LOCUS       sequence-360112-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing Murine polyomavirus 
            deltaVP1 (GAL1 promoter) and fusion protein VP2C-GFP-mRuby3 (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 7609)
  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 7609)
  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 mRuby3 VLP
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(310..326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(310..326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(323..345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    334..350
                     /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(358..364,365..382,383..388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    403..424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(541..558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(712..1300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(792..811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1471..2262,2263..2331))
                     /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     2094..2113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2332..2436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2504..2522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2560..2582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2682..2701
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2895..2917
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2909..2926
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2910..2926
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4184..4894)
                     /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(4928..5638)
                     /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        6071..6512
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6071..6188
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6483..6506
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 gaccctgaca tgacaagata tgtagataga ttcggtaaaa ctaaaactgt attccctggt
       61 aattaagcgg ccgcactagt aggactattt gcaaagggaa gggatgctaa ggtagagggt
      121 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
      181 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
      241 ttatatcagt tattacccgg gaatctcggt cgtaatgatt taaatgcatg caagcttggc
      301 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2461 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2521 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2581 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2641 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     2701 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2761 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     2821 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     2881 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcatttaaat
     2941 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     3001 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     3061 caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac aaaatcatat
     3121 accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg tttaatggat
     3181 gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga gctattgaaa
     3241 cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga tatcttggat
     3301 gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa atacaagttc
     3361 ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt acaatataca
     3421 tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg ggttactggt
     3481 gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga accaagggga
     3541 ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga atatactaag
     3601 ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat tgctcagaac
     3661 gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg tgtaggttta
     3721 gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt tgtaagtggt
     3781 ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga tcctaaggtt
     3841 gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat cagcgctccc
     3901 cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat agccacctgc
     3961 attggcacgg tgcaacacta cttcaacttc atcttacaaa aagatcacgt gatctgttgt
     4021 attgggatct ctagagtaac aatatcatga gaccttttat agaagtggcg ccaaaactaa
     4081 atgtatttga aaatacaaaa aacgcacagg tttctttata tatgtatata tatatatata
