LOCUS       sequence-360436-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20-AcNES1, 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 7605)
  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 7605)
  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
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(131..148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(302..890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(382..401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1061..1852,1853..1921))
                     /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     1684..1703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1922..2026)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2094..2112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2150..2172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2272..2291
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2485..2507
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2499..2516
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2500..2516
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6817..7258
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6817..6934
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7229..7252
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(7505..7521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7505..7521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7518..7540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7529..7545
                     /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(7553..7559,7560..7577,7578..7583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(7598..7605,1..14)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
       61 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      121 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      181 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      241 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      301 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      361 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      421 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      481 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      541 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      601 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      661 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      721 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      781 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      841 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      901 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      961 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1021 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1081 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1141 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1201 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1261 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1321 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1381 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1441 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1501 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1561 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1621 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1681 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1741 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1801 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1861 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1921 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1981 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2041 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2101 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2161 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2221 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2281 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2341 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2401 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2461 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2521 tcatttaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     2581 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     2641 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac
     2701 aaaatcatat accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg
     2761 tttaatggat gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga
     2821 gctattgaaa cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga
     2881 tatcttggat gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa
     2941 atacaagttc ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt
     3001 acaatataca tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg
     3061 ggttactggt gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga
     3121 accaagggga ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga
     3181 atatactaag ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat
     3241 tgctcagaac gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg
     3301 tgtaggttta gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt
     3361 tgtaagtggt ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga
     3421 tcctaaggtt gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat
     3481 cagcgctccc cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat
     3541 agccacctgc attggcacgg tgcaacacta cttcaacttc atcttacaaa aagatcacgt
     3601 gatctgttgt attgggatct ctagagtaac aatatcatga gaccttttat agaagtggcg
     3661 ccaaaactaa atgtatttga aaatacaaaa aacgcacagg tttctttata tatgtatata
     3721 tatatatata tatataaaac tgtacatata ctgtttaaat taatctatct cgagttataa
     3781 agatgtgtta tagatcattg ccttcatgtg ttcttctaaa attggcaagt tatggttgtc
     3841 atcgtaagag tacatcatag ggatcattct agccatattc aaggaaccta ttctgaatga
     3901 ttcagaaaaa gggtttggtg ataagcattc cttattcaaa catttccatg cttcggagat
     3961 cattcttatt acttcttctc tggctgaatc gacactggaa cccttatgtt ccttcatgta
     4021 acattcgatg taagaaccat cataaccatc ttggtcttca tctttggctg aacccaagtc
     4081 atcagataat ctcaaaatgg ttgcgatact tgtggaaata cctggatagt catctaccaa
     4141 atcaactgat tcttgtgtaa taccgtcacc taacaagaaa aacatgtggg ccaagacaac
     4201 atgaacacct gaagagatga taccgttctt taagtattct tctgcatttg gcaagtgacc
     4261 agaggcgaac cactttgctt caactaaaaa tgcattgcac aaactagccc aagtcttttg
     4321 caaggaatca atagggtttt gaccatgctt tttatagatt ttgtatgcga tttcgttggt
     4381 aacatcgtac aaagccttga aacatacttt catgtagtct ggtaattgtt cgacggcaga
     4441 caattcccat ctattaactg catcagtaaa caaggtcaat tcttcaatgg taccatagac
     4501 gtcgaagatg tcatctatga tgtatataaa agaaataggc ttagttaatt caactctttc
     4561 ttcactgaat cttggatcgg ttaaagcggc cataggccac atataccact tcaatggttg
     4621 gtttctagcc aatttcaatt cattggctaa acctgtatcc ttccaccatt gagaaacttg
     4681 gaatacttcc ttttgatgga tactttgtac gatagtcaaa tccatcttag ctaattcttg
     4741 caaattatcg acaccgtccc agtttggacc cttgcagttc atgtaggaca atttttgacc
     4801 cataaaggta gccaatgact tgtggtatgg atgtgttaaa gtgttagaaa ccaatctggc
     4861 ttcgtgatga ttaggatctt ttgtccatct acctaacaat tgactggaga aatcagcagc
     4921 ttgatctaag atgtcttcac cttctattct caattgactt gcttcgtata aggacatcaa
     4981 acctctaata tctgttgaca attctgatct gaatctacct tccttatcct tgaagttatt
     5041 aaagacgtca gcggtaacat tgtgaccttc ttgtcttaag aatctaaaca atgttgaaac
     5101 ttcgtacaaa ccatgttcag ggtaaccatc tggagttgaa gatatgattt gttgcttttg
     5161 caagaatgca ccaatttcac ctctgaaatg gtaatcgata cccaatcttt gcaaagcgtc
     5221 tatcataact aaaccttcca aagggttttc ttcaacttct cttaatgctt ttctaacatc
     5281 ttccaacttt tcttcgtact taacgtataa ttcgtcagaa aatgatctgt aaccagtttg
     5341 agggttattg tgcttactag gattggaaat acatgtcaaa tcttcagcta tagaccattt
     5401 atgagtaatt ggtacggaag gagcgattgg agttctgtaa gcgttggagt tttgtgggga
     5461 gttgttagtt gcgataggac ctgctgcggt agccatgagc tcagcaccac cagtacctcc
     5521 tccaccagca gatcttttgc ttctcttgta aactttgttc aaaaacgcag ttaagacgtc
     5581 ggctttgaag ccacgagact cgtcgacttg ggagatcttg gccttcaaat ccttggcaac
     5641 agactcttca tattcgtggt ataactggtc gattttcaag tcattgaaaa tctttttgca
     5701 tttggcttct gcgactgagt ccttcttacc gtaattttcg tctaaagtct ttctttgttc
     5761 tgcggaagca agttctaatg ccttgttgat tacccaagaa catttgttat cttggatatc
     5821 tgtaccgatc ttaccgatct gttctggggt accgaagcag tctaagtagt catcttgaat
     5881 ttggaaatat tcacccaatg gaatcaagac atctctggct tgtttcaaat ccttttcatc
     5941 tgtgataccg gcaacgtaca tagccaatgc gacaggcaag tagaaagaat agtaagcagt
     6001 cttgaaagta actatgaagg agtgcttctt tagggagaac ttactcaagt cgactttgtc
     6061 ttcaggtgca gtgattaagt ccatcaattg gcccaattcg gtttggaagg tgacttcatg
     6121 gaacaattcg gtgatatcta tgtagtattt ttcgtttctg aagtgagatt tcaaaagctt
     6181 gtagatagca gcctctaaca tgaatgcgtc attgatggca atttccccaa cttcaggaac
     6241 cttgtaccaa catggttggc ctcttctggt aatggacttg tccatcatat catcggcgac
     6301 caagaagtaa gcctgcaaca actcaatgca ccaacctaga atagcaacct tttcgtattc
     6361 ttcttgcccc aattgttcaa cggtcttgtt ggagagaata gcatacgtgt ccacaacgga
     6421 caaacctcta tttaacttac cgcctggagt gttgtagttc aatgagtggg cataccagtc
     6481 acatgcttcc ttaggcatac cgtaagccaa aagcgatgcg ttcaattcct ctactaattt
     6541 agggaaaacg ttcaagaatc tctctctcct aatttctttt tctgaagcca ttttggatcc
     6601 ttatattgaa ttttcaaaaa ttcttacttt tttttttgga tggacgcaaa gaagtttaat
     6661 aatcatatta catggcaata ccaccatata catatccata tctaatctta cttatatgtt
     6721 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
     6781 gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggc
     6841 gacagccctc cgacggaaga ctctcctccg tgcgtcctgg tcttcaccgg tcgcgttcct
     6901 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
     6961 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatcaa
     7021 cgaatcaaat taacaaccat aggataataa tgcgattagt tttttagcct tatttctggg
     7081 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaagct
     7141 gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat
     7201 gtcataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
     7261 aaaactataa ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     7321 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     7381 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     7441 tcagttatta cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat
     7501 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7561 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtga
//
