LOCUS       sequence-360120-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20(F96W-N127W)-AcNES1 (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, 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 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: "bioRxiv 
            2022.11.24.517869"
COMMENT     Sequence Label: Linked Free NES (M)
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(75..516)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(81..104)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    399..516
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4817..4834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4817..4833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4826..4848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5042..5061)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5161..5183
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5221..5239)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5307..5411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5412..5480,5481..6272)
                     /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(5630..5649)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6443..7031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6932..6951
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7185..7202
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7319..7340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7355..7360,7361..7378,7379..7385)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7393..7409
                     /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     7398..7420
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7417..7433
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7417..7433
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctact
       61 agttatagtt ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga
      121 tacttttatg acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta
      181 aagtggttat gcagcttttg catttatata tctgttaata gatcaaaaat catcgcttcg
      241 ctgattaatt accccagaaa taaggctaaa aaactaatcg cattattatc ctatggttgt
      301 taatttgatt cgttgatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc
      361 ataaccataa aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca
      421 catctgcgtt tcaggaacgc gaccggtgaa gaccaggacg cacggaggag agtcttccgt
      481 cggagggctg tcgcccgctc ggcggcttct aatccgtact tcaatatagc aatgagcagt
      541 taagcgtatt actgaaagtt ccaaagagaa ggttttttta ggctaagata atggggcttt
      601 ttacatttcc acaacatata agtaagatta gatatggata tgtatatggt ggtattgcca
      661 tgtaatatga ttattaaact tctttgcgtc catccaaaaa aaaaagtaag aatttttgaa
      721 aattcaatat aaggatccaa aatggcttca gaaaaagaaa ttaggagaga gagattcttg
      781 aacgttttcc ctaaattagt agaggaattg aacgcatcgc ttttggctta cggtatgcct
      841 aaggaagcat gtgactggta tgcccactca ttgaactaca acactccagg cggtaagcta
      901 aatagaggtt tgtccgttgt ggacacgtat gctattctct ccaacaagac cgttgaacaa
      961 ttggggcaag aagaatacga aaaggttgcc attctaggtt ggtgcattga gttgttgcag
     1021 gcttactggt tggtcgccga tgatatgatg gacaagtcca ttaccagaag aggccaacca
     1081 tgttggtaca aggttcctga agttggggaa attgccatct gggacgcatt catgttagag
     1141 gctgctatct acaagctttt gaaatctcac ttcagaaacg aaaaatacta catagatatc
     1201 accgaattgt tccatgaggt caccttccaa accgaattgg gccaattgat ggacttaatc
     1261 actgcacctg aagacaaagt cgacttgagt aagttctccc taaagaagca ctccttcata
     1321 gttactttca agactgctta ctattctttc tacttgcctg tcgcattggc catgtacgtt
     1381 gccggtatca cggatgaaaa ggatttgaaa caagccagag atgtcttgat tccattgggt
     1441 gaatacttcc aaattcaaga tgactactta gactgcttcg gtaccccaga acagatcggt
     1501 aagatcggta cagatatcca agataacaaa tgttcttggg taatcaacaa ggcattagaa
     1561 cttgcttccg cagaacaaag aaagacttta gacgaaaatt acggtaagaa ggactcagtc
     1621 gcagaagcca aatgcaaaaa gattttcaat gacttgaaaa ttgaacagct ataccacgaa
     1681 tatgaagagt ctattgccaa ggatttgaag gccaaaattt ctcaggtcga tgagtctcgt
     1741 ggcttcaaag ctgatgtctt aactgcgttc ttgaacaaag tttacaagag aagcaaaaga
     1801 tctgctggtg gaggaggtac tggtggtgct gagctcatgg ctaccgcagc aggtcctatc
     1861 gcaactaaca actccccaca aaactccaac gcttacagaa ctccaatcgc tccttccgta
     1921 ccaattactc ataaatggtc tatagctgaa gatttgacat gtatttccaa tcctagtaag
     1981 cacaataacc ctcaaactgg ttacagatca ttttctgacg aattatacgt taagtacgaa
     2041 gaaaagttgg aagatgttag aaaagcatta agagaagttg aagaaaaccc tttggaaggt
     2101 ttagttatga tagacgcttt gcaaagattg ggtatcgatt accatttcag aggtgaaatt
     2161 ggtgcattct tgcaaaagca acaaatcata tcttcaactc cagatggtta ccctgaacat
     2221 ggtttgtacg aagtttcaac attgtttaga ttcttaagac aagaaggtca caatgttacc
     2281 gctgacgtct ttaataactt caaggataag gaaggtagat tcagatcaga attgtcaaca
     2341 gatattagag gtttgatgtc cttatacgaa gcaagtcaat tgagaataga aggtgaagac
     2401 atcttagatc aagctgctga tttctccagt caattgttag gtagatggac aaaagatcct
     2461 aatcatcacg aagccagatt ggtttctaac actttaacac atccatacca caagtcattg
     2521 gctaccttta tgggtcaaaa attgtcctac atgaactgca agggtccaaa ctgggacggt
     2581 gtcgataatt tgcaagaatt agctaagatg gatttgacta tcgtacaaag tatccatcaa
     2641 aaggaagtat tccaagtttc tcaatggtgg aaggatacag gtttagccaa tgaattgaaa
     2701 ttggctagaa accaaccatt gaagtggtat atgtggccta tggccgcttt aaccgatcca
     2761 agattcagtg aagaaagagt tgaattaact aagcctattt cttttatata catcatagat
