LOCUS       sequence-329833-        6992 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast integrative plasmid for expressing wild-type Murine 
            polyomavirus VP1 (GAL1 promoter) and destabilised GFP tagged with 
            VP2C (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 6992)
  AUTHORS   Cheah LC, Stark T, Adamson LSR, Abidin RS, Lau YH, Sainsbury F, 
            Vickers CE
  TITLE     Artificial Self-assembling Nanocompartment for Organizing Metabolic 
            Pathways in Yeast.
  JOURNAL   ACS Synth Biol. 2021 Dec 17;10(12):3251-3263. doi: 
            10.1021/acssynbio.1c00045. Epub 2021 Sep 30.
  PUBMED    34591448
REFERENCE   2  (bases 1 to 6992)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.1c00045"; journalName: "ACS Synth Biol"; date: 
            "2021-12-17- 17"; volume: "10"; issue: "12"; pages: "3251-3263"
COMMENT     Sequence Label: wtVP1 + VP2C-GFPdeg
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(235..251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(235..251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(248..270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    259..275
                     /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(283..289,290..307,308..313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    328..349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(466..483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(637..1225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(717..736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1396..2187,2188..2256))
                     /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     2019..2038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2257..2361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2429..2447
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2485..2507)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2607..2626
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2820..2842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2834..2851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2835..2851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4224..4934)
                     /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        5367..5808
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5367..5484
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5779..5802
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
       61 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
      121 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
      181 cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat catggtcata
      241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     2461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2821 ccagtcacga cgttgtaaaa cgacggccag tgaattcatt taaatttttt tgattcggta
     2881 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2941 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     3001 aaaacctgca ggaaacgaag ataaaacatg tccacaaaat catataccag tagagctgag
     3061 actcatgcaa gtccggttgc atcgaaactt ttacgtttaa tggatgaaaa gaagaccaat
     3121 ttgtgtgctt ctcttgacgt tcgttcgact gatgagctat tgaaacttgt tgaaacgttg
     3181 ggtccataca tttgcctttt gaaaacacac gttgatatct tggatgattt cagttatgag
     3241 ggtactgtcg ttccattgaa agcattggca gagaaataca agttcttgat atttgaggac
     3301 agaaaattcg ccgatatcgg taacacagtc aaattacaat atacatcggg cgtttaccgt
     3361 atcgcagaat ggtctgatat caccaacgcc cacggggtta ctggtgctgg tattgttgct
     3421 ggcttgaaac aaggtgcgca agaggtcacc aaagaaccaa ggggattatt gatgcttgct
     3481 gaattgtctt ccaagggttc tctagcacac ggtgaatata ctaagggtac cgttgatatt
     3541 gcaaagagtg ataaagattt cgttattggg ttcattgctc agaacgatat gggaggaaga
     3601 gaagaagggt ttgattggct aatcatgacc ccaggtgtag gtttagacga caaaggcgat
     3661 gcattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc agatatcatc
     3721 attgttggca gaggactttt cgccaagggt agagatccta aggttgaagg tgaaagatac
