LOCUS       sequence-360114-        8969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing Murine polyomavirus 
            deltaVP1 (GAL1 promoter) and fusion protein VP2C-ERG20-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 8969)
  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 8969)
  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 VLP NES
FEATURES             Location/Qualifiers
     source          1..8969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(257..273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(257..273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(270..292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    281..297
                     /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(305..311,312..329,330..335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(488..505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(659..1247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(739..758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1418..2209,2210..2278))
                     /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     2041..2060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2279..2383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2451..2469
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2507..2529)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2629..2648
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2842..2864
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2856..2873
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2857..2873
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7378..7819
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7378..7495
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7790..7813
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 ccctggtaat taagcggccg cactagtagg actatttgca aagggaaggg atgctaaggt
       61 agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca
      121 aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc
      181 aatttaatta tatcagttat tacccgggaa tctcggtcgt aatgatttaa atgcatgcaa
      241 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      301 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      361 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      421 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      481 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      541 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      601 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      661 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      721 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      781 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      841 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      901 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      961 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1021 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1081 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1141 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1201 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1261 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1321 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1381 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1441 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1501 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1561 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1621 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1681 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1741 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1801 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1861 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1921 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1981 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2041 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2101 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2161 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2221 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2281 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2341 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2401 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     2461 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2521 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2581 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2641 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2701 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2761 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2821 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattca
