LOCUS       sequence-360116-        9877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing Murine polyomavirus 
            deltaVP1 (GAL1 promoter), VP2C-ERG20 (GAL10 promoter), and 
            VP2C-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 9877)
  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 9877)
  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 VLP NES
FEATURES             Location/Qualifiers
     source          1..9877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(268..284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(268..284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(281..303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    292..308
                     /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(316..322,323..340,341..346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    361..382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(499..516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(670..1258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(750..769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1429..2220,2221..2289))
                     /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     2052..2071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2290..2394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2462..2480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2518..2540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2640..2659
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2853..2875
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2867..2884
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2868..2884
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8297..8738
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    8297..8414
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     8709..8732
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 aaaactgtat tccctggtaa ttaagcggcc gcactagtag gactatttgc aaagggaagg
       61 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
      121 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
      181 attagagctt caatttaatt atatcagtta ttacccggga atctcggtcg taatgattta
      241 aatgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1741 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2041 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2401 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2461 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     2521 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2581 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     2641 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     2701 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     2761 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     2821 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     2881 cagtgaattc atttaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     2941 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     3001 tgcccagtat tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaaac
     3061 atgtccacaa aatcatatac cagtagagct gagactcatg caagtccggt tgcatcgaaa
     3121 cttttacgtt taatggatga aaagaagacc aatttgtgtg cttctcttga cgttcgttcg
     3181 actgatgagc tattgaaact tgttgaaacg ttgggtccat acatttgcct tttgaaaaca
     3241 cacgttgata tcttggatga tttcagttat gagggtactg tcgttccatt gaaagcattg
     3301 gcagagaaat acaagttctt gatatttgag gacagaaaat tcgccgatat cggtaacaca
     3361 gtcaaattac aatatacatc gggcgtttac cgtatcgcag aatggtctga tatcaccaac
     3421 gcccacgggg ttactggtgc tggtattgtt gctggcttga aacaaggtgc gcaagaggtc
     3481 accaaagaac caaggggatt attgatgctt gctgaattgt cttccaaggg ttctctagca
     3541 cacggtgaat atactaaggg taccgttgat attgcaaaga gtgataaaga tttcgttatt
     3601 gggttcattg ctcagaacga tatgggagga agagaagaag ggtttgattg gctaatcatg
     3661 accccaggtg taggtttaga cgacaaaggc gatgcattgg gtcagcagta cagaaccgtc
