LOCUS       sequence-360445-        8309 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 8309)
  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 8309)
  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..8309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..70
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(323..339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(323..339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(336..358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    347..363
                     /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(371..377,378..395,396..401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    416..437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(554..571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(725..1313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(805..824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1484..2275,2276..2344))
                     /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     2107..2126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2345..2449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2517..2535
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2573..2595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2695..2714
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2908..2930
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2922..2939
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2923..2939
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(7944..8309,1..76)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7944..8061
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
ORIGIN
        1 attacttctt attcaaatgt cataaaagta tcaacaaaaa attgttaata tacctctata
       61 ctttaacgtc aaggagaaaa aactataact agtaggacta tttgcaaagg gaagggatgc
      121 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
      181 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
      241 agcttcaatt taattatatc agttattacc cgggaatctc ggtcgtaatg atttaaatgc
      301 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      361 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      421 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      481 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2341 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2401 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2461 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2521 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2581 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2641 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2701 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2761 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     2821 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     2881 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2941 aattcattta aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     3001 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     3061 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaaacatgtc
     3121 cacaaaatca tataccagta gagctgagac tcatgcaagt ccggttgcat cgaaactttt
     3181 acgtttaatg gatgaaaaga agaccaattt gtgtgcttct cttgacgttc gttcgactga
     3241 tgagctattg aaacttgttg aaacgttggg tccatacatt tgccttttga aaacacacgt
     3301 tgatatcttg gatgatttca gttatgaggg tactgtcgtt ccattgaaag cattggcaga
     3361 gaaatacaag ttcttgatat ttgaggacag aaaattcgcc gatatcggta acacagtcaa
     3421 attacaatat acatcgggcg tttaccgtat cgcagaatgg tctgatatca ccaacgccca
     3481 cggggttact ggtgctggta ttgttgctgg cttgaaacaa ggtgcgcaag aggtcaccaa
     3541 agaaccaagg ggattattga tgcttgctga attgtcttcc aagggttctc tagcacacgg
     3601 tgaatatact aagggtaccg ttgatattgc aaagagtgat aaagatttcg ttattgggtt
     3661 cattgctcag aacgatatgg gaggaagaga agaagggttt gattggctaa tcatgacccc
     3721 aggtgtaggt ttagacgaca aaggcgatgc attgggtcag cagtacagaa ccgtcgacga
     3781 agttgtaagt ggtggatcag atatcatcat tgttggcaga ggacttttcg ccaagggtag
     3841 agatcctaag gttgaaggtg aaagatacag aaatgctgga tgggaagcgt accaaaagag
     3901 aatcagcgct ccccattaat tatacaggaa acttaataga acaaatcaca tatttaatct
     3961 aatagccacc tgcattggca cggtgcaaca ctacttcaac ttcatcttac aaaaagatca
     4021 cgtgatctgt tgtattggga tctctagagt aacaatatca tgagaccttt tatagaagtg
     4081 gcgccaaaac taaatgtatt tgaaaataca aaaaacgcac aggtttcttt atatatgtat
     4141 atatatatat atatatataa aactgtacat atactgttta aattaatcta tctcgagtta
     4201 taaagatgtg ttatagatca ttgccttcat gtgttcttct aaaattggca agttatggtt
     4261 gtcatcgtaa gagtacatca tagggatcat tctagccata ttcaaggaac ctattctgaa
     4321 tgattcagaa aaagggtttg gtgataagca ttccttattc aaacatttcc atgcttcgga
     4381 gatcattctt attacttctt ctctggctga atcgacactg gaacccttat gttccttcat
     4441 gtaacattcg atgtaagaac catcataacc atcttggtct tcatctttgg ctgaacccaa
