LOCUS       sequence-243790-        6150 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 6150)
  AUTHORS   Duennwald ML, Jagadish S, Muchowski PJ, Lindquist S
  TITLE     Flanking sequences profoundly alter polyglutamine toxicity in yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Jul 18. 103(29):11045-50.
  PUBMED    16832050
REFERENCE   2  (bases 1 to 6150)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Jul 18. 103(29):11045-50."
COMMENT     Sequence Label: GAL FLAG25Q+ProGFPp303
FEATURES             Location/Qualifiers
     source          1..6150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             469..492
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             778..1488
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(805..833)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1428..1448
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1498..1745
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1514..1532)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1762..1778)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1763..1779)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1807..1825)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1846..1862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1846..1862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1859..1881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1870..1886
                     /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(1894..1900,1901..1918,1919..1924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1939..1960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2077..2094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3007..3798,3799..3867))
                     /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     3630..3649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3868..3972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4040..4058
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4096..4118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4218..4237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4384..4571
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             4572..5234
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(5494..5949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5631..5652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5843..5862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6075..6097
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6089..6106
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6090..6106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6113..6132
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6113..6131
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggattcta gagccatgga ctacaaggac
      481 gacgatgaca agctggcgac cctggaaaag ctgatgaagg ccttcgagtc cctcaaaagc
      541 ttccaacagc agcaacagca acaacagcag caacagcaac aacagcagca acagcaacaa
      601 cagcagcaac agcaacaacc gccaccacct ccccctccac ccccacctcc tcaacttcct
      661 caacctcctc cacaggcaca gcctctgctg cctcagccac aacctcctcc acctccacct
      721 ccacctcctc caggcccagc tgtggctgag gagcctctgc accgacctgg atccctgagt
      781 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
      841 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
      901 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
      961 actttgactt ggggtgttca atgcttttca agatacccag atcatatgaa acggcatgac
     1021 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     1081 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     1141 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     1201 tacaactata tttcacacaa tgtatacatc actgcagaca aacaaaagaa tggaatcaaa
     1261 gctcatttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     1321 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     1381 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     1441 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata gctcgagtca
     1501 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     1561 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     1621 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     1681 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     1741 tttgcggccg atcaagctta tcgataccgt cgacctcgag ggggggcccg gtaccagctt
     1801 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     1861 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1921 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1981 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4021 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4081 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4141 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4201 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaattccc
     4261 gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa cacggcatta
     4321 gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta ctcttggcct
     4381 cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt tttttttcca
     4441 cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat gaattataca
     4501 ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag gcaagataaa
     4561 cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg tattacaaat gaaaccaaga
     4621 ttcagattgc gatctcttta aagggtggtc ccctagcgat agagcactcg atcttcccag
     4681 aaaaagaggc agaagcagta gcagaacagg ccacacaatc gcaagtgatt aacgtccaca
     4741 caggtatagg gtttctggac catatgatac atgctctggc caagcattcc ggctggtcgc
     4801 taatcgttga gtgcattggt gacttacaca tagacgacca tcacaccact gaagactgcg
     4861 ggattgctct cggtcaagct tttaaagagg ccctaggggc cgtgcgtgga gtaaaaaggt
     4921 ttggatcagg atttgcgcct ttggatgagg cactttccag agcggtggta gatctttcga
     4981 acaggccgta cgcagttgtc gaacttggtt tgcaaaggga gaaagtagga gatctctctt
     5041 gcgagatgat cccgcatttt cttgaaagct ttgcagaggc tagcagaatt accctccacg
     5101 ttgattgtct gcgaggcaag aatgatcatc accgtagtga gagtgcgttc aaggctcttg
     5161 cggttgccat aagagaagcc acctcgccca atggtaccaa cgatgttccc tccaccaaag
     5221 gtgttcttat gtagtgacac cgattattta aagctgcagc atacgatata tatacatgtg
     5281 tatatatgta tacctatgaa tgtcagtaag tatgtatacg aacagtatga tactgaagat
     5341 gacaaggtaa tgcatcattc tatacgtgtc attctgaacg aggcgcgctt tccttttttc
     5401 tttttgcttt ttcttttttt ttctcttgaa ctcgacggat ctatgcggtg tgaaataccg
     5461 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     5521 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5581 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5641 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5701 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5761 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5821 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5881 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5941 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6001 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6061 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     6121 ctcactatag ggcgaattgg agctctagta
//