     3721 gacgaagttg taagtggtgg atcagatatc atcattgttg gcagaggact tttcgccaag
     3781 ggtagagatc ctaaggttga aggtgaaaga tacagaaatg ctggatggga agcgtaccaa
     3841 aagagaatca gcgctcccca ttaattatac aggaaactta atagaacaaa tcacatattt
     3901 aatctaatag ccacctgcat tggcacggtg caacactact tcaacttcat cttacaaaaa
     3961 gatcacgtga tctgttgtat tgggatctct agagtaacaa tatcatgaga ccttttatag
     4021 aagtggcgcc aaaactaaat gtatttgaaa atacaaaaaa cgcacaggtt tctttatata
     4081 tgtatatata tatatatata tataaaactg tacatatact gtttaaatta atctatctcg
     4141 agttataaag atgtgttata gatcattgcc ttcatgtgtt cttctaaaat tggcaagtta
     4201 tggttgtcat cgtaagagta catcataggg atcattctag ccatattcaa ggaacctatt
     4261 ctgaatgatt cagaaaaagg gtttggtgat aagcattcct tattcaaaca tttccatgct
     4321 tcggagatca ttcttattac ttcttctctg gctgaatcga cactggaacc cttatgttcc
     4381 ttcatgtaac attcgatgta agaaccatca taaccatctt ggtcttcatc tttggctgaa
     4441 cccaagtcat cagataatct caaaatggtt gcgatacttg tggaaatacc tggatagtca
     4501 tctaccaaat caactgattc ttgtgtaata ccgtcaccta acaagaaaaa catgtgggcc
     4561 aagacaacat gaacacctga agagatgata ccgttcttta agtattcttc tgcatttggc
     4621 aagtgaccag aggcgaacca ctttgcttca actaaaaatg cattgcacaa actagcccaa
     4681 gtcttttgca aggaatcaat agggttttga ccatgctttt tatagatttt gtatgcgatt
     4741 tcgttggtaa catcgtacaa agccttgaaa catactttca tgtagtctgg taattgttcg
     4801 acggcagaca attcccatct attaactgca tcagtaaaca aggtcaattc ttcaatggta
     4861 ccatagacgt cgaagatgtc atctatgatg tatataaaag aaataggctt agttaattca
     4921 actctttctt cactgaatct tggatcggtt aaagcggcca taggccacat ataccacttc
     4981 aatggttggt ttctagccaa tttcaattca ttggctaaac ctgtatcctt ccaccattga
     5041 gaaacttgga atacttcctt ttgatggata ctttgtacga tagtcaaatc catcttagct
     5101 aattcttgca aattatcgac accgtcccag tttggaccct tgcagttcat gtaggacaat
     5161 ttttgaccca taaaggtagc caatgacttg tggtatggat gtgttaaagt gttagaaacc
     5221 aatctggctt cgtgatgatt aggatctttt gtccatctac ctaacaattg actggagaaa
     5281 tcagcagctt gatctaagat gtcttcacct tctattctca attgacttgc ttcgtataag
     5341 gacatcaaac ctctaatatc tgttgacaat tctgatctga atctaccttc cttatccttg
     5401 aagttattaa agacgtcagc ggtaacattg tgaccttctt gtcttaagaa tctaaacaat
     5461 gttgaaactt cgtacaaacc atgttcaggg taaccatctg gagttgaaga tatgatttgt
     5521 tgcttttgca agaatgcacc aatttcacct ctgaaatggt aatcgatacc caatctttgc
     5581 aaagcgtcta tcataactaa accttccaaa gggttttctt caacttctct taatgctttt
     5641 ctaacatctt ccaacttttc ttcgtactta acgtataatt cgtcagaaaa tgatctgtaa
     5701 ccagtttgag ggttattgtg cttactagga ttggaaatac atgtcaaatc ttcagctata
     5761 gaccatttat gagtaattgg tacggaagga gcgattggag ttctgtaagc gttggagttt
     5821 tgtggggagt tgttagttgc gataggacct gctgcggtag ccatgagctc agaaccgccg
     5881 ccaccggagc caccgccacc agaaccacca ccacctggag tgatgtcacc atacaaaccc
     5941 aaaatcaatg gcaacatcca atctggagta actctttgat gagaaacaac ttgagcttga
     6001 taaaatttaa taacttcacc agattcatct tgaatatgag cagctggagt tggaccacca
     6061 ttacccattt tatcgatttt tgagggaata ttcaactgtt tttttttatc atgttgatgc
     6121 tctgcataat aatgcccata aatatttccg acctgctttt atatctttgc tagccaaact
     6181 aactgaacat agctacacat tattttcagc ttggctattt tgtgaacact gtatagccag
     6241 tccttcggat cacggtcaac agttgtccga gcgctttttg gaccctttcc cttatttttg
     6301 ggttaaggaa aatgacagaa aatatatcta atgagccttc gctcaacagt gctccgaagt
     6361 atagctttcc aaaaggagag gcaaagcaat ttaagaatgt atgaacaaaa taaaggggaa
     6421 aaattacccc ctctacttta ccaaacgaat actaccaata atatttacaa cttttcctta
     6481 tgattttttc actgaagcgc ttcgcaatag ttgtgagtga tatcaaaagt aacgaaatga
     6541 actccgcggc tcgtgctata ttcttgttgc taccgtccat atctttccat agattttcaa
     6601 tttttgatgt ctccatggtg gtacagagaa cttgtaaaca attcggtccc tacatgtgag
     6661 gaaattcgct gtgacacttt tatcactgaa ctccaaattt aaaaaatagc ataaaattcg
     6721 ttatacagca aatctatgtg ttgcaattaa gaactaaaag atatagagtg catattttca