     4501 gtcatcagat aatctcaaaa tggttgcgat acttgtggaa atacctggat agtcatctac
     4561 caaatcaact gattcttgtg taataccgtc acctaacaag aaaaacatgt gggccaagac
     4621 aacatgaaca cctgaagaga tgataccgtt ctttaagtat tcttctgcat ttggcaagtg
     4681 accagaggcg aaccactttg cttcaactaa aaatgcattg cacaaactag cccaagtctt
     4741 ttgcaaggaa tcaatagggt tttgaccatg ctttttatag attttgtatg cgatttcgtt
     4801 ggtaacatcg tacaaagcct tgaaacatac tttcatgtag tctggtaatt gttcgacggc
     4861 agacaattcc catctattaa ctgcatcagt aaacaaggtc aattcttcaa tggtaccata
     4921 gacgtcgaag atgtcatcta tgatgtatat aaaagaaata ggcttagtta attcaactct
     4981 ttcttcactg aatcttggat cggttaaagc ggccataggc cacatatacc acttcaatgg
     5041 ttggtttcta gccaatttca attcattggc taaacctgta tccttccacc attgagaaac
     5101 ttggaatact tccttttgat ggatactttg tacgatagtc aaatccatct tagctaattc
     5161 ttgcaaatta tcgacaccgt cccagtttgg acccttgcag ttcatgtagg acaatttttg
     5221 acccataaag gtagccaatg acttgtggta tggatgtgtt aaagtgttag aaaccaatct
     5281 ggcttcgtga tgattaggat cttttgtcca tctacctaac aattgactgg agaaatcagc
     5341 agcttgatct aagatgtctt caccttctat tctcaattga cttgcttcgt ataaggacat
     5401 caaacctcta atatctgttg acaattctga tctgaatcta ccttccttat ccttgaagtt
     5461 attaaagacg tcagcggtaa cattgtgacc ttcttgtctt aagaatctaa acaatgttga
     5521 aacttcgtac aaaccatgtt cagggtaacc atctggagtt gaagatatga tttgttgctt
     5581 ttgcaagaat gcaccaattt cacctctgaa atggtaatcg atacccaatc tttgcaaagc
     5641 gtctatcata actaaacctt ccaaagggtt ttcttcaact tctcttaatg cttttctaac
     5701 atcttccaac ttttcttcgt acttaacgta taattcgtca gaaaatgatc tgtaaccagt
     5761 ttgagggtta ttgtgcttac taggattgga aatacatgtc aaatcttcag ctatagacca
     5821 tttatgagta attggtacgg aaggagcgat tggagttctg taagcgttgg agttttgtgg
     5881 ggagttgtta gttgcgatag gacctgctgc ggtagccatt ttgagctctt ttgagggaat
     5941 attcaactgt ttttttttat catgttgatg ctctgcataa taatgcccat aaatatttcc
     6001 gacctgcttt tatatctttg ctagccaaac taactgaaca tagctacaca ttattttcag
     6061 cttggctatt ttgtgaacac tgtatagcca gtccttcgga tcacggtcaa cagttgtccg
     6121 agcgcttttt ggaccctttc ccttattttt gggttaagga aaatgacaga aaatatatct
     6181 aatgagcctt cgctcaacag tgctccgaag tatagctttc caaaaggaga ggcaaagcaa
     6241 tttaagaatg tatgaacaaa ataaagggga aaaattaccc cctctacttt accaaacgaa
     6301 tactaccaat aatatttaca acttttcctt atgatttttt cactgaagcg cttcgcaata
     6361 gttgtgagtg atatcaaaag taacgaaatg aactccgcgg ctcgtgctat attcttgttg
     6421 ctaccgtcca tatctttcca tagattttca atttttgatg tctccatggt ggtacagaga
     6481 acttgtaaac aattcggtcc ctacatgtga ggaaattcgc tgtgacactt ttatcactga
     6541 actccaaatt taaaaaatag cataaaattc gttatacagc aaatctatgt gttgcaatta
     6601 agaactaaaa gatatagagt gcatattttc aagaaggata gtaagctggc aaaagatctt
     6661 tatttgcttc tcttgtaaac tttgttcaaa aacgcagtta agacgtcggc tttgaagcca
     6721 cgagactcgt cgacttggga gatcttggcc ttcaaatcct tggcaacaga ctcttcatat
     6781 tcgtggtata actggtcgat tttcaagtca ttgaaaatct ttttgcattt ggcttctgcg
     6841 actgagtcct tcttaccgta attttcgtct aaagtctttc tttgttctgc ggaagcaagt
     6901 tctaatgcct tgttgattac ccaagaacat ttgttatctt ggatatctgt accgatctta
     6961 ccgatctgtt ctggggtacc gaagcagtct aagtagtcat cttgaatttg gaaatattca
     7021 cccaatggaa tcaagacatc tctggcttgt ttcaaatcct tttcatctgt gataccggca
     7081 acgtacatag ccaatgcgac aggcaagtag aaagaatagt aagcagtctt gaaagtaact
     7141 atgaaggagt gcttctttag ggagaactta ctcaagtcga ctttgtcttc aggtgcagtg
     7201 attaagtcca tcaattggcc caattcggtt tggaaggtga cttcatggaa caattcggtg
     7261 atatctatgt agtatttttc gtttctgaag tgagatttca aaagcttgta gatagcagcc
     7321 tctaacatga atgcgtcatt gatggcaatt tccccaactt caggaacctt gtaccaacat
     7381 ggttggcctc ttctggtaat ggacttgtcc atcatatcat cggcgaccaa gaagtaagcc
     7441 tgcaacaact caatgcacca acctagaata gcaacctttt cgtattcttc ttgccccaat
     7501 tgttcaacgg tcttgttgga gagaatagca tacgtgtcca caacggacaa acctctattt
     7561 aacttaccgc ctggagtgtt gtagttcaat gagtgggcat accagtcaca tgcttcctta
     7621 ggcataccgt aagccaaaag cgatgcgttc aattcctcta ctaatttagg gaaaacgttc
     7681 aagaatctct ctctcctaat ttctttttct gaagccattt tggatcctta tattgaattt
     7741 tcaaaaattc ttactttttt ttttggatgg acgcaaagaa gtttaataat catattacat
     7801 ggcaatacca ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
     7861 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
     7921 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga
     7981 cggaagactc tcctccgtgc gtcctggtct tcaccggtcg cgttcctgaa acgcagatgt
     8041 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
     8101 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatcaacga atcaaattaa
     8161 caaccatagg ataataatgc gattagtttt ttagccttat ttctggggta attaatcagc
     8221 gaagcgatga tttttgatct attaacagat atataaatgc aaaagctgca taaccacttt
     8281 aactaatact ttcaacattt tcggtttgt
//
