LOCUS       sequence-360431-        7554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20(F96W-N127W)-ClLIS1, with linker sequence AG4TGGA (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 7554)
  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 7554)
  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 ClLIS1 (M)
FEATURES             Location/Qualifiers
     source          1..7554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(73..514)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(79..102)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    397..514
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4764..4781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4764..4780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4773..4795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4989..5008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5108..5130
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5168..5186)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5254..5358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5359..5427,5428..6219)
                     /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(5577..5596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6390..6978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6879..6898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7132..7149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7266..7287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7302..7307,7308..7325,7326..7332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7340..7356
                     /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     7345..7367
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7364..7380
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7364..7380
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctactag
       61 ttatagtttt ttctccttga cgttaaagta tagaggtata ttaacaattt tttgttgata
      121 cttttatgac atttgaataa gaagtaatac aaaccgaaaa tgttgaaagt attagttaaa
      181 gtggttatgc agcttttgca tttatatatc tgttaataga tcaaaaatca tcgcttcgct
      241 gattaattac cccagaaata aggctaaaaa actaatcgca ttattatcct atggttgtta
      301 atttgattcg ttgatttgaa ggtttgtggg gccaggttac tgccaatttt tcctcttcat
      361 aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc gcgaggcaca
      421 tctgcgtttc aggaacgcga ccggtgaaga ccaggacgca cggaggagag tcttccgtcg
      481 gagggctgtc gcccgctcgg cggcttctaa tccgtacttc aatatagcaa tgagcagtta
      541 agcgtattac tgaaagttcc aaagagaagg tttttttagg ctaagataat ggggcttttt
      601 acatttccac aacatataag taagattaga tatggatatg tatatggtgg tattgccatg
      661 taatatgatt attaaacttc tttgcgtcca tccaaaaaaa aaagtaagaa tttttgaaaa
      721 ttcaatataa ggatccaaaa tggcttcaga aaaagaaatt aggagagaga gattcttgaa
      781 cgttttccct aaattagtag aggaattgaa cgcatcgctt ttggcttacg gtatgcctaa
      841 ggaagcatgt gactggtatg cccactcatt gaactacaac actccaggcg gtaagctaaa
      901 tagaggtttg tccgttgtgg acacgtatgc tattctctcc aacaagaccg ttgaacaatt
      961 ggggcaagaa gaatacgaaa aggttgccat tctaggttgg tgcattgagt tgttgcaggc
     1021 ttactggttg gtcgccgatg atatgatgga caagtccatt accagaagag gccaaccatg
     1081 ttggtacaag gttcctgaag ttggggaaat tgccatctgg gacgcattca tgttagaggc
     1141 tgctatctac aagcttttga aatctcactt cagaaacgaa aaatactaca tagatatcac
     1201 cgaattgttc catgaggtca ccttccaaac cgaattgggc caattgatgg acttaatcac
     1261 tgcacctgaa gacaaagtcg acttgagtaa gttctcccta aagaagcact ccttcatagt
     1321 tactttcaag actgcttact attctttcta cttgcctgtc gcattggcca tgtacgttgc
     1381 cggtatcacg gatgaaaagg atttgaaaca agccagagat gtcttgattc cattgggtga
     1441 atacttccaa attcaagatg actacttaga ctgcttcggt accccagaac agatcggtaa
     1501 gatcggtaca gatatccaag ataacaaatg ttcttgggta atcaacaagg cattagaact
     1561 tgcttccgca gaacaaagaa agactttaga cgaaaattac ggtaagaagg actcagtcgc
     1621 agaagccaaa tgcaaaaaga ttttcaatga cttgaaaatt gaacagctat accacgaata
     1681 tgaagagtct attgccaagg atttgaaggc caaaatttct caggtcgatg agtctcgtgg
     1741 cttcaaagct gatgtcttaa ctgcgttctt gaacaaagtt tacaagagaa gcaaaagatc
     1801 tgctggtgga ggaggtactg gtggtgctga gctcatgaga agatcagcta actatcaacc
     1861 atccatttgg gaccacgact ttttacaatc cttgaactct aactacaccg acgaagcata
     1921 caagagaaga gcagaagaat tacgtggtaa agtaaagata gccatcaaag atgtcatcga
     1981 acctttggac caattggaat tgattgataa cttgcaaaga ttgggtttag cccatagatt
     2041 tgaaaccgaa atcagaaaca tcttgaacaa catctataac aacaataagg attacaactg
     2101 gagaaaggaa aatttgtacg ctacttcctt ggaattcaga ttgttaagac aacacggtta
     2161 cccagtcagt caagaagttt ttaacggttt caaggatgac caaggtggtt ttatttgtga
     2221 tgacttcaag ggtatattgt ccttgcatga agcttcttac tactcattgg aaggtgaaag
     2281 tataatggaa gaagcatggc aattcacttc caaacacttg aaggaagtta tgataagtaa
