LOCUS       sequence-261011-        6888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  AUTHORS   Meriin AB, Zhang X, He X, Newnam GP, Chernoff YO, Sherman MY
  TITLE     Huntington toxicity in yeast model depends on polyglutamine 
            aggregation mediated by a prion-like protein Rnq1.
  JOURNAL   J Cell Biol 2002 Jun 10;157(6):997-1004. Epub 2002 Jun 10.
  PUBMED    12058016
REFERENCE   2  (bases 1 to 6888)
  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: "J Cell 
            Biol"; date: "2002-06-10"; volume: "157(6)"; pages: "997-1004. Epub 
            2002 Jun 10"
COMMENT     Sequence Label: pYES2/103Q
FEATURES             Location/Qualifiers
     source          1..6888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..487
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    46..163
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     458..481
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     519..538
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        519..537
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             563..586
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(950..952,953..1666)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(950..952,953..1663)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(992..1013)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1253..1272)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1600..1621
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1681..1928
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1697..1715)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2005..2022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2176..2764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2256..2275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2935..3726,3727..3795))
                     /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     3558..3577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3891..4694)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4695..4910)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5511..6388
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(join(6399..6888,1..24))
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6594..6615)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6806..6825
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttaatgcgcc gctacagggc gcgtggggat cgatccacta gtccacggat tagaagccgc
       61 cgagcgggtg acagccctcc gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt
      121 cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac
      181 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct
      241 tcaaatgaac gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt
      301 atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat
      361 gcaaaaactg cataaccact ttaactaata ctttcaacat tttcggtttg tattacttct
      421 tattcaaatg taataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt
      481 caaggagaaa aaaccccgga tcggactact agcagctgta atacgactca ctatagggaa
      541 tattaagctt ggtaccgcca tggactacaa ggacgacgat gacaagctgg cgaccctgga
      601 aaagctgatg aaggccttcg agtccctcaa aagcttccaa cagcagcaac agcaacaaca
      661 gcagcaacag caacaacagc agcaacagca acaacagcag caacagcaac aacagcagca
      721 acagcaacaa cagcagcaac agcaacaaca gcagcaacag caacaacagc agcaacagca
      781 acaacagcag caacagcaac aacagcagca acagcaacaa cagcagcaac agcaacaaca
      841 gcagcaacag caacaacagc agcaacagca acaacagcag caacagcaac aacagcagca
      901 acagcaacaa cagcagcaac agcaacaact gcagcccggg ggatccctgg tgagcaaggg
      961 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1021 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1081 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1141 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1201 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1261 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1321 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1381 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1441 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1501 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1561 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1621 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatcta gagggccgca
     1681 tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg
     1741 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt
     1801 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt
     1861 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt
     1921 taatttgcaa gctgcggccc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3781 atgttgaata ctcatactct tcctttttca atgggtaata actgatataa ttaaattgaa
     3841 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc
     3901 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc
     3961 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta
     4021 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa
     4081 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt
     4141 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta
     4201 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct
     4261 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
     4321 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag
     4381 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa
     4441 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc
     4501 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc
     4561 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
     4621 atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat
     4681 gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt
     4741 aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta
     4801 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt
     4861 caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aagctagctt
     4921 atcgatgata agctgtcaaa catgagaatt aattccacgg actatagact atacctagta
     4981 tactccgtct actgtacgat acacttccgc tcaggtcctt gtcctttaac gaggccttac
     5041 cactcttttg ttactctatt gatccagctc agcaaaggca gtgtgatcta agattctatc
     5101 ttcgcgatgt agtaaaacta gctagaccga gaaagagact agaaatgcaa aaggcacttc
     5161 tacaatggct gccatcatta ttatccgatg tgacgctgca gcttctcaat gatattcgaa
     5221 tacgctttga ggagatacag cctaatatcc gacaaactgt tttacagatt tacgatcgta
     5281 cttgttaccc atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata
     5341 gtatatttga acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta
     5401 tttcggttcc tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc
     5461 cggttcattt tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat
     5521 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     5581 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     5641 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     5701 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     5761 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     5821 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     5881 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     5941 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     6001 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     6061 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     6121 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     6181 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     6241 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     6301 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     6361 caatgtttgt ggaagcggta ttcgcaatgg gaagctccac cccggttgat aatcagaaaa
     6421 gccccaaaaa caggaagatt gtataagcaa atatttaaat tgtaagcgtt aatattttgt
     6481 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6541 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6601 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6661 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6721 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6781 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6841 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgc
//
