LOCUS       sequence-360121-        8309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing ERG20(F96W-N127W) (GAL10 
            promoter) and 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, Plan MR, Peng B, Lu Z, Schenk G, Vickers CE, 
            Sainsbury F
  TITLE     Synthetic in vivo compartmentalisation improves metabolic flux and 
            modulates the product profile of promiscuous enzymes
  JOURNAL   bioRxiv 2022.11.24.517869
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: "bioRxiv 
            2022.11.24.517869"
COMMENT     Sequence Label: Coexpressed Free NES (M)
FEATURES             Location/Qualifiers
     source          1..8309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(93..534)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(99..122)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    417..534
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5539..5556)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5539..5555)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5548..5570)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5764..5783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5883..5905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5943..5961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6029..6133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6134..6202,6203..6994)
                     /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(6352..6371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7165..7753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7654..7673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7907..7924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8041..8062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8077..8082,8083..8100,8101..8107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8115..8131
                     /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     8120..8142
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8139..8155
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8139..8155
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc
       61 tttgcaaata gtcctactag ttatagtttt ttctccttga cgttaaagta tagaggtata
      121 ttaacaattt tttgttgata cttttatgac atttgaataa gaagtaatac aaaccgaaaa
      181 tgttgaaagt attagttaaa gtggttatgc agcttttgca tttatatatc tgttaataga
      241 tcaaaaatca tcgcttcgct gattaattac cccagaaata aggctaaaaa actaatcgca
      301 ttattatcct atggttgtta atttgattcg ttgatttgaa ggtttgtggg gccaggttac
      361 tgccaatttt tcctcttcat aaccataaaa gctagtattg tagaatcttt attgttcgga
      421 gcagtgcggc gcgaggcaca tctgcgtttc aggaacgcga ccggtgaaga ccaggacgca
      481 cggaggagag tcttccgtcg gagggctgtc gcccgctcgg cggcttctaa tccgtacttc
      541 aatatagcaa tgagcagtta agcgtattac tgaaagttcc aaagagaagg tttttttagg
      601 ctaagataat ggggctcttt acatttccac aacatataag taagattaga tatggatatg
      661 tatatggtgg tattgccatg taatatgatt attaaacttc tttgcgtcca tccaaaaaaa
      721 aaagtaagaa tttttgaaaa ttcaatataa ggatccaaaa tggcttcaga aaaagaaatt
      781 aggagagaga gattcttgaa cgttttccct aaattagtag aggaattgaa cgcatcgctt
      841 ttggcttacg gtatgcctaa ggaagcatgt gactggtatg cccactcatt gaactacaac
      901 actccaggcg gtaagctaaa tagaggtttg tccgttgtgg acacgtatgc tattctctcc
      961 aacaagaccg ttgaacaatt ggggcaagaa gaatacgaaa aggttgccat tctaggttgg
     1021 tgcattgagt tgttgcaggc ttactggttg gtcgccgatg atatgatgga caagtccatt
     1081 accagaagag gccaaccatg ttggtacaag gttcctgaag ttggggaaat tgccatctgg
     1141 gacgcattca tgttagaggc tgctatctac aagcttttga aatctcactt cagaaacgaa
