LOCUS       sequence-373864-        6672 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 6672)
  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 6672)
  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: pYES2/25Q
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(198..445)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     411..429
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(460..1173,1174..1176))
                     /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             complement(join(463..1173,1174..1176))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(505..526)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     854..873
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1113..1134
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1324..1347)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1372..1391)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1373..1391)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1423..1864)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1429..1452)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1747..1864
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      1886..2399
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1973..1992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2183..2204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2410..3287
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3888..4103
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4104..4907
                     /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"
     CDS             join(5003..5071,5072..5863)
                     /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     complement(5221..5240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6034..6622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6523..6542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
       61 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      121 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      181 atgcagggcc gcagcttgca aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag
      241 gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga
      301 aatataaata acgttcttaa tactaacata actataaaaa aataaatagg gacctagact
      361 tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaatgt
      421 aagcgtgaca taactaatta catgatgcgg ccctctagat tacttgtaca gctcgtccat
      481 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      541 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
      601 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      661 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
      721 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
      781 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      841 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      901 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      961 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1021 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1081 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1141 caccaccccg gtgaacagct cctcgccctt gctcaccagg gatcccccgg gctgcagttg
     1201 ttgctgttgc tgctgttgtt gctgttgctg ctgttgttgc tgttgctgct gttgttgctg
     1261 ttgctgctgt tggaagcttt tgagggactc gaaggccttc atcagctttt ccagggtcgc
     1321 cagcttgtca tcgtcgtcct tgtagtccat ggcggtacca agcttaatat tccctatagt
     1381 gagtcgtatt acagctgcta gtagtccgat ccggggtttt ttctccttga cgttaaagta
     1441 tagaggtata ttaacaattt tttgttgata cttttattac atttgaataa gaagtaatac
     1501 aaaccgaaaa tgttgaaagt attagttaaa gtggttatgc agtttttgca tttatatatc
     1561 tgttaataga tcaaaaatca tcgcttcgct gattaattac cccagaaata aggctaaaaa
     1621 actaatcgca ttatcatcct atggttgtta atttgattcg ttcatttgaa ggtttgtggg
     1681 gccaggttac tgccaatttt tcctcttcat aaccataaaa gctagtattg tagaatcttt
     1741 attgttcgga gcagtgcggc gcgaggcaca tctgcgtttc aggaacgcga ccggtgaaga
     1801 cgaggacgca cggaggagag tcttccttcg gagggctgtc acccgctcgg cggcttctaa
     1861 tccgtggact agtggatcga tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1921 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1981 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2041 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2101 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2161 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2221 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2281 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2341 taaatatttg cttatacaat cttcctgttt ttggggcttt tctgattatc aaccggggtg
     2401 gagcttccca ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata
     2461 tatctctgtg ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca
     2521 tctaaactcg acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa
     2581 attgtagtaa gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc
     2641 tatacgtagt atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag
     2701 aatcatcaac gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat
     2761 ataatcgggg atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa
     2821 acgcgggaag tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt
     2881 cttctaacct taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc
     2941 acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc
     3001 atttttgtag aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt
     3061 gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc
     3121 gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc
     3181 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     3241 ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttaac aaagatatgc
     3301 tattgaagtg caagatggaa acgcagaaaa tgaaccgggg atgcgacgtg caagattacc
     3361 tatgcaatag atgcaatagt ttctccagga accgaaatac atacattgtc ttccgtaaag
     3421 cgctagacta tatattatta tacaggttca aatatactat ctgtttcagg gaaaactccc
     3481 aggttcggat gttcaaaatt caatgatggg taacaagtac gatcgtaaat ctgtaaaaca
     3541 gtttgtcgga tattaggctg tatctcctca aagcgtattc gaatatcatt gagaagctgc
     3601 agcgtcacat cggataataa tgatggcagc cattgtagaa gtgccttttg catttctagt
     3661 ctctttctcg gtctagctag ttttactaca tcgcgaagat agaatcttag atcacactgc
     3721 ctttgctgag ctggatcaat agagtaacaa aagagtggta aggcctcgtt aaaggacaag
     3781 gacctgagcg gaagtgtatc gtacagtaga cggagtatac taggtatagt ctatagtccg
     3841 tggaattaat tctcatgttt gacagcttat catcgataag ctagcttttc aattcatcat
     3901 ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct
     3961 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     4021 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     4081 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     4141 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     4201 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     4261 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     4321 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
     4381 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     4441 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     4501 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     4561 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     4621 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     4681 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
     4741 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
     4801 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     4861 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     4921 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     4981 tacccattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5401 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5521 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5701 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5821 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5881 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5941 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6661 ttgctggcct tt
//
