LOCUS       sequence-360123-        8933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 (GAL1 promoter) and 
            fusion protein VP2C-ERG20(F96C)-AcNES1 (GAL10 promoter). Contains 
            uracil marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8933)
  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 8933)
  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 (G)
FEATURES             Location/Qualifiers
     source          1..8933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1186..1627)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1192..1215)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1510..1627
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(6096..6113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6096..6112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6321..6340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6440..6462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6500..6518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6586..6690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6691..6759,6760..7551)
                     /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(6909..6928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7722..8310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8211..8230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8464..8481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8598..8619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8634..8639,8640..8657,8658..8664)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8672..8688
                     /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     8677..8699
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8696..8712
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8696..8712
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atagtcctac tagtgcggcc gcttaattac cagggaatac agttttagtt ttaccgaatc
       61 tatctacata tcttgtcatg tcagggtcac ctggaactgg ttcagtacca tcgtaaactc
      121 taacttcttc aacttgtgtg ttttcacctt ccattggttg accttgaact tgtggtaaca
      181 tattattaaa caaagaagaa atcaaagaag ccattgggta tggattttta acccatcttt
      241 ttcttaaagt aatcttgaag tatcttggca aacctctcca atgatgaaca tcgtagtttc
      301 ttgtaactct ccaacccata atatcaacgc aagataagta caaaccttca cctttacata
      361 atggaccaac accattttca tctaacaaaa cagttgtcaa agtgttagtg aattgcaaaa
      421 ctggtggagt tgtagtacca ccagtgtaat taccgaagta tcttgtgttt tcatttttag
      481 ctggatctgg atgccaaatt tcaactgggt acatgccatc cttatccaat ttagcctttg
      541 aaattgggtt caaaacttga tccttattaa ccatatcttt cttagtgatt gttttaatag
      601 taacaacacc ttcttccttg tacttagttc ttgcatctgt aactaaacct tgcaaatcca
      661 atggttcacc accaacagca aaaacatgat attgtgaacc ttcaactgga gtagaaatac
      721 cctttgtgtt aacagtatct gttggcttat taaaaccatg aacatccaac aaagaaccag
      781 aaccaacaac ttctgtttta actgaaactg cttcccacat ttgcaaagta tcacaagtca
      841 aatcttcgtt caacattggt aattgcaatt ttgccataga ccaagttggt aatgtattat
      901 tacctggtga atcttcagta tcagatgtag ccaaattaat acctcttgac caaccgtaat
      961 attgaccacc ttcagttaaa gattctggtg ttggtggttg acccattctt gggttcaaaa
     1021 atgcttcgat ttcagtaaca gaatctggac ctgtaaccaa atccaaaact tccataccac
     1081 ctttaatcaa taacttagga actggagctg gtctagggca agcctttgtg catttggttt
     1141 cgcatttact tacacctgaa gccattttgc tagctatagt tttttctcct tgacgttaaa
     1201 gtatagaggt atattaacaa ttttttgttg atacttttat gacatttgaa taagaagtaa
     1261 tacaaaccga aaatgttgaa agtattagtt aaagtggtta tgcagctttt gcatttatat
     1321 atctgttaat agatcaaaaa tcatcgcttc gctgattaat taccccagaa ataaggctaa
     1381 aaaactaatc gcattattat cctatggttg ttaatttgat tcgttgattt gaaggtttgt
     1441 ggggccaggt tactgccaat ttttcctctt cataaccata aaagctagta ttgtagaatc