     4141 tatatataaa actgtacata tactgtttaa attaatctat ttacttgtac agctcgtcca
     4201 tgccaccacc aaggttgctg tatttggcaa ctgccacttc gcgttgcact acgtaggttt
     4261 cattgtcact ctcctcgatc ctttccaggc ggtgatcaac ggcatggaca ccgggcatct
     4321 tgatgttccc gacggtcttt tttgacctgt aagttgtcac gaagttgcag tgcagatggc
     4381 caccaccatc aactttcagt gcgatgtcag tgtatcctct cagaccacca tctgctggat
     4441 acatcatctc tgtattaggc tcccaaccct tggtcttctt ctgcatcacg ggaccattgg
     4501 agggaaagtt tacccctctg accttgacgt tgtagacgag ctcgccatcc tcaaggctgg
     4561 tgtcctgcgt gacggtgacg actccaccat cttcgtatct cgtaactctt tcccaagtaa
     4621 aaccctcagg aaaggactgt ttaaagaaat cagggatgtc ggccgggtac ttgataaaag
     4681 tacggctgcc atacatgaac gacgtggcaa gaatgtcaaa ggcaaatggc aggggtcctc
     4741 cctcgatgac tttgatcctc atggtttgca ctccctcgta cggtctgcct tctccttcac
     4801 ctgtgcattt gaattggtgg ccgttgaccg aaccttccat gaccaccttc atacgcatat
     4861 tttccttgat cagctcttcg cccttagaca ccatagcacc accagtacct cctccaccag
     4921 cagatctttt gtacaattca tccataccat gggtaatacc agcagcagta acaaattcta
     4981 acaagaccat gtggtctctc ttttcgtttg gatctttgga taaggcagat tgagtggata
     5041 agtaatggtt gtctggtaac aagactggac catcaccaat tggagtattt tgttgataat
     5101 ggtcagctaa ttgaacagaa ccatcttcaa tgttgtgtct aattttgaag ttaactttga
     5161 taccattctt ttgtttgtca gccatgatgt aaacattgtg agagttatag ttgtattcca
     5221 atttgtgacc taaaatgtta ccatcttctt taaaatcaat accttttaat tcgattctat
     5281 taactaaggt atcaccttca aacttgactt cagctctggt cttgtagtta ccgtcatctt
     5341 tgaaaaaaat agttctttct tgaacataac cttctggcat ggcagacttg aaaaagtcat
     5401 gttgtttcat atgatctggg tatctcgcaa aacattgaac accataaccg aaagtagtga
     5461 ctaaggttgg ccatggaact ggcaatttac cagtagtaca aataaatttt aaggtcaatt
     5521 taccgtaagt agcatcacct tcaccttcac cggagacaga aaatttgtga ccattaacat
     5581 caccatctaa ttcaaccaaa attgggacaa caccagtgaa taattcttca cctttagagg
     5641 atcccataga accgccgcca ccggagccac cgccaccaga accaccacca cctggagtga
     5701 tgtcaccata caaacccaaa atcaatggca acatccaatc tggagtaact ctttgatgag
     5761 aaacaacttg agcttgataa aatttaataa cttcaccaga ttcatcttga atatgagcag
     5821 ctggagttgg accaccatta cccattttat cgatttatat tgaattttca aaaattctta
     5881 cttttttttt tggatggacg caaagaagtt taataatcat attacatggc aataccacca
     5941 tatacatatc catatctaat cttacttata tgttgtggaa atgtaaagag ccccattatc
     6001 ttagcctaaa aaaaccttct ctttggaact ttcagtaata cgcttaactg ctcattgcta
     6061 tattgaagta cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc
     6121 tccgtgcgtc ctggtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca
     6181 ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt
     6241 ggcagtaacc tggccccaca aaccttcaaa tcaacgaatc aaattaacaa ccataggata
     6301 ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt
     6361 ttgatctatt aacagatata taaatgcaaa agctgcataa ccactttaac taatactttc
     6421 aacattttcg gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg
     6481 ttaatatacc tctatacttt aacgtcaagg agaaaaaact atagctagca aaatggcttc
     6541 aggtgtaagt aaatgcgaaa ccaaatgcac aaaggcttgc cctagaccag ctccagttcc
     6601 taagttattg attaaaggtg gtatggaagt tttggatttg gttacaggtc cagattctgt
     6661 tactgaaatc gaagcatttt tgaacccaag aatgggtcaa ccaccaacac cagaatcttt
     6721 aactgaaggt ggtcaatatt acggttggtc aagaggtatt aatttggcta catctgatac
     6781 tgaagattca ccaggtaata atacattacc aacttggtct atggcaaaat tgcaattacc
     6841 aatgttgaac gaagatttga cttgtgatac tttgcaaatg tgggaagcag tttcagttaa
     6901 aacagaagtt gttggttctg gttctttgtt ggatgttcat ggttttaata agccaacaga
     6961 tactgttaac acaaagggta tttctactcc agttgaaggt tcacaatatc atgtttttgc
     7021 tgttggtggt gaaccattgg atttgcaagg tttagttaca gatgcaagaa ctaagtacaa
     7081 ggaagaaggt gttgttacta ttaaaacaat cactaagaaa gatatggtta ataaggatca
     7141 agttttgaac ccaatttcaa aggctaaatt ggataaggat ggcatgtacc cagttgaaat
     7201 ttggcatcca gatccagcta aaaatgaaaa cacaagatac ttcggtaatt acactggtgg
     7261 tactacaact ccaccagttt tgcaattcac taacactttg acaactgttt tgttagatga
     7321 aaatggtgtt ggtccattat gtaaaggtga aggtttgtac ttatcttgcg ttgatattat
     7381 gggttggaga gttacaagaa actacgatgt tcatcattgg agaggtttgc caagatactt
     7441 caagattact ttaagaaaaa gatgggttaa aaatccatac ccaatggctt ctttgatttc
     7501 ttctttgttt aataatatgt taccacaagt tcaaggtcaa ccaatggaag gtgaaaacac
     7561 acaagttgaa gaagttagag tttacgatgg tactgaacca gttccaggt
//
