LOCUS       sequence-360437-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein ERG20-PcPTS,
            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 7542)
  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 7542)
  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 PcPTS
FEATURES             Location/Qualifiers
     source          1..7542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(166..607)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(172..195)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    490..607
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4845..4862)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4845..4861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4854..4876)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5070..5089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5189..5211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5249..5267)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5335..5439
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5440..5508,5509..6300)
                     /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(5658..5677)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6471..7059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6960..6979
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7213..7230
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7347..7368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7383..7388,7389..7406,7407..7413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7421..7437
                     /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     7426..7448
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7445..7461
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7445..7461
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
       61 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
      121 agcatccctt ccctttgcaa atagtcctac tagttatagt tttttctcct tgacgttaaa
      181 gtatagaggt atattaacaa ttttttgttg atacttttat gacatttgaa taagaagtaa
      241 tacaaaccga aaatgttgaa agtattagtt aaagtggtta tgcagctttt gcatttatat
      301 atctgttaat agatcaaaaa tcatcgcttc gctgattaat taccccagaa ataaggctaa
      361 aaaactaatc gcattattat cctatggttg ttaatttgat tcgttgattt gaaggtttgt
      421 ggggccaggt tactgccaat ttttcctctt cataaccata aaagctagta ttgtagaatc
      481 tttattgttc ggagcagtgc ggcgcgaggc acatctgcgt ttcaggaacg cgaccggtga
      541 agaccaggac gcacggagga gagtcttccg tcggagggct gtcgcccgct cggcggcttc
      601 taatccgtac ttcaatatag caatgagcag ttaagcgtat tactgaaagt tccaaagaga
      661 aggttttttt aggctaagat aatggggctc tttacatttc cacaacatat aagtaagatt
      721 agatatggat atgtatatgg tggtattgcc atgtaatatg attattaaac ttctttgcgt
      781 ccatccaaaa aaaaaagtaa gaatttttga aaattcaata taaggatcca aaatggcttc
      841 agaaaaagaa attaggagag agagattctt gaacgttttc cctaaattag tagaggaatt
      901 gaacgcatcg cttttggctt acggtatgcc taaggaagca tgtgactggt atgcccactc
      961 attgaactac aacactccag gcggtaagtt aaatagaggt ttgtccgttg tggacacgta
     1021 tgctattctc tccaacaaga ccgttgaaca attggggcaa gaagaatacg aaaaggttgc
     1081 tattctaggt tggtgcattg agttgttgca ggcttacttc ttggtcgccg atgatatgat
     1141 ggacaagtcc attaccagaa gaggccaacc atgttggtac aaggttcctg aagttgggga
     1201 aattgccatc aatgacgcat tcatgttaga ggctgctatc tacaagcttt tgaaatctca
     1261 cttcagaaac gaaaaatact acatagatat caccgaattg ttccatgaag tcaccttcca
     1321 aaccgaattg ggccaattga tggacttaat cactgcacct gaagacaaag tcgacttgag
     1381 taagttctcc ctaaagaagc actccttcat agttactttc aagactgctt actattcttt
     1441 ctacttgcct gtcgcattgg ctatgtacgt tgccggtatc acagatgaaa aggatttgaa
     1501 acaagccaga gatgtcttga ttccattggg tgaatatttc caaattcaag atgactactt
     1561 agactgcttc ggtaccccag aacagatcgg taagatcggt acagatatcc aagataacaa
     1621 atgttcttgg gtaatcaaca aggcattaga acttgcttcc gcagaacaaa gaaagacttt
     1681 agacgaaaat tacggtaaga aggactcagt cgcagaagcc aaatgcaaaa agattttcaa
     1741 tgacttgaaa atcgaccagt tataccacga atatgaagag tctgttgcca aggatttgaa
     1801 ggccaagatc tcccaagtcg acgagtctcg tggcttcaaa gccgacgtct taactgcgtt
     1861 tttgaacaaa gtttacaaga gaagcaaaag atctgctggt ggaggaggta ctggtggtgc
     1921 tgagctcatg gaattgtatg ctcaatctgt tggtgttggt gctgcatcaa gacctctagc
     1981 caattttcat caatgtgtat ggggagacaa attcattgtt tacaacccac aatcatccca
     2041 ggctggagag agagaacagg ctgaggagct taaagtagag ctaaaaagag agcttaagga
     2101 agcatcagat aattatatga ggcaactgaa aatggtagat gcaatacaac gtttaggcat
     2161 tgactattta tttgtcgaag atgttgatga agctttgaag aatttgtttg aaatgtttga
     2221 tgctttctgc aagaataatc atgacatgca cgccactgct ctatcctttc gtcttctaag
     2281 acaacatggc tatagagttt catgtgaagt ttttgaaaag tttaaggatg gcaaagatgg
     2341 atttaaggtt ccaaatgagg atggagccgt tgcagtcctt gaattcttcg aagccactca
     2401 tttgagagtc catggagaag acgtccttga taatgctttt gtcttcacta ggaactactt
     2461 ggaatcagtc tatgcaactt tgaacgatcc aaccgcgaat caagtccaca acgcattgaa
     2521 tgagttctct tttaggagag gattgccacg tgtggaagca aggaagtaca tatcaatcta
     2581 cgagcaatac gcatctcatc acaaaggctt gttaaaactt gctaagctgg atttcaactt
     2641 ggtacaagct ttgcacagaa gggagctgag tgaagattct aggtggtgga agactttaca
     2701 agtgcccaca gagctatcat tcgttagaga taggttggtg gagtcctact tttgggcttc
