tcatcttgaa tttggaaata ttcacccaat ggaatcaaga catctctggc
     6961 ttgtttcaaa tccttttcat ctgtgatacc ggcaacgtac atagccaatg cgacaggcaa
     7021 gtagaaagaa tagtaagcag tcttgaaagt aactatgaag gagtgcttct ttagggagaa
     7081 cttactcaag tcgactttgt cttcaggtgc agtgattaag tccatcaatt ggcccaattc
     7141 ggtttggaag gtgacttcat ggaacaattc ggtgatatct atgtagtatt tttcgtttct
     7201 gaagtgagat ttcaaaagct tgtagatagc agcctctaac atgaatgcgt cattgatggc
     7261 aatttcccca acttcaggaa ccttgtacca acatggttgg cctcttctgg taatggactt
     7321 gtccatcata tcatcggcga ccaagaagta agcctgcaac aactcaatgc accaacctag
     7381 aatagcaacc ttttcgtatt cttcttgccc caattgttca acggtcttgt tggagagaat
     7441 agcatacgtg tccacaacgg acaaacctct atttaactta ccgcctggag tgttgtagtt
     7501 caatgagtgg gcataccagt cacatgcttc cttaggcata ccgtaagcca aaagcgatgc
     7561 gttcaattcc tctactaatt tagggaaaac gttcaagaat ctctctctcc taatttcttt
     7621 ttctgaagcc attttggatc cttatattga attttcaaaa attcttactt tttttttgga
     7681 tggacgcaaa gaagtttaat aatcatatta catggcatta ccaccatata catatccata
     7741 tctaatctta cttatatgtt gtggaaatgt aaagagcccc attatcttag cctaaaaaaa
     7801 ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt gaagtacgga
     7861 ttagaagccg ccgagcgggc gacagccctc cgacggaaga ctctcctccg tgcgtcctcg
     7921 tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat
     7981 aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc
     8041 cccacaaacc ttcaaattaa cgaatcaaat taacaaccat aggatgataa tgcgattagt
     8101 tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca
     8161 gatatataaa tggaaaagct gcataaccac tttaactaat actttcaaca ttttcagttt
     8221 gtattacttc ttattcaaat gtcataaaag tatcaacaaa aaattgttaa tatacctcta
     8281 tactttaacg tcaaggagaa aaaactataa
//