     2341 aaatatggaa gaagatgttt tcgtagctga acaagcaaag agagccttgg aattaccatt
     2401 gcattggaaa gttcctatgt tggaagcaag atggttcatc catatatacg aaagaagaga
     2461 agataagaac cacttgttgt tggaattggc aaagatggaa ttcaatacat tacaagccat
     2521 ctatcaagaa gaattgaagg aaatctctgg ttggtggaag gataccggtt tgggtgaaaa
     2581 gttgtcattc gctagaaata gattggtcgc atctttctta tggtcaatgg gtattgcctt
     2641 tgaacctcaa ttcgcttact gtagaagagt tttgacaatc tctattgcat tgatcaccgt
     2701 tatagatgac atctatgacg tatacggtac tttggatgaa ttggaaatct ttacagacgc
     2761 tgttgaaaga tgggatatca actatgcatt aaagcatttg ccaggttaca tgaagatgtg
     2821 ctttttagcc ttgtacaact tcgtcaacga attcgcttac tacgttttga agcaacaaga
     2881 tttcgactta ttgttatcta ttaaaaacgc ttggttgggt ttgatacaag cctatttggt
     2941 agaggctaag tggtaccatt ctaagtacac acctaagttg gaagaatact tggaaaacgg
     3001 tttagtctca atcactggtc cattgatcat cacaatctcc tatttgagtg gtactaaccc
     3061 tatcattaaa aaggaattgg aattcttgga atcaaaccca gatatcgttc actggtcttc
     3121 aaaaattttc agattgcaag atgacttagg tacttccagt gacgaaattc aaagaggtga
     3181 cgtacctaaa tctatacaat gctacatgca tgaaacaggt gcatcagaag aagttgccag
     3241 acaacacatt aaggatatga tgagacaaat gtggaaaaag gttaatgctt acaccgcaga
     3301 taaagactcc ccattgactg gtactacaac cgaattcttg ttgaacttag ttagaatgtc
     3361 tcatttcatg tatttgcatg gtgacggtca cggtgtacaa aatcaagaaa ccattgatgt
     3421 cggttttact ttgttattcc aacctattcc attagaagat aaacacatgg cattcaccgc
     3481 atcaccaggt acaaagggtt aatagctcga gatagattaa tttaaacagt atatgtacag
     3541 ttttatatat atatatatat atatacatat ataaagaaac ctgtgcgttt tttgtatttt
     3601 caaatacatt tagttttggc gccacttcta taaaaggtct catgatattg ttactctaga
     3661 gatcccaata caacagatca cgtgatcttt ttgtaagatg aagttgaagt agtgttgcac
     3721 cgtgccaatg caggtggcta ttagattaaa tatgtgattt gttctattaa gtttcctgta
     3781 taattaatgg ggagcgctga ttctcttttg gtacgcttcc catccagcat ttctgtatct
     3841 ttcaccttca accttaggat ctctaccctt ggcgaaaagt cctctgccaa caatgatgat
     3901 atctgatcca ccacttacaa cttcgtcgac ggttctgtac tgctgaccca atgcatcgcc
     3961 tttgtcgtct aaacctacac ctggggtcat gattagccaa tcaaaccctt cttctcttcc
     4021 tcccatatcg ttctgagcaa tgaacccaat aacgaaatct ttatcactct ttgcaatatc
     4081 aacggtaccc ttagtatatt caccgtgtgc tagagaaccc ttggaagaca attcagcaag
     4141 catcaataat ccccttggtt ctttggtgac ctcttgcgca ccttgtttca agccagcaac
     4201 aataccagca ccagtaaccc cgtgggcgtt ggtgatatca gaccattctg cgatacggta
     4261 aacgcccgat gtatattgta atttgactgt gttaccgata tcggcgaatt ttctgtcctc
     4321 aaatatcaag aacttgtatt tctctgccaa tgctttcaat ggaacgacag taccctcata
     4381 actgaaatca tccaagatat caacgtgtgt tttcaaaagg caaatgtatg gacccaacgt
     4441 ttcaacaagt ttcaatagct catcagtcga acgaacgtca agagaagcac acaaattggt
     4501 cttcttttca tccattaaac gtaaaagttt cgatgcaacc ggacttgcat gagtctcagc
     4561 tctactggta tatgattttg tggacatgtt ttatcttcgt ttcctgcagg tttttgttct
     4621 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     4681 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     4741 atcaaaaaaa tttaaatgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4801 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4861 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4921 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4981 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5041 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5101 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5161 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5221 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5341 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5401 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5461 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5521 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5581 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5641 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5701 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5761 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5821 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5881 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5941 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6001 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6061 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7081 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7141 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7201 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7261 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7321 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7381 catgattacg ccaagcttgc atgcatttaa atcattacga ccgagattcc cgggtaataa
     7441 ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa
     7501 tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgc
//