     2761 gggatcttat ttcgaaccga attattcggt agctaggatg attttagcaa aaggattagc
     2821 tgtattatct cttatggacg atgtgtatga tgcatatggt acttttgagg aattacaagt
     2881 gttcacagat gcaatcgaaa ggtgggatgc ttcatgttta gataaacttc cagagtacat
     2941 gaaaatagta tacaaggccc ttttggatgt gtttgaggaa gttgacgagg aggtgatcaa
     3001 gctaggtgca ccatataggg tctactatgg aaaagaagcc atgaaatatg ccgctagagc
     3061 ttacatggaa gaggcccaat ggagggagca aaagcacaaa cccacaacta aggagtatat
     3121 gaagctggca acaaagacct gtggctatat aactctaata atattatcat ttcttggagt
     3181 ggaagagggc attgtgacta aagaagcctt cgattgggtt ttctccagac ctcctttcgt
     3241 cgaggctact ttaatcattg ctaggttgat caatgatatt acaggatgtg aatttgagaa
     3301 taaaagggag cacgttagaa ctgcagtcga atgctacatg gaagagcaca aagtgggaaa
     3361 gcaagaggtg gtctctgaat tttacaacca aatggagtca gcatggaagg acataaatga
     3421 gtgcttgcta agaccagctg aatttccaat ccctctactt aatcttattt taaattctgt
     3481 cagaacactt gaggttattt acaaagaggg cgattcctat acacacgtgg gtcctgcaat
     3541 gcaaaacatc atcaagcagt tgtacctaca ccctgttcca tattaactcg agatagatta
     3601 atttaaacag tatatgtaca gttttatata tatatatata tatatacata tataaagaaa
     3661 cctgtgcgtt ttttgtattt tcaaatacat ttagttttgg cgccacttct ataaaaggtc
     3721 tcatgatatt gttactctag agatcccaat acaacagatc acgtgatctt tttgtaagat
     3781 gaagttgaag tagtgttgca ccgtgccaat gcaggtggct attagattaa atatgtgatt
     3841 tgttctatta agtttcctgt ataattaatg gggagcgctg attctctttt ggtacgcttc
     3901 ccatccagca tttctgtatc tttcaccttc aaccttagga tctctaccct tggcgaaaag
     3961 tcctctgcca acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta
     4021 ctgctgaccc aatgcatcgc ctttgtcgtc taaacctaca cctggggtca tgattagcca
     4081 atcaaaccct tcttctcttc ctcccatatc gttctgagca atgaacccaa taacgaaatc
     4141 tttatcactc tttgcaatat caacggtacc cttagtatat tcaccgtgtg ctagagaacc
     4201 cttggaagac aattcagcaa gcatcaataa tccccttggt tctttggtga cctcttgcgc
     4261 accttgtttc aagccagcaa caataccagc accagtaacc ccgtgggcgt tggtgatatc
     4321 agaccattct gcgatacggt aaacgcccga tgtatattgt aatttgactg tgttaccgat
     4381 atcggcgaat tttctgtcct caaatatcaa gaacttgtat ttctctgcca atgctttcaa
     4441 tggaacgaca gtaccctcat aactgaaatc atccaagata tcaacgtgtg ttttcaaaag
     4501 gcaaatgtat ggacccaacg tttcaacaag tttcaatagc tcatcagtcg aacgaacgtc
     4561 aagagaagca cacaaattgg tcttcttttc atccattaaa cgtaaaagtt tcgatgcaac
     4621 cggacttgca tgagtctcag ctctactggt atatgatttt gtggacatgt tttatcttcg
     4681 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
     4741 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     4801 cttctgttcg gagattaccg aatcaaaaaa atttaaatga attcactggc cgtcgtttta
     4861 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4921 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4981 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5041 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5101 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5161 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5221 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5281 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5341 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5401 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5461 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5521 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5581 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5641 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5701 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5761 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5821 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5881 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5941 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6001 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6061 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6121 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6181 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6241 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6301 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6361 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6421 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6481 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6541 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6601 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6661 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6721 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6781 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6841 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6901 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6961 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7021 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7081 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7141 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7201 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7261 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7321 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7381 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7441 cacacaggaa acagctatga ccatgattac gccaagcttg catgcattta aatcattacg
     7501 accgagattc ccgggtaata actgatataa ttaaattgaa gc
//