     2881 tttaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     2941 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     3001 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaaaca tgtccacaaa
     3061 atcatatacc agtagagctg agactcatgc aagtccggtt gcatcgaaac ttttacgttt
     3121 aatggatgaa aagaagacca atttgtgtgc ttctcttgac gttcgttcga ctgatgagct
     3181 attgaaactt gttgaaacgt tgggtccata catttgcctt ttgaaaacac acgttgatat
     3241 cttggatgat ttcagttatg agggtactgt cgttccattg aaagcattgg cagagaaata
     3301 caagttcttg atatttgagg acagaaaatt cgccgatatc ggtaacacag tcaaattaca
     3361 atatacatcg ggcgtttacc gtatcgcaga atggtctgat atcaccaacg cccacggggt
     3421 tactggtgct ggtattgttg ctggcttgaa acaaggtgcg caagaggtca ccaaagaacc
     3481 aaggggatta ttgatgcttg ctgaattgtc ttccaagggt tctctagcac acggtgaata
     3541 tactaagggt accgttgata ttgcaaagag tgataaagat ttcgttattg ggttcattgc
     3601 tcagaacgat atgggaggaa gagaagaagg gtttgattgg ctaatcatga ccccaggtgt
     3661 aggtttagac gacaaaggcg atgcattggg tcagcagtac agaaccgtcg acgaagttgt
     3721 aagtggtgga tcagatatca tcattgttgg cagaggactt ttcgccaagg gtagagatcc
     3781 taaggttgaa ggtgaaagat acagaaatgc tggatgggaa gcgtaccaaa agagaatcag
     3841 cgctccccat taattataca ggaaacttaa tagaacaaat cacatattta atctaatagc
     3901 cacctgcatt ggcacggtgc aacactactt caacttcatc ttacaaaaag atcacgtgat
     3961 ctgttgtatt gggatctcta gagtaacaat atcatgagac cttttataga agtggcgcca
     4021 aaactaaatg tatttgaaaa tacaaaaaac gcacaggttt ctttatatat gtatatatat
     4081 atatatatat ataaaactgt acatatactg tttaaattaa tctatctcga gttataaaga
     4141 tgtgttatag atcattgcct tcatgtgttc ttctaaaatt ggcaagttat ggttgtcatc
     4201 gtaagagtac atcataggga tcattctagc catattcaag gaacctattc tgaatgattc
     4261 agaaaaaggg tttggtgata agcattcctt attcaaacat ttccatgctt cggagatcat
     4321 tcttattact tcttctctgg ctgaatcgac actggaaccc ttatgttcct tcatgtaaca
     4381 ttcgatgtaa gaaccatcat aaccatcttg gtcttcatct ttggctgaac ccaagtcatc
     4441 agataatctc aaaatggttg cgatacttgt ggaaatacct ggatagtcat ctaccaaatc
     4501 aactgattct tgtgtaatac cgtcacctaa caagaaaaac atgtgggcca agacaacatg
     4561 aacacctgaa gagatgatac cgttctttaa gtattcttct gcatttggca agtgaccaga
     4621 ggcgaaccac tttgcttcaa ctaaaaatgc attgcacaaa ctagcccaag tcttttgcaa
     4681 ggaatcaata gggttttgac catgcttttt atagattttg tatgcgattt cgttggtaac
     4741 atcgtacaaa gccttgaaac atactttcat gtagtctggt aattgttcga cggcagacaa
     4801 ttcccatcta ttaactgcat cagtaaacaa ggtcaattct tcaatggtac catagacgtc
     4861 gaagatgtca tctatgatgt atataaaaga aataggctta gttaattcaa ctctttcttc
     4921 actgaatctt ggatcggtta aagcggccat aggccacata taccacttca atggttggtt
     4981 tctagccaat ttcaattcat tggctaaacc tgtatccttc caccattgag aaacttggaa
     5041 tacttccttt tgatggatac tttgtacgat agtcaaatcc atcttagcta attcttgcaa
     5101 attatcgaca ccgtcccagt ttggaccctt gcagttcatg taggacaatt tttgacccat
     5161 aaaggtagcc aatgacttgt ggtatggatg tgttaaagtg ttagaaacca atctggcttc
     5221 gtgatgatta ggatcttttg tccatctacc taacaattga ctggagaaat cagcagcttg
     5281 atctaagatg tcttcacctt ctattctcaa ttgacttgct tcgtataagg acatcaaacc
     5341 tctaatatct gttgacaatt ctgatctgaa tctaccttcc ttatccttga agttattaaa
     5401 gacgtcagcg gtaacattgt gaccttcttg tcttaagaat ctaaacaatg ttgaaacttc
     5461 gtacaaacca tgttcagggt aaccatctgg agttgaagat atgatttgtt gcttttgcaa
     5521 gaatgcacca atttcacctc tgaaatggta atcgataccc aatctttgca aagcgtctat
     5581 cataactaaa ccttccaaag ggttttcttc aacttctctt aatgcttttc taacatcttc
     5641 caacttttct tcgtacttaa cgtataattc gtcagaaaat gatctgtaac cagtttgagg