     1501 tttattgttc ggagcagtgc ggcgcgaggc acatctgcgt ttcaggaacg cgaccggtga
     1561 agaccaggac gcacggagga gagtcttccg tcggagggct gtcgcccgct cggcggcttc
     1621 taatccgtac ttcaatatag caatgagcag ttaagcgtat tactgaaagt tccaaagaga
     1681 aggttttttt aggctaagat aatggggctc tttacatttc cacaacatat aagtaagatt
     1741 agatatggat atgtatatgg tggtattgcc atgtaatatg attattaaac ttctttgcgt
     1801 ccatccaaaa aaaaaagtaa gaatttttga aaattcaata taaatcgata aaatgggtaa
     1861 tggtggtcca actccagctg ctcatattca agatgaatct ggtgaagtta ttaaatttta
     1921 tcaagctcaa gttgtttctc atcaaagagt tactccagat tggatgttgc cattgatttt
     1981 gggtttgtat ggtgacatca ctccaggtgg tggtggttct ggtggcggtg gctccggtgg
     2041 cggcggttct ggatccatgg cttcagaaaa agaaattagg agagagagat tcttgaacgt
     2101 tttccctaaa ttagtagagg aattgaacgc atcgcttttg gcttacggta tgcctaagga
     2161 agcatgtgac tggtatgccc actcattgaa ctacaacact ccaggcggta agctaaatag
     2221 aggtttgtcc gttgtggaca cgtatgctat tctctccaac aagaccgttg aacaattggg
     2281 gcaagaagaa tacgaaaagg ttgccattct aggttggtgc attgagttgt tgcaggctta
     2341 ctgtttggtc gccgatgata tgatggacaa gtccattacc agaagaggcc aaccatgttg
     2401 gtacaaggtt cctgaagttg gggaaattgc catcaatgac gcattcatgt tagaggctgc
     2461 tatctacaag cttttgaaat ctcacttcag aaacgaaaaa tactacatag atatcaccga
     2521 attgttccat gaggtcacct tccaaaccga attgggccaa ttgatggact taatcactgc
     2581 acctgaagac aaagtcgact tgagtaagtt ctccctaaag aagcactcct tcatagttac
     2641 tttcaagact gcttactatt ctttctactt gcctgtcgca ttggccatgt acgttgccgg
     2701 tatcacggat gaaaaggatt tgaaacaagc cagagatgtc ttgattccat tgggtgaata
     2761 cttccaaatt caagatgact acttagactg cttcggtacc ccagaacaga tcggtaagat
     2821 cggtacagat atccaagata acaaatgttc ttgggtaatc aacaaggcat tagaacttgc
     2881 ttccgcagaa caaagaaaga ctttagacga aaattacggt aagaaggact cagtcgcaga
     2941 agccaaatgc aaaaagattt tcaatgactt gaaaattgaa cagctatacc acgaatatga
     3001 agagtctatt gccaaggatt tgaaggctaa aatttctcag gtcgatgagt ctcgtggctt
     3061 caaagctgat gtcttaactg cgtttttgaa caaagtttac aagagaagca aaagatctgc
     3121 tggtggagga ggtactggtg gtgctgagct catggctacc gcagcaggtc ctatcgcaac
     3181 taacaactcc ccacaaaact ccaacgctta cagaactcca atcgctcctt ccgtaccaat
     3241 tactcataaa tggtctatag ctgaagattt gacatgtatt tccaatccta gtaagcacaa
     3301 taaccctcaa actggttaca gatcattttc tgacgaatta tacgttaagt acgaagaaaa
     3361 gttggaagat gttagaaaag cattaagaga agttgaagaa aaccctttgg aaggtttagt
     3421 tatgatagac gctttgcaaa gattgggtat cgattaccat ttcagaggtg aaattggtgc
     3481 attcttgcaa aagcaacaaa tcatatcttc aactccagat ggttaccctg aacatggttt
     3541 gtacgaagtt tcaacattgt ttagattctt aagacaagaa ggtcacaatg ttaccgctga
     3601 cgtctttaat aacttcaagg ataaggaagg tagattcaga tcagaattgt caacagatat
     3661 tagaggtttg atgtccttat acgaagcaag tcaattgaga atagaaggtg aagacatctt
     3721 agatcaagct gctgatttct ccagtcaatt gttaggtaga tggacaaaag atcctaatca
     3781 tcacgaagcc agattggttt ctaacacttt aacacatcca taccacaagt cattggctac
     3841 ctttatgggt caaaaattgt cctacatgaa ctgcaagggt ccaaactggg acggtgtcga
     3901 taatttgcaa gaattagcta agatggattt gactatcgta caaagtatcc atcaaaagga
     3961 agtattccaa gtttctcaat ggtggaagga tacaggttta gccaatgaat tgaaattggc
     4021 tagaaaccaa ccattgaagt ggtatatgtg gcctatggcc gctttaaccg atccaagatt
     4081 cagtgaagaa agagttgaat taactaagcc tatttctttt atatacatca tagatgacat
     4141 cttcgacgtc tatggtacca ttgaagaatt gaccttgttt actgatgcag ttaatagatg
     4201 ggaattgtct gccgtcgaac aattaccaga ctacatgaaa gtatgtttca aggctttgta
     4261 cgatgttacc aacgaaatcg catacaaaat ctataaaaag catggtcaaa accctattga
     4321 ttccttgcaa aagacttggg ctagtttgtg caatgcattt ttagttgaag caaagtggtt
     4381 cgcctctggt cacttgccaa atgcagaaga atacttaaag aacggtatca tctcttcagg
