LOCUS       sequence-360432-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 with the 63 C-ter 
            residues truncated (GAL1 promoter) and destabilised GFP tagged with 
            VP2C (GAL10 promoter). Contains uracil marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  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 6791)
  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: deltaVP1d63C + VP2C-GFPdeg
FEATURES             Location/Qualifiers
     source          1..6791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(484..1194)
                     /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        1627..2068
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1627..1744
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2039..2062
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3286..3302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3286..3302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3299..3321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3310..3326
                     /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(3334..3340,3341..3358,3359..3364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3379..3400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4447..5238,5239..5307))
                     /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     5070..5089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5308..5412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5480..5498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5536..5558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5658..5677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5871..5893
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5885..5902
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5886..5902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgccaagggt agagatccta aggttgaagg tgaaagatac agaaatgctg gatgggaagc
       61 gtaccaaaag agaatcagcg ctccccatta attatacagg aaacttaata gaacaaatca
      121 catatttaat ctaatagcca cctgcattgg cacggtgcaa cactacttca acttcatctt
      181 acaaaaagat cacgtgatct gttgtattgg gatctctaga gtaacaatat catgagacct
      241 tttatagaag tggcgccaaa actaaatgta tttgaaaata caaaaaacgc acaggtttct
      301 ttatatatgt atatatatat atatatatat aaaactgtac atatactgtt taaattaatc
      361 tatttaaaca ttaattcttg cagatgcaca agcagctgga tgcctatcca tacctgattc
      421 ttgtgcacat gacattggca aagttccatc atcttgctct tcaacttcag gaggaaaaga
      481 tcttttgtac aattcatcca taccatgggt aataccagca gcagtaacaa attctaacaa
      541 gaccatgtgg tctctctttt cgtttggatc tttggataag gcagattgag tggataagta
      601 atggttgtct ggtaacaaga ctggaccatc accaattgga gtattttgtt gataatggtc
      661 agctaattga acagaaccat cttcaatgtt gtgtctaatt ttgaagttaa ctttgatacc
      721 attcttttgt ttgtcagcca tgatgtaaac attgtgagag ttatagttgt attccaattt
      781 gtgacctaaa atgttaccat cttctttaaa atcaatacct tttaattcga ttctattaac
      841 taaggtatca ccttcaaact tgacttcagc tctggtcttg tagttaccgt catctttgaa
      901 aaaaatagtt ctttcttgaa cataaccttc tggcatggca gacttgaaaa agtcatgttg
      961 tttcatatga tctgggtatc tcgcaaaaca ttgaacacca taaccgaaag tagtgactaa
     1021 ggttggccat ggaactggca atttaccagt agtacaaata aattttaagg tcaatttacc
     1081 gtaagtagca tcaccttcac cttcaccgga gacagaaaat ttgtgaccat taacatcacc
     1141 atctaattca accaaaattg ggacaacacc agtgaataat tcttcacctt tagaggatcc
     1201 catagaaccg ccgccaccgg agccaccgcc accagaacca ccaccacctg gagtgatgtc
     1261 accatacaaa cccaaaatca atggcaacat ccaatctgga gtaactcttt gatgagaaac
     1321 aacttgagct tgataaaatt taataacttc accagattca tcttgaatat gagcagctgg
     1381 agttggacca ccattaccca ttttatcgat ttatattgaa ttttcaaaaa ttcttacttt
     1441 tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcaata ccaccatata
     1501 catatccata tctaatctta cttatatgtt gtggaaatgt aaagagcccc attatcttag
     1561 cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt
     1621 gaagtacgga ttagaagccg ccgagcgggc gacagccctc cgacggaaga ctctcctccg
     1681 tgcgtcctgg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     1741 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     1801 gtaacctggc cccacaaacc ttcaaatcaa cgaatcaaat taacaaccat aggataataa
     1861 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     1921 tctattaaca gatatataaa tgcaaaagct gcataaccac tttaactaat actttcaaca
     1981 ttttcggttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa aaattgttaa
     2041 tatacctcta tactttaacg tcaaggagaa aaaactatag ctagcaaaat ggcttcaggt
     2101 gtaagtaaat gcgaaaccaa atgcacaaag gcttgcccta gaccagctcc agttcctaag
     2161 ttattgatta aaggtggtat ggaagttttg gatttggtta caggtccaga ttctgttact
     2221 gaaatcgaag catttttgaa cccaagaatg ggtcaaccac caacaccaga atctttaact
     2281 gaaggtggtc aatattacgg ttggtcaaga ggtattaatt tggctacatc tgatactgaa
     2341 gattcaccag gtaataatac attaccaact tggtctatgg caaaattgca attaccaatg
     2401 ttgaacgaag atttgacttg tgatactttg caaatgtggg aagcagtttc agttaaaaca
     2461 gaagttgttg gttctggttc tttgttggat gttcatggtt ttaataagcc aacagatact
     2521 gttaacacaa agggtatttc tactccagtt gaaggttcac aatatcatgt ttttgctgtt
     2581 ggtggtgaac cattggattt gcaaggttta gttacagatg caagaactaa gtacaaggaa
     2641 gaaggtgttg ttactattaa aacaatcact aagaaagata tggttaataa ggatcaagtt
     2701 ttgaacccaa tttcaaaggc taaattggat aaggatggca tgtacccagt tgaaatttgg
     2761 catccagatc cagctaaaaa tgaaaacaca agatacttcg gtaattacac tggtggtact
     2821 acaactccac cagttttgca attcactaac actttgacaa ctgttttgtt agatgaaaat
     2881 ggtgttggtc cattatgtaa aggtgaaggt ttgtacttat cttgcgttga tattatgggt
     2941 tggagagtta caagaaacta cgatgttcat cattggagag gtttgccaag atacttcaag
     3001 attactttaa gaaaaagatg ggttaaaaat ccatacccat aagcggccgc actagtagga
     3061 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     3121 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     3181 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat
     3241 ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3361 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5461 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5521 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5581 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5641 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5701 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5761 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5821 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5881 acgttgtaaa acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa
     5941 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     6001 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc
     6061 aggaaacgaa gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca
     6121 agtccggttg catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct
     6181 tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac
     6241 atttgccttt tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc
     6301 gttccattga aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc
     6361 gccgatatcg gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa
     6421 tggtctgata tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa
     6481 caaggtgcgc aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct
     6541 tccaagggtt ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt
     6601 gataaagatt tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg
     6661 tttgattggc taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt
     6721 cagcagtaca gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc
     6781 agaggacttt t
//