     5701 gttattgtgc ttactaggat tggaaataca tgtcaaatct tcagctatag accatttatg
     5761 agtaattggt acggaaggag cgattggagt tctgtaagcg ttggagtttt gtggggagtt
     5821 gttagttgcg ataggacctg ctgcggtagc catgagctca gcaccaccag tacctcctcc
     5881 accagcagat cttttgcttc tcttgtaaac tttgttcaaa aacgcagtta agacgtcggc
     5941 tttgaagcca cgagactcgt cgacttggga gatcttggcc ttcaaatcct tggcaacaga
     6001 ctcttcatat tcgtggtata actggtcgat tttcaagtca ttgaaaatct ttttgcattt
     6061 ggcttctgcg actgagtcct tcttaccgta attttcgtct aaagtctttc tttgttctgc
     6121 ggaagcaagt tctaatgcct tgttgattac ccaagaacat ttgttatctt ggatatctgt
     6181 accgatctta ccgatctgtt ctggggtacc gaagcagtct aagtagtcat cttgaatttg
     6241 gaaatattca cccaatggaa tcaagacatc tctggcttgt ttcaaatcct tttcatctgt
     6301 gataccggca acgtacatag ccaatgcgac aggcaagtag aaagaatagt aagcagtctt
     6361 gaaagtaact atgaaggagt gcttctttag ggagaactta ctcaagtcga ctttgtcttc
     6421 aggtgcagtg attaagtcca tcaattggcc caattcggtt tggaaggtga cttcatggaa
     6481 caattcggtg atatctatgt agtatttttc gtttctgaag tgagatttca aaagcttgta
     6541 gatagcagcc tctaacatga atgcgtcatt gatggcaatt tccccaactt caggaacctt
     6601 gtaccaacat ggttggcctc ttctggtaat ggacttgtcc atcatatcat cggcgaccaa
     6661 gaagtaagcc tgcaacaact caatgcacca acctagaata gcaacctttt cgtattcttc
     6721 ttgccccaat tgttcaacgg tcttgttgga gagaatagca tacgtgtcca caacggacaa
     6781 acctctattt aacttaccgc ctggagtgtt gtagttcaat gagtgggcat accagtcaca
     6841 tgcttcctta ggcataccgt aagccaaaag cgatgcgttc aattcctcta ctaatttagg
     6901 gaaaacgttc aagaatctct ctctcctaat ttctttttct gaagccatgg atccagaacc
     6961 gccgccaccg gagccaccgc caccagaacc accaccacct ggagtgatgt caccatacaa
     7021 acccaaaatc aatggcaaca tccaatctgg agtaactctt tgatgagaaa caacttgagc
     7081 ttgataaaat ttaataactt caccagattc atcttgaata tgagcagctg gagttggacc
     7141 accattaccc attttatcga tttatattga attttcaaaa attcttactt ttttttttgg
     7201 atggacgcaa agaagtttaa taatcatatt acatggcaat accaccatat acatatccat
     7261 atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta gcctaaaaaa
     7321 accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat tgaagtacgg
     7381 attagaagcc gccgagcggg cgacagccct ccgacggaag actctcctcc gtgcgtcctg
     7441 gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa
     7501 taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc agtaacctgg
     7561 ccccacaaac cttcaaatca acgaatcaaa ttaacaacca taggataata atgcgattag
     7621 ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg atctattaac
     7681 agatatataa atgcaaaagc tgcataacca ctttaactaa tactttcaac attttcggtt
     7741 tgtattactt cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta atatacctct
     7801 atactttaac gtcaaggaga aaaaactata gctagcaaaa tggcttcagg tgtaagtaaa
     7861 tgcgaaacca aatgcacaaa ggcttgccct agaccagctc cagttcctaa gttattgatt
     7921 aaaggtggta tggaagtttt ggatttggtt acaggtccag attctgttac tgaaatcgaa
     7981 gcatttttga acccaagaat gggtcaacca ccaacaccag aatctttaac tgaaggtggt
     8041 caatattacg gttggtcaag aggtattaat ttggctacat ctgatactga agattcacca
     8101 ggtaataata cattaccaac ttggtctatg gcaaaattgc aattaccaat gttgaacgaa
     8161 gatttgactt gtgatacttt gcaaatgtgg gaagcagttt cagttaaaac agaagttgtt
     8221 ggttctggtt ctttgttgga tgttcatggt tttaataagc caacagatac tgttaacaca
     8281 aagggtattt ctactccagt tgaaggttca caatatcatg tttttgctgt tggtggtgaa
     8341 ccattggatt tgcaaggttt agttacagat gcaagaacta agtacaagga agaaggtgtt
     8401 gttactatta aaacaatcac taagaaagat atggttaata aggatcaagt tttgaaccca
     8461 atttcaaagg ctaaattgga taaggatggc atgtacccag ttgaaatttg gcatccagat
     8521 ccagctaaaa atgaaaacac aagatacttc ggtaattaca ctggtggtac tacaactcca
     8581 ccagttttgc aattcactaa cactttgaca actgttttgt tagatgaaaa tggtgttggt
     8641 ccattatgta aaggtgaagg tttgtactta tcttgcgttg atattatggg ttggagagtt
     8701 acaagaaact acgatgttca tcattggaga ggtttgccaa gatacttcaa gattacttta
     8761 agaaaaagat gggttaaaaa tccataccca atggcttctt tgatttcttc tttgtttaat
     8821 aatatgttac cacaagttca aggtcaacca atggaaggtg aaaacacaca agttgaagaa
     8881 gttagagttt acgatggtac tgaaccagtt ccaggtgacc ctgacatgac aagatatgta
     8941 gatagattcg gtaaaactaa aactgtatt
//