     4441 tgttcatgtt gtcttggccc acatgttttt cttgttaggt gacggtatta cacaagaatc
     4501 agttgatttg gtagatgact atccaggtat ttccacaagt atcgcaacca ttttgagatt
     4561 atctgatgac ttgggttcag ccaaagatga agaccaagat ggttatgatg gttcttacat
     4621 cgaatgttac atgaaggaac ataagggttc cagtgtcgat tcagccagag aagaagtaat
     4681 aagaatgatc tccgaagcat ggaaatgttt gaataaggaa tgcttatcac caaacccttt
     4741 ttctgaatca ttcagaatag gttccttgaa tatggctaga atgatcccta tgatgtactc
     4801 ttacgatgac aaccataact tgccaatttt agaagaacac atgaaggcaa tgatctataa
     4861 cacatcttta taactcgaga tagattaatt taaacagcat atataaagaa acctgtgcgt
     4921 tttttgtatt ttcaaataca tttagttttg gcgccacttc tataaaaggt ctcatgatat
     4981 tgttactcta gagatcccaa tacaacagat cacgtgatct ttttgtaaga tgaagttgaa
     5041 gtagtgttgc accgtgccaa tgcaggtggc tattagatta aatatgtgat ttgttctatt
     5101 aagtttcctg tataattaat ggggagcgct gattctcttt tggtacgctt cccatccagc
     5161 atttctgtat ctttcacctt caaccttagg atctctaccc ttggcgaaaa gtcctctgcc
     5221 aacaatgatg atatctgatc caccacttac aacttcgtcg acggttctgt actgctgacc
     5281 caatgcatcg cctttgtcgt ctaaacctac acctggggtc atgattagcc aatcaaaccc
     5341 ttcttctctt cctcccatat cgttctgagc aatgaaccca ataacgaaat ctttatcact
     5401 ctttgcaata tcaacggtac ccttagtata ttcaccgtgt gctagagaac ccttggaaga
     5461 caattcagca agcatcaata atccccttgg ttctttggtg acctcttgcg caccttgttt
     5521 caagccagca acaataccag caccagtaac cccgtgggcg ttggtgatat cagaccattc
     5581 tgcgatacgg taaacgcccg atgtatattg taatttgact gtgttaccga tatcggcgaa
     5641 ttttctgtcc tcaaatatca agaacttgta tttctctgcc aatgctttca atggaacgac
     5701 agtaccctca taactgaaat catccaagat atcaacgtgt gttttcaaaa ggcaaatgta
     5761 tggacccaac gtttcaacaa gtttcaatag ctcatcagtc gaacgaacgt caagagaagc
     5821 acacaaattg gtcttctttt catccattaa acgtaaaagt ttcgatgcaa ccggacttgc
     5881 atgagtctca gctctactgg tatatgattt tgtggacatg ttttatcttc gtttcctgca
     5941 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     6001 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     6061 ggagattacc gaatcaaaaa aatttaaatg aattcactgg ccgtcgtttt acaacgtcgt
     6121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     6181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     6241 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     6301 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     6361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6481 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7741 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7801 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7861 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7921 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7981 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     8041 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     8101 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8161 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8221 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8281 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8341 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8401 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8461 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8521 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8581 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8641 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8701 aacagctatg accatgatta cgccaagctt gcatgcattt aaatcattac gaccgagatt
     8761 cccgggtaat aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc
     8821 atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca
     8881 gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg caa
//
