LOCUS       sequence-218128-        6816 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 6816)
  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 6816)
  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 103Q GAL
FEATURES             Location/Qualifiers
     source          1..6816
                     /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(1037..1039,1040..1753)
                     /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(1037..1039,1040..1750)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1079..1100)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1340..1359)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1687..1708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1770..2017
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1786..1804)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2035..2054)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2036..2054)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2064..2081)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2064..2080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2073..2095)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2221..2676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2308..2327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2518..2539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2807..3610)
                     /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(3611..3826)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3850..3869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3969..3991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4029..4047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4088..4591
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4628..4732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4733..4801,4802..5593)
                     /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(4951..4970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5764..6352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6253..6272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6506..6523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6640..6661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6676..6681,6682..6699,6700..6706)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6714..6730
                     /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     6719..6741
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6738..6754
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6738..6754
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6773..6793
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6775..6793
                     /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 acagcagcaa cagcaacaac agcagcaaca gcaacaacag cagcaacagc aacaacagca
      721 gcaacagcaa caacagcagc aacagcaaca acagcagcaa cagcaacaac agcagcaaca
      781 gcaacaacag cagcaacagc aacaacagca gcaacagcaa caacagcagc aacagcaaca
      841 acagcagcaa cagcaacaac agcagcaaca gcaacaaccg ccaccacctc cccctccacc
      901 cccacctcct caacttcctc aacctcctcc acaggcacag cctctgctgc ctcagccaca
      961 acctcctcca cctccacctc cacctcctcc aggcccagct gtggctgagg agcctctgca
     1021 ccgacctgga tccctggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1081 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1141 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1201 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1261 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1321 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1381 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1441 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1501 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1561 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1621 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1681 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1741 gctgtacaag taatctagtc gacctcgagt catgtaatta gttatgtcac gcttacattc
     1801 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1861 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     1921 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     1981 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     2041 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2101 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2161 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2221 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2281 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2341 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2401 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2461 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2521 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2581 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2641 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     2701 gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc
     2761 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     2821 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     2881 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     2941 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     3001 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     3061 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     3121 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     3181 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     3241 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     3301 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     3361 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     3421 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     3481 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     3541 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     3601 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     3661 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     3721 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     3781 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg
     3841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4021 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4081 ttagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
     4141 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
     4201 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
     4261 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
     4321 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
     4381 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
     4441 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
     4501 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
     4561 ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6541 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6601 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     6661 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     6721 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     6781 ccctcactaa agggaacaaa agctggagct ctagta
//
