LOCUS       sequence-325281-        5882 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 5882)
  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 5882)
  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..5882
                     /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(1938..1957)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1939..1957)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1967..1983)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1976..1998)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2532..2565)
                     /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(3035..3051)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3150..3167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3321..3909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3401..3420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4033..4824,4825..4893))
                     /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     4656..4675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4894..4985)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5027..5046
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5052..5071
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5278..5311
                     /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 atcgcgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     1981 tacaacgtcg tgactgggaa aaccctggcg ttacccctgc aggactagtg ctgaggcatt
     2041 aatacatggg aagattcgct tttttttttt gaattacaat agtatgtctg atgtctgcaa
     2101 gaagtaacag gcgtgtgcac aagaatacgt gtgtgtgcgt aagcgtatgc actggtggca
     2161 taacttatct aagaagtata tatcactgac atagaaatgt agatatacag gtatttttct
     2221 cgataatcga taaaaatctc gtcgcgctga accaaacttg gtggttacgg agagtttttc
     2281 tctcatcatt actgtctttc gcattgattt cccctttgac cgataaaatc ccttggattc
     2341 ataagattaa acaaagaggt gatcaaagag aaccctgtga aagtttatgt ttataaccgg
     2401 gcataaagtg aactagacac tttcaagaag ccaaccaaag catgagtaac gaagcttacc
     2461 agcatgatca taccgtaaat cctcaccaga ttaagacctc agcgcggccg caaatttaaa
     2521 taaaatgaag tgaagttcct atactttcta gagaatagga acttctatag tgagtcgaat
     2581 aagggcgaca caaaatttat tctaaatgca taataaatac tgataacatc ttatagtttg
     2641 tattatattt tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct
     2701 ttattatttt cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata
     2761 caaaaaatca taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc
     2821 aacgtatctt atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat
     2881 atatcaagca aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc
     2941 cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc
     3001 gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa
     3061 agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct
     3121 ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3721 gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc
     3781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
     3961 cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt
     4021 aatgctctgc ttttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4141 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
     4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4321 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4861 aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat attattgaag
     4921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4981 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt
     5041 tatacctgaa tatggctcat aacacccctt gtttgcctgg cggcagtagc gcggtggtcc
     5101 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtgggga
     5161 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     5221 gactgggcct ttcgcccggg ctaattatgg ggtgtcgccc ttattcgact ctatagtgaa
     5281 gttcctattc tctagaaagt ataggaactt ctgaagtggg gatttaaatg cggccgcgct
     5341 gagggtttaa tcccaaagct aagagtccca ttttattctt ctatatgtat attttcgata
     5401 ctctaaacca ccctacaatg tagccctata ctaaatctgc tcaattttca gcttctacaa
     5461 gtgactcgag gccacgtgga aagatccaac tactccagca caacgattca atataatcga
     5521 ttgctccact cataagaggc aagaacaagc ttcaactttt ggtaagccgc cgtttataaa
     5581 cagggaagat gtcctttgtc aagggaggca cagagcatgg ccaatttggc aaattgcagg
     5641 tttttctgag tgaaaaatga aaaagcattg tagtagagtc ggctcactga aaaaccgggg
     5701 aggacgaaaa ggtttccagc cacagttgta gtcacgtgcg cgccatgctg actaatggca
     5761 gccgtcgttg ggcagaagag aattagtatg gtacaggata cgctaattgc gctccaacta
     5821 ccaaggttgt tgagggaaca ctggggcaat aggctgtcgc cattcaagag cagagacact
     5881 tt
//