     1201 aaatactaca tagatatcac cgaattgttc catgaggtca ccttccaaac cgaattgggc
     1261 caattgatgg acttaatcac tgcacctgaa gacaaagtcg acttgagtaa gttctcccta
     1321 aagaagcact ccttcatagt tactttcaag actgcttact attctttcta cttgcctgtc
     1381 gcattggcca tgtacgttgc cggtatcacg gatgaaaagg atttgaaaca agccagagat
     1441 gtcttgattc cattgggtga atacttccaa attcaagatg actacttaga ctgcttcggt
     1501 accccagaac agatcggtaa gatcggtaca gatatccaag ataacaaatg ttcttgggta
     1561 atcaacaagg cattagaact tgcttccgca gaacaaagaa agactttaga cgaaaattac
     1621 ggtaagaagg actcagtcgc agaagccaaa tgcaaaaaga ttttcaatga cttgaaaatt
     1681 gaacagctat accacgaata tgaagagtct attgccaagg atttgaaggc caaaatttct
     1741 caggtcgatg agtctcgtgg cttcaaagct gatgtcttaa ctgcgttctt gaacaaagtt
     1801 tacaagagaa gcaaataaag atcttttgcc agcttactat ccttcttgaa aatatgcact
     1861 ctatatcttt tagttcttaa ttgcaacaca tagatttgct gtataacgaa ttttatgcta
     1921 ttttttaaat ttggagttca gtgataaaag tgtcacagcg aatttcctca catgtaggga
     1981 ccgaattgtt tacaagttct ctgtaccacc atggagacat caaaaattga aaatctatgg
     2041 aaagatatgg acggtagcaa caagaatata gcacgagccg cggagttcat ttcgttactt
     2101 ttgatatcac tcacaactat tgcgaagcgc ttcagtgaaa aaatcataag gaaaagttgt
     2161 aaatattatt ggtagtattc gtttggtaaa gtagaggggg taatttttcc cctttatttt
     2221 gttcatacat tcttaaattg ctttgcctct ccttttggaa agctatactt cggagcactg
     2281 ttgagcgaag gctcattaga tatattttct gtcattttcc ttaacccaaa aataagggaa
     2341 agggtccaaa aagcgctcgg acaactgttg accgtgatcc gaaggactgg ctatacagtg
     2401 ttcacaaaat agccaagctg aaaataatgt gtagctatgt tcagttagtt tggctagcaa
     2461 agatataaaa gcaggtcgga aatatttatg ggcattatta tgcagagcat caacatgata
     2521 aaaaaaaaca gttgaatatt ccctcaaaag agctcaaaat ggctaccgca gcaggtccta
     2581 tcgcaactaa caactcccca caaaactcca acgcttacag aactccaatc gctccttccg
     2641 taccaattac tcataaatgg tctatagctg aagatttgac atgtatttcc aatcctagta
     2701 agcacaataa ccctcaaact ggttacagat cattttctga cgaattatac gttaagtacg
     2761 aagaaaagtt ggaagatgtt agaaaagcat taagagaagt tgaagaaaac cctttggaag
     2821 gtttagttat gatagacgct ttgcaaagat tgggtatcga ttaccatttc agaggtgaaa
     2881 ttggtgcatt cttgcaaaag caacaaatca tatcttcaac tccagatggt taccctgaac
     2941 atggtttgta cgaagtttca acattgttta gattcttaag acaagaaggt cacaatgtta
     3001 ccgctgacgt ctttaataac ttcaaggata aggaaggtag attcagatca gaattgtcaa
     3061 cagatattag aggtttgatg tccttatacg aagcaagtca attgagaata gaaggtgaag
     3121 acatcttaga tcaagctgct gatttctcca gtcaattgtt aggtagatgg acaaaagatc
     3181 ctaatcatca cgaagccaga ttggtttcta acactttaac acatccatac cacaagtcat
     3241 tggctacctt tatgggtcaa aaattgtcct acatgaactg caagggtcca aactgggacg
     3301 gtgtcgataa tttgcaagaa ttagctaaga tggatttgac tatcgtacaa agtatccatc
     3361 aaaaggaagt attccaagtt tctcaatggt ggaaggatac aggtttagcc aatgaattga
     3421 aattggctag aaaccaacca ttgaagtggt atatgtggcc tatggccgct ttaaccgatc
     3481 caagattcag tgaagaaaga gttgaattaa ctaagcctat ttcttttata tacatcatag
     3541 atgacatctt cgacgtctat ggtaccattg aagaattgac cttgtttact gatgcagtta
     3601 atagatggga attgtctgcc gtcgaacaat taccagacta catgaaagta tgtttcaagg
     3661 ctttgtacga tgttaccaac gaaatcgcat acaaaatcta taaaaagcat ggtcaaaacc
     3721 ctattgattc cttgcaaaag acttgggcta gtttgtgcaa tgcattttta gttgaagcaa
     3781 agtggttcgc ctctggtcac ttgccaaatg cagaagaata cttaaagaac ggtatcatct
     3841 cttcaggtgt tcatgttgtc ttggcccaca tgtttttctt gttaggtgac ggtattacac
     3901 aagaatcagt tgatttggta gatgactatc caggtatttc cacaagtatc gcaaccattt
     3961 tgagattatc tgatgacttg ggttcagcca aagatgaaga ccaagatggt tatgatggtt
