LOCUS       sequence-360113-        8517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing Murine polyomavirus 
            deltaVP1 (GAL1 promoter), VP2C-GFP (GAL10 promoter), and VP2C-mRuby3
            (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 8517)
  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 8517)
  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 mRuby3 VLP
FEATURES             Location/Qualifiers
     source          1..8517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(260..276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(260..276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(273..295)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    284..300
                     /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(308..314,315..332,333..338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    353..374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(491..508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(662..1250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(742..761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1421..2212,2213..2281))
                     /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     2044..2063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2282..2386)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2454..2472
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2510..2532)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2632..2651
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2845..2867
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2859..2876
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2860..2876
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4134..4844)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     CDS             complement(5786..6496)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     promoter        6929..7370
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6929..7046
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7341..7364
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 attccctggt aattaagcgg ccgcactagt aggactattt gcaaagggaa gggatgctaa
       61 ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca
      121 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
      181 ttcaatttaa ttatatcagt tattacccgg gaatctcggt cgtaatgatt taaatgcatg
      241 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      301 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      361 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      421 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      481 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      541 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      601 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      661 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      781 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      841 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      901 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      961 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1021 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1081 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1141 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1201 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1261 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1321 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1381 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1441 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1501 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1561 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1621 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1681 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1741 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1801 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1861 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1921 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1981 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2041 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2101 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2161 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2221 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2281 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2341 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2401 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2461 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2521 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2581 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2641 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2701 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2761 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2821 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2881 tcatttaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     2941 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     3001 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac
     3061 aaaatcatat accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg
     3121 tttaatggat gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga
     3181 gctattgaaa cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga
     3241 tatcttggat gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa
     3301 atacaagttc ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt
     3361 acaatataca tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg
     3421 ggttactggt gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga
     3481 accaagggga ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga
     3541 atatactaag ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat
     3601 tgctcagaac gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg
     3661 tgtaggttta gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt
     3721 tgtaagtggt ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga
     3781 tcctaaggtt gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat
     3841 cagcgctccc cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat
     3901 agccacctgc attggcacgg tgcaacacta cttcaacttc atcttacaaa aagatcacgt
     3961 gatctgttgt attgggatct ctagagtaac aatatcatga gaccttttat agaagtggcg
     4021 ccaaaactaa atgtatttga aaatacaaaa aacgcacagg tttctttata tatgtatata
     4081 tatatatata tatatataaa actgtacata tactgtttaa attaatctat ttacttgtac
     4141 agctcgtcca tgccaccacc aaggttgctg tatttggcaa ctgccacttc gcgttgcact
     4201 acgtaggttt cattgtcact ctcctcgatc ctttccaggc ggtgatcaac ggcatggaca
     4261 ccgggcatct tgatgttccc gacggtcttt tttgacctgt aagttgtcac gaagttgcag
     4321 tgcagatggc caccaccatc aactttcagt gcgatgtcag tgtatcctct cagaccacca
     4381 tctgctggat acatcatctc tgtattaggc tcccaaccct tggtcttctt ctgcatcacg
     4441 ggaccattgg agggaaagtt tacccctctg accttgacgt tgtagacgag ctcgccatcc
     4501 tcaaggctgg tgtcctgcgt gacggtgacg actccaccat cttcgtatct cgtaactctt
     4561 tcccaagtaa aaccctcagg aaaggactgt ttaaagaaat cagggatgtc ggccgggtac
     4621 ttgataaaag tacggctgcc atacatgaac gacgtggcaa gaatgtcaaa ggcaaatggc
     4681 aggggtcctc cctcgatgac tttgatcctc atggtttgca ctccctcgta cggtctgcct
     4741 tctccttcac ctgtgcattt gaattggtgg ccgttgaccg aaccttccat gaccaccttc
     4801 atacgcatat tttccttgat cagctcttcg cccttagaca ccatagaacc gccgccaccg
     4861 gagccaccgc caccagaacc accaccacct ggagtgatgt caccatacaa acccaaaatc
     4921 aatggcaaca tccaatctgg agtaactctt tgatgagaaa caacttgagc ttgataaaat
     4981 ttaataactt caccagattc atcttgaata tgagcagctg gagttggacc accattaccc
     5041 attttatcga tttttgaggg aatattcaac tgtttttttt tatcatgttg atgctctgca
     5101 taataatgcc cataaatatt tccgacctgc ttttatatct ttgctagcca aactaactga
     5161 acatagctac acattatttt cagcttggct attttgtgaa cactgtatag ccagtccttc
     5221 ggatcacggt caacagttgt ccgagcgctt tttggaccct ttcccttatt tttgggttaa
     5281 ggaaaatgac agaaaatata tctaatgagc cttcgctcaa cagtgctccg aagtatagct
     5341 ttccaaaagg agaggcaaag caatttaaga atgtatgaac aaaataaagg ggaaaaatta
     5401 ccccctctac tttaccaaac gaatactacc aataatattt acaacttttc cttatgattt
     5461 tttcactgaa gcgcttcgca atagttgtga gtgatatcaa aagtaacgaa atgaactccg
     5521 cggctcgtgc tatattcttg ttgctaccgt ccatatcttt ccatagattt tcaatttttg
     5581 atgtctccat ggtggtacag agaacttgta aacaattcgg tccctacatg tgaggaaatt
     5641 cgctgtgaca cttttatcac tgaactccaa atttaaaaaa tagcataaaa ttcgttatac
     5701 agcaaatcta tgtgttgcaa ttaagaacta aaagatatag agtgcatatt ttcaagaagg
     5761 atagtaagct ggcaaattaa gatcttttgt acaattcatc cataccatgg gtaataccag
     5821 cagcagtaac aaattctaac aagaccatgt ggtctctctt ttcgtttgga tctttggata
     5881 aggcagattg agtggataag taatggttgt ctggtaacaa gactggacca tcaccaattg
     5941 gagtattttg ttgataatgg tcagctaatt gaacagaacc atcttcaatg ttgtgtctaa
     6001 ttttgaagtt aactttgata ccattctttt gtttgtcagc catgatgtaa acattgtgag
     6061 agttatagtt gtattccaat ttgtgaccta aaatgttacc atcttcttta aaatcaatac
     6121 cttttaattc gattctatta actaaggtat caccttcaaa cttgacttca gctctggtct
     6181 tgtagttacc gtcatctttg aaaaaaatag ttctttcttg aacataacct tctggcatgg
     6241 cagacttgaa aaagtcatgt tgtttcatat gatctgggta tctcgcaaaa cattgaacac
     6301 cataaccgaa agtagtgact aaggttggcc atggaactgg caatttacca gtagtacaaa
     6361 taaattttaa ggtcaattta ccgtaagtag catcaccttc accttcaccg gagacagaaa
     6421 atttgtgacc attaacatca ccatctaatt caaccaaaat tgggacaaca ccagtgaata
     6481 attcttcacc tttagaggat cccatagaac cgccgccacc ggagccaccg ccaccagaac
     6541 caccaccacc tggagtgatg tcaccataca aacccaaaat caatggcaac atccaatctg
     6601 gagtaactct ttgatgagaa acaacttgag cttgataaaa tttaataact tcaccagatt
     6661 catcttgaat atgagcagct ggagttggac caccattacc cattttatcg atttatattg
     6721 aattttcaaa aattcttact tttttttttg gatggacgca aagaagttta ataatcatat
     6781 tacatggcaa taccaccata tacatatcca tatctaatct tacttatatg ttgtggaaat
     6841 gtaaagagcc ccattatctt agcctaaaaa aaccttctct ttggaacttt cagtaatacg
     6901 cttaactgct cattgctata ttgaagtacg gattagaagc cgccgagcgg gcgacagccc
     6961 tccgacggaa gactctcctc cgtgcgtcct ggtcttcacc ggtcgcgttc ctgaaacgca
     7021 gatgtgcctc gcgccgcact gctccgaaca ataaagattc tacaatacta gcttttatgg
     7081 ttatgaagag gaaaaattgg cagtaacctg gccccacaaa ccttcaaatc aacgaatcaa
     7141 attaacaacc ataggataat aatgcgatta gttttttagc cttatttctg gggtaattaa
     7201 tcagcgaagc gatgattttt gatctattaa cagatatata aatgcaaaag ctgcataacc
     7261 actttaacta atactttcaa cattttcggt ttgtattact tcttattcaa atgtcataaa
     7321 agtatcaaca aaaaattgtt aatatacctc tatactttaa cgtcaaggag aaaaaactat
     7381 agctagcaaa atggcttcag gtgtaagtaa atgcgaaacc aaatgcacaa aggcttgccc
     7441 tagaccagct ccagttccta agttattgat taaaggtggt atggaagttt tggatttggt
     7501 tacaggtcca gattctgtta ctgaaatcga agcatttttg aacccaagaa tgggtcaacc
     7561 accaacacca gaatctttaa ctgaaggtgg tcaatattac ggttggtcaa gaggtattaa
     7621 tttggctaca tctgatactg aagattcacc aggtaataat acattaccaa cttggtctat
     7681 ggcaaaattg caattaccaa tgttgaacga agatttgact tgtgatactt tgcaaatgtg
     7741 ggaagcagtt tcagttaaaa cagaagttgt tggttctggt tctttgttgg atgttcatgg
     7801 ttttaataag ccaacagata ctgttaacac aaagggtatt tctactccag ttgaaggttc
     7861 acaatatcat gtttttgctg ttggtggtga accattggat ttgcaaggtt tagttacaga
     7921 tgcaagaact aagtacaagg aagaaggtgt tgttactatt aaaacaatca ctaagaaaga
     7981 tatggttaat aaggatcaag ttttgaaccc aatttcaaag gctaaattgg ataaggatgg
     8041 catgtaccca gttgaaattt ggcatccaga tccagctaaa aatgaaaaca caagatactt
     8101 cggtaattac actggtggta ctacaactcc accagttttg caattcacta acactttgac
     8161 aactgttttg ttagatgaaa atggtgttgg tccattatgt aaaggtgaag gtttgtactt
     8221 atcttgcgtt gatattatgg gttggagagt tacaagaaac tacgatgttc atcattggag
     8281 aggtttgcca agatacttca agattacttt aagaaaaaga tgggttaaaa atccataccc
     8341 aatggcttct ttgatttctt ctttgtttaa taatatgtta ccacaagttc aaggtcaacc
     8401 aatggaaggt gaaaacacac aagttgaaga agttagagtt tacgatggta ctgaaccagt
     8461 tccaggtgac cctgacatga caagatatgt agatagattc ggtaaaacta aaactgt
//
