LOCUS       sequence-330315-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative plasmid for promoter-reporter 
            4xUAS-SSA1p-365:yEGFP3-CYC1t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  AUTHORS   Romero-Suarez D, Wulff T, Rong Y, Jakociu Nas T, Yuzawa S, Keasling 
            JD, Jensen MK
  TITLE     A Reporter System for Cytosolic Protein Aggregates in Yeast.
  JOURNAL   ACS Synth Biol. 2021 Feb 12. doi: 10.1021/acssynbio.0c00476.
  PUBMED    33577304
REFERENCE   2  (bases 1 to 5879)
  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: "ACS Synth 
            Biol. 2021 Feb 12. doi: 10.1021/acssynbio.0c00476."
COMMENT     Sequence Label: pYR69 (X-4:4xUAS-SSA1p-365:yEGFP3-CYC1t)
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1005..1721
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1731..1920
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1937..1956)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1938..1956)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1966..1982)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1975..1997)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2531..2564)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(3034..3050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3149..3166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3320..3908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3400..3419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4030..4821,4822..4890))
                     /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     4653..4672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4891..4982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5024..5043
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5049..5068
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5275..5308
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 tttccagaac attctagaaa gaaagcacac ggaacgttta gaagctgtca tttgcgtttt
       61 ttctccagat tttagttgag aaagtaatta aattattctt ctttttccag aacgttccat
      121 cggcggcaaa agggagagaa agaacccaaa aagaaggggg gccatttaga ttagctgatc
      181 gtttcgagga cttcaaggta ctgcatgttt tccagaacat tctagaaaga aagcacacgg
      241 aacgtttaga agctgtcatt tgcgtttttt ctccagattt tagttgagaa agtaattaaa
      301 ttattcttct ttttccagaa cgttccatcg gcggcaaaag ggagagaaag aacccaaaaa
      361 gaaggggggc catttagatt agctgatcgt ttcgaggact tcaaggtacc tgcacttttc
      421 cagaacattc tagaaagaaa gcacacggaa cgtttagaag ctgtcatttg cgttttttct
      481 ccagatttta gttgagaaag taattaaatt attcttcttt ttccagaacg ttccatcggc
      541 ggcaaaaggg agagaaagaa cccaaaaaga aggggggcca tttagattag ctgatcgttt
      601 cgaggacttc aaggtacgtg cgattttcca gaacattcta gaaagaaagc acacggaacg
      661 tttagaagct gtcatttgcg ttttttctcc agattttagt tgagaaagta attaaattat
      721 tcttcttttt ccagaacgtt ccatcggcgg caaaagggag agaaagaacc caaaaagaag
      781 gggggccatt tagattagct gatcgtttcg aggacttcaa ggttatataa ggggtggatt
      841 gatgtatctt cgagaaggga ttgagttgta gtttcgtttc ccaattctta cttaagttgt
      901 tttattttct ctatttgtaa gataagcaca tcaaaagaaa agtaatcaag tattacaaga
      961 aacaaaaatt caagtaaata acagataata cttgtcataa aacaatgtct aaaggtgaag
     1021 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
     1081 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
     1141 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt
     1201 atggtgttca atgttttgcg agatacccag atcatatgaa acaacatgac tttttcaagt
     1261 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     1321 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     1381 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     1441 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     1501 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     1561 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg
     1621 ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     1681 ctgctggtat tacccatggt atggatgaat tgtacaaatg aacacgcgat atccgctcta
     1741 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     1801 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     1861 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     1921 atcgcgtccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     1981 acaacgtcgt gactgggaaa accctggcgt tacccctgca ggactagtgc tgaggcatta
     2041 atacatggga agattcgctt tttttttttg aattacaata gtatgtctga tgtctgcaag
     2101 aagtaacagg cgtgtgcaca agaatacgtg tgtgtgcgta agcgtatgca ctggtggcat
     2161 aacttatcta agaagtatat atcactgaca tagaaatgta gatatacagg tatttttctc
     2221 gataatcgat aaaaatctcg tcgcgctgaa ccaaacttgg tggttacgga gagtttttct
     2281 ctcatcatta ctgtctttcg cattgatttc ccctttgacc gataaaatcc cttggattca
     2341 taagattaaa caaagaggtg atcaaagaga accctgtgaa agtttatgtt tataaccggg
     2401 cataaagtga actagacact ttcaagaagc caaccaaagc atgagtaacg aagcttacca
     2461 gcatgatcat accgtaaatc ctcaccagat taagacctca gcgcggccgc aaatttaaat
     2521 aaaatgaagt gaagttccta tactttctag agaataggaa cttctatagt gagtcgaata
     2581 agggcgacac aaaatttatt ctaaatgcat aataaatact gataacatct tatagtttgt
     2641 attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt
     2701 tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac
     2761 aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca
     2821 acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata
     2881 tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc
     2941 ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg
     3001 ttatcagaac cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa
     3061 gagtttgtag aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg
     3121 gcagttccct actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3721 agttcttgaa gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg
     3781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc
     3961 gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta
     4021 atgctctgct taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4081 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4141 atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc
     4201 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4261 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4321 tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4381 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4441 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4501 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4561 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4621 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4681 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4741 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4801 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4861 aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt attgaagcat
     4921 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4981 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
     5041 acctgaatat ggctcataac accccttgtt tgcctggcgg cagtagcgcg gtggtcccac
     5101 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggactc
     5161 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     5221 tgggcctttc gcccgggcta attatggggt gtcgccctta ttcgactcta tagtgaagtt
     5281 cctattctct agaaagtata ggaacttctg aagtggggat ttaaatgcgg ccgcgctgag
     5341 ggtttaatcc caaagctaag agtcccattt tattcttcta tatgtatatt ttcgatactc
     5401 taaaccaccc tacaatgtag ccctatacta aatctgctca attttcagct tctacaagtg
     5461 actcgaggcc acgtggaaag atccaactac tccagcacaa cgattcaata taatcgattg
     5521 ctccactcat aagaggcaag aacaagcttc aacttttggt aagccgccgt ttataaacag
     5581 ggaagatgtc ctttgtcaag ggaggcacag agcatggcca atttggcaaa ttgcaggttt
     5641 ttctgagtga aaaatgaaaa agcattgtag tagagtcggc tcactgaaaa accggggagg
     5701 acgaaaaggt ttccagccac agttgtagtc acgtgcgcgc catgctgact aatggcagcc
     5761 gtcgttgggc agaagagaat tagtatggta caggatacgc taattgcgct ccaactacca
     5821 aggttgttga gggaacactg gggcaatagg ctgtcgccat tcaagagcag agacacttt
//