     6781 agaaggatag taagctggca aaagatcttt atttgcttct cttgtaaact ttgttcaaaa
     6841 acgcagttaa gacgtcggct ttgaagccac gagactcgtc gacttgggag atcttggcct
     6901 tcaaatcctt ggcaacagac tcttcatatt cgtggtataa ctggtcgatt ttcaagtcat
     6961 tgaaaatctt tttgcatttg gcttctgcga ctgagtcctt cttaccgtaa ttttcgtcta
     7021 aagtctttct ttgttctgcg gaagcaagtt ctaatgcctt gttgattacc caagaacatt
     7081 tgttatcttg gatatctgta ccgatcttac cgatctgttc tggggtaccg aagcagtcta
     7141 agtagtcatc ttgaatttgg aaatattcac ccaatggaat caagacatct ctggcttgtt
     7201 tcaaatcctt ttcatctgtg ataccggcaa cgtacatagc caatgcgaca ggcaagtaga
     7261 aagaatagta agcagtcttg aaagtaacta tgaaggagtg cttctttagg gagaacttac
     7321 tcaagtcgac tttgtcttca ggtgcagtga ttaagtccat caattggccc aattcggttt
     7381 ggaaggtgac ttcatggaac aattcggtga tatctatgta gtatttttcg tttctgaagt
     7441 gagatttcaa aagcttgtag atagcagcct ctaacatgaa tgcgtcattg atggcaattt
     7501 ccccaacttc aggaaccttg taccaacatg gttggcctct tctggtaatg gacttgtcca
     7561 tcatatcatc ggcgaccaag aagtaagcct gcaacaactc aatgcaccaa cctagaatag
     7621 caaccttttc gtattcttct tgccccaatt gttcaacggt cttgttggag agaatagcat
     7681 acgtgtccac aacggacaaa cctctattta acttaccgcc tggagtgttg tagttcaatg
     7741 agtgggcata ccagtcacat gcttccttag gcataccgta agccaaaagc gatgcgttca
     7801 attcctctac taatttaggg aaaacgttca agaatctctc tctcctaatt tctttttctg
     7861 aagccatgga tccagaaccg ccgccaccgg agccaccgcc accagaacca ccaccacctg
     7921 gagtgatgtc accatacaaa cccaaaatca atggcaacat ccaatctgga gtaactcttt
     7981 gatgagaaac aacttgagct tgataaaatt taataacttc accagattca tcttgaatat
     8041 gagcagctgg agttggacca ccattaccca ttttatcgat ttatattgaa ttttcaaaaa
     8101 ttcttacttt tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcaata
     8161 ccaccatata catatccata tctaatctta cttatatgtt gtggaaatgt aaagagcccc
     8221 attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca
     8281 ttgctatatt gaagtacgga ttagaagccg ccgagcgggc gacagccctc cgacggaaga
     8341 ctctcctccg tgcgtcctgg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc
     8401 gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga
     8461 aaaattggca gtaacctggc cccacaaacc ttcaaatcaa cgaatcaaat taacaaccat
     8521 aggataataa tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga
     8581 tgatttttga tctattaaca gatatataaa tgcaaaagct gcataaccac tttaactaat
     8641 actttcaaca ttttcggttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa
     8701 aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactatag ctagcaaaat
     8761 ggcttcaggt gtaagtaaat gcgaaaccaa atgcacaaag gcttgcccta gaccagctcc
     8821 agttcctaag ttattgatta aaggtggtat ggaagttttg gatttggtta caggtccaga
     8881 ttctgttact gaaatcgaag catttttgaa cccaagaatg ggtcaaccac caacaccaga
     8941 atctttaact gaaggtggtc aatattacgg ttggtcaaga ggtattaatt tggctacatc
     9001 tgatactgaa gattcaccag gtaataatac attaccaact tggtctatgg caaaattgca
     9061 attaccaatg ttgaacgaag atttgacttg tgatactttg caaatgtggg aagcagtttc
     9121 agttaaaaca gaagttgttg gttctggttc tttgttggat gttcatggtt ttaataagcc
     9181 aacagatact gttaacacaa agggtatttc tactccagtt gaaggttcac aatatcatgt
     9241 ttttgctgtt ggtggtgaac cattggattt gcaaggttta gttacagatg caagaactaa
     9301 gtacaaggaa gaaggtgttg ttactattaa aacaatcact aagaaagata tggttaataa
     9361 ggatcaagtt ttgaacccaa tttcaaaggc taaattggat aaggatggca tgtacccagt
     9421 tgaaatttgg catccagatc cagctaaaaa tgaaaacaca agatacttcg gtaattacac
     9481 tggtggtact acaactccac cagttttgca attcactaac actttgacaa ctgttttgtt
     9541 agatgaaaat ggtgttggtc cattatgtaa aggtgaaggt ttgtacttat cttgcgttga
     9601 tattatgggt tggagagtta caagaaacta cgatgttcat cattggagag gtttgccaag
     9661 atacttcaag attactttaa gaaaaagatg ggttaaaaat ccatacccaa tggcttcttt
     9721 gatttcttct ttgtttaata atatgttacc acaagttcaa ggtcaaccaa tggaaggtga
     9781 aaacacacaa gttgaagaag ttagagttta cgatggtact gaaccagttc caggtgaccc
     9841 tgacatgaca agatatgtag atagattcgg taaaact
//