     2821 gacatcttcg acgtctatgg taccattgaa gaattgacct tgtttactga tgcagttaat
     2881 agatgggaat tgtctgccgt cgaacaatta ccagactaca tgaaagtatg tttcaaggct
     2941 ttgtacgatg ttaccaacga aatcgcatac aaaatctata aaaagcatgg tcaaaaccct
     3001 attgattcct tgcaaaagac ttgggctagt ttgtgcaatg catttttagt tgaagcaaag
     3061 tggttcgcct ctggtcactt gccaaatgca gaagaatact taaagaacgg tatcatctct
     3121 tcaggtgttc atgttgtctt ggcccacatg tttttcttgt taggtgacgg tattacacaa
     3181 gaatcagttg atttggtaga tgactatcca ggtatttcca caagtatcgc aaccattttg
     3241 agattatctg atgacttggg ttcagccaaa gatgaagacc aagatggtta tgatggttct
     3301 tacatcgaat gttacatgaa ggaacataag ggttccagtg tcgattcagc cagagaagaa
     3361 gtaataagaa tgatctccga agcatggaaa tgtttgaata aggaatgctt atcaccaaac
     3421 cctttttctg aatcattcag aataggttcc ttgaatatgg ctagaatgat ccctatgatg
     3481 tactcttacg atgacaacca taacttgcca attttagaag aacacatgaa ggcaatgatc
     3541 tataacacat ctttataact cgagatagat taatttaaac agtatatgta cagttttata
     3601 tatatatata tatatataca tatataaaga aacctgtgcg ttttttgtat tttcaaatac
     3661 atttagtttt ggcgccactt ctataaaagg tctcatgata ttgttactct agagatccca
     3721 atacaacaga tcacgtgatc tttttgtaag atgaagttga agtagtgttg caccgtgcca
     3781 atgcaggtgg ctattagatt aaatatgtga tttgttctat taagtttcct gtataattaa
     3841 tggggagcgc tgattctctt ttggtacgct tcccatccag catttctgta tctttcacct
     3901 tcaaccttag gatctctacc cttggcgaaa agtcctctgc caacaatgat gatatctgat
     3961 ccaccactta caacttcgtc gacggttctg tactgctgac ccaatgcatc gcctttgtcg
     4021 tctaaaccta cacctggggt catgattagc caatcaaacc cttcttctct tcctcccata
     4081 tcgttctgag caatgaaccc aataacgaaa tctttatcac tctttgcaat atcaacggta
     4141 cccttagtat attcaccgtg tgctagagaa cccttggaag acaattcagc aagcatcaat
     4201 aatccccttg gttctttggt gacctcttgc gcaccttgtt tcaagccagc aacaatacca
     4261 gcaccagtaa ccccgtgggc gttggtgata tcagaccatt ctgcgatacg gtaaacgccc
     4321 gatgtatatt gtaatttgac tgtgttaccg atatcggcga attttctgtc ctcaaatatc
     4381 aagaacttgt atttctctgc caatgctttc aatggaacga cagtaccctc ataactgaaa
     4441 tcatccaaga tatcaacgtg tgttttcaaa aggcaaatgt atggacccaa cgtttcaaca
     4501 agtttcaata gctcatcagt cgaacgaacg tcaagagaag cacacaaatt ggtcttcttt
     4561 tcatccatta aacgtaaaag tttcgatgca accggacttg catgagtctc agctctactg
     4621 gtatatgatt ttgtggacat gttttatctt cgtttcctgc aggtttttgt tctgtgcagt
     4681 tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc
     4741 aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa
     4801 aaatttaaat gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4861 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4921 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     4981 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     5041 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5101 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5161 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5221 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5281 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5341 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5401 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5461 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5521 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5581 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5641 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5701 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5761 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5821 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5881 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5941 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6001 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6061 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6121 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6181 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6241 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6301 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6361 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6421 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6481 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6541 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6601 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6661 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6721 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6781 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6841 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6901 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6961 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7021 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7081 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7141 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7201 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7261 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7321 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7381 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7441 acgccaagct tgcatgcatt taaatcatta cgaccgagat tcccgggtaa taactgatat
     7501 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     7561 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcct
//
