LOCUS       sequence-218131-        6599 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 6599)
  AUTHORS   Krobitsch S, Lindquist S
  TITLE     Aggregation of huntingtin in yeast varies with the length of the 
            polyglutamine expansion and the expression of chaperone proteins.
  JOURNAL   Proc Natl Acad Sci U S A 2000 Feb 15;97(4):1589-94.
  PUBMED    10677504
REFERENCE   2  (bases 1 to 6599)
  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"; date: "2000-02-15"; volume: "97(4)"; pages: 
            "1589-94"
COMMENT     Sequence Label: p416 25Q GAL
FEATURES             Location/Qualifiers
     source          1..6599
                     /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"
     primer_bind     457..473
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     457..473
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(507..523)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(508..524)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             join(821..823,824..1537)
                     /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(821..823,824..1534)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(863..884)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1124..1143)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1471..1492
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1553..1800
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1569..1587)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1818..1837)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1819..1837)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1847..1864)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1847..1863)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1856..1878)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2004..2459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2091..2110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2301..2322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2590..3393)
                     /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(3394..3609)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3633..3652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3752..3774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3812..3830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3871..4374
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4411..4515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4516..4584,4585..5376)
                     /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(4734..4753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6289..6306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6423..6444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6459..6464,6465..6482,6483..6489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6497..6513
                     /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)."
     primer_bind     6502..6524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6521..6537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6521..6537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6556..6576
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6558..6576
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 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 gaactagtgg atcccccggg
      481 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgaggtcga ccgccatggc
      541 gaccctggaa aagctgatga aggccttcga gtccctcaaa agcttccaac agcagcaaca
      601 gcaacaacag cagcaacagc aacaacagca gcaacagcaa caacagcagc aacagcaaca
      661 accgccacca cctccccctc cacccccacc tcctcaactt cctcaacctc ctccacaggc
      721 acagcctctg ctgcctcagc cacaacctcc tccacctcca cctccacctc ctccaggccc
      781 agctgtggct gaggagcctc tgcaccgacc tggatccctg gtgagcaagg gcgaggagct
      841 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      901 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      961 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1021 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1081 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1141 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1201 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1261 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1321 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1381 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1441 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1501 cgggatcact ctcggcatgg acgagctgta caagtaatct agcgacctcg agtcatgtaa
     1561 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     1621 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     1681 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     1741 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     1801 ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     1861 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1921 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1981 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2041 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2101 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2161 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2221 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2281 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2341 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2401 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     2461 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agggtaataa
     2521 ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa
     2581 tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct
     2641 gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat
     2701 aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc
     2761 gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc
     2821 tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc
     2881 aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc
     2941 tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc
     3001 gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat
     3061 tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct
     3121 gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc
     3181 ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc
     3241 taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa
     3301 gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt
     3361 agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt
     3421 tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat
     3481 gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat
     3541 taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga
     3601 tgaattgaaa aggtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3661 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3721 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3781 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3841 aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc
     3901 gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt
     3961 tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa
     4021 aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag
     4081 acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca
     4141 caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct
     4201 gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta
     4261 catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa
     4321 tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag
     4381 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5221 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6301 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6361 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6421 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6481 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6541 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctctagta
//