     3781 agaaatgctg gatgggaagc gtaccaaaag agaatcagcg ctccccatta attatacagg
     3841 aaacttaata gaacaaatca catatttaat ctaatagcca cctgcattgg cacggtgcaa
     3901 cactacttca acttcatctt acaaaaagat cacgtgatct gttgtattgg gatctctaga
     3961 gtaacaatat catgagacct tttatagaag tggcgccaaa actaaatgta tttgaaaata
     4021 caaaaaacgc acaggtttct ttatatatgt atatatatat atatatatat aaaactgtac
     4081 atatactgtt taaattaatc tatttaaaca ttaattcttg cagatgcaca agcagctgga
     4141 tgcctatcca tacctgattc ttgtgcacat gacattggca aagttccatc atcttgctct
     4201 tcaacttcag gaggaaaaga tcttttgtac aattcatcca taccatgggt aataccagca
     4261 gcagtaacaa attctaacaa gaccatgtgg tctctctttt cgtttggatc tttggataag
     4321 gcagattgag tggataagta atggttgtct ggtaacaaga ctggaccatc accaattgga
     4381 gtattttgtt gataatggtc agctaattga acagaaccat cttcaatgtt gtgtctaatt
     4441 ttgaagttaa ctttgatacc attcttttgt ttgtcagcca tgatgtaaac attgtgagag
     4501 ttatagttgt attccaattt gtgacctaaa atgttaccat cttctttaaa atcaatacct
     4561 tttaattcga ttctattaac taaggtatca ccttcaaact tgacttcagc tctggtcttg
     4621 tagttaccgt catctttgaa aaaaatagtt ctttcttgaa cataaccttc tggcatggca
     4681 gacttgaaaa agtcatgttg tttcatatga tctgggtatc tcgcaaaaca ttgaacacca
     4741 taaccgaaag tagtgactaa ggttggccat ggaactggca atttaccagt agtacaaata
     4801 aattttaagg tcaatttacc gtaagtagca tcaccttcac cttcaccgga gacagaaaat
     4861 ttgtgaccat taacatcacc atctaattca accaaaattg ggacaacacc agtgaataat
     4921 tcttcacctt tagaggatcc catagaaccg ccgccaccgg agccaccgcc accagaacca
     4981 ccaccacctg gagtgatgtc accatacaaa cccaaaatca atggcaacat ccaatctgga
     5041 gtaactcttt gatgagaaac aacttgagct tgataaaatt taataacttc accagattca
     5101 tcttgaatat gagcagctgg agttggacca ccattaccca ttttatcgat ttatattgaa
     5161 ttttcaaaaa ttcttacttt tttttttgga tggacgcaaa gaagtttaat aatcatatta
     5221 catggcaata ccaccatata catatccata tctaatctta cttatatgtt gtggaaatgt
     5281 aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct
     5341 taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggc gacagccctc
     5401 cgacggaaga ctctcctccg tgcgtcctgg tcttcaccgg tcgcgttcct gaaacgcaga
     5461 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
     5521 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatcaa cgaatcaaat
     5581 taacaaccat aggataataa tgcgattagt tttttagcct tatttctggg gtaattaatc
     5641 agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaagct gcataaccac
     5701 tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat gtcataaaag
     5761 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactatag
     5821 ctagcaaaat ggctccaaag agaaagtcag gtgtaagtaa atgcgaaacc aaatgcacaa
     5881 aggcttgccc tagaccagct ccagttccta agttattgat taaaggtggt atggaagttt
     5941 tggatttggt tacaggtcca gattctgtta ctgaaatcga agcatttttg aacccaagaa
     6001 tgggtcaacc accaacacca gaatctttaa ctgaaggtgg tcaatattac ggttggtcaa
     6061 gaggtattaa tttggctaca tctgatactg aagattcacc aggtaataat acattaccaa
     6121 cttggtctat ggcaaaattg caattaccaa tgttgaacga agatttgact tgtgatactt
     6181 tgcaaatgtg ggaagcagtt tcagttaaaa cagaagttgt tggttctggt tctttgttgg
     6241 atgttcatgg ttttaataag ccaacagata ctgttaacac aaagggtatt tctactccag
     6301 ttgaaggttc acaatatcat gtttttgctg ttggtggtga accattggat ttgcaaggtt
     6361 tagttacaga tgcaagaact aagtacaagg aagaaggtgt tgttactatt aaaacaatca
     6421 ctaagaaaga tatggttaat aaggatcaag ttttgaaccc aatttcaaag gctaaattgg
     6481 ataaggatgg catgtaccca gttgaaattt ggcatccaga tccagctaaa aatgaaaaca
     6541 caagatactt cggtaattac actggtggta ctacaactcc accagttttg caattcacta
     6601 acactttgac aactgttttg ttagatgaaa atggtgttgg tccattatgt aaaggtgaag
     6661 gtttgtactt atcttgcgtt gatattatgg gttggagagt tacaagaaac tacgatgttc
     6721 atcattggag aggtttgcca agatacttca agattacttt aagaaaaaga tgggttaaaa
     6781 atccataccc aatggcttct ttgatttctt ctttgtttaa taatatgtta ccacaagttc
     6841 aaggtcaacc aatggaaggt gaaaacacac aagttgaaga agttagagtt tacgatggta
     6901 ctgaaccagt tccaggtgac cctgacatga caagatatgt agatagattc ggtaaaacta
     6961 aaactgtatt ccctggtaat taagcggccg ca
//