     4021 cttacatcga atgttacatg aaggaacata agggttccag tgtcgattca gccagagaag
     4081 aagtaataag aatgatctcc gaagcatgga aatgtttgaa taaggaatgc ttatcaccaa
     4141 accctttttc tgaatcattc agaataggtt ccttgaatat ggctagaatg atccctatga
     4201 tgtactctta cgatgacaac cataacttgc caattttaga agaacacatg aaggcaatga
     4261 tctataacac atctttataa ctcgagatag attaatttaa acagtatatg tacagtttta
     4321 tatatatata tatatatata catatataaa gaaacctgtg cgttttttgt attttcaaat
     4381 acatttagtt ttggcgccac ttctataaaa ggtctcatga tattgttact ctagagatcc
     4441 caatacaaca gatcacgtga tctttttgta agatgaagtt gaagtagtgt tgcaccgtgc
     4501 caatgcaggt ggctattaga ttaaatatgt gatttgttct attaagtttc ctgtataatt
     4561 aatggggagc gctgattctc ttttggtacg cttcccatcc agcatttctg tatctttcac
     4621 cttcaacctt aggatctcta cccttggcga aaagtcctct gccaacaatg atgatatctg
     4681 atccaccact tacaacttcg tcgacggttc tgtactgctg acccaatgca tcgcctttgt
     4741 cgtctaaacc tacacctggg gtcatgatta gccaatcaaa cccttcttct cttcctccca
     4801 tatcgttctg agcaatgaac ccaataacga aatctttatc actctttgca atatcaacgg
     4861 tacccttagt atattcaccg tgtgctagag aacccttgga agacaattca gcaagcatca
     4921 ataatcccct tggttctttg gtgacctctt gcgcaccttg tttcaagcca gcaacaatac
     4981 cagcaccagt aaccccgtgg gcgttggtga tatcagacca ttctgcgata cggtaaacgc
     5041 ccgatgtata ttgtaatttg actgtgttac cgatatcggc gaattttctg tcctcaaata
     5101 tcaagaactt gtatttctct gccaatgctt tcaatggaac gacagtaccc tcataactga
     5161 aatcatccaa gatatcaacg tgtgttttca aaaggcaaat gtatggaccc aacgtttcaa
     5221 caagtttcaa tagctcatca gtcgaacgaa cgtcaagaga agcacacaaa ttggtcttct
     5281 tttcatccat taaacgtaaa agtttcgatg caaccggact tgcatgagtc tcagctctac
     5341 tggtatatga ttttgtggac atgttttatc ttcgtttcct gcaggttttt gttctgtgca
     5401 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     5461 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     5521 aaaaatttaa atgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5581 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5641 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     5701 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     5761 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5821 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5881 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5941 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6001 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6061 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6121 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6181 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6241 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6301 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6361 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6421 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6481 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6541 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6601 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6661 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6721 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6781 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7921 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7981 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     8041 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     8101 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     8161 ttacgccaag cttgcatgca tttaaatcat tacgaccgag attcccgggt aataactgat
     8221 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     8281 ttttttagtt ttgctggccg catcttctc
//
