LOCUS       sequence-458382-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast optimized human Htt (25Q) fused with aga2 protein gene under 
            gal1 promoter for the expression of Htt (25Q) in yeast surface 
            display. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  AUTHORS   Khan A, Ozcelik CE, Begli O, Oguz O, Kesici MS, Kasirga TS, 
            Ozcubukcu S, Yuca E, Seker UOS
  TITLE     Highly Potent Peptide Therapeutics To Prevent Protein Aggregation in
            Huntington's Disease.
  JOURNAL   ACS Med Chem Lett. 2023 Nov 14;14(12):1821-1826. doi: 
            10.1021/acsmedchemlett.3c00415. eCollection 2023 Dec 14.
  PUBMED    38116434
REFERENCE   2  (bases 1 to 6415)
  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"; doi: 
            "10.1021/acsmedchemlett.3c00415"; journalName: "ACS Med Chem Lett"; 
            date: "2023-11-14- 14"; volume: "14"; issue: "12"; pages: 
            "1821-1826"
COMMENT     Sequence Label: pETcon-pGAL1-Aga2-Htt(25Q)-c_myc
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      842..1430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1331..1350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1584..1601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1718..1739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1754..1759,1760..1777,1778..1784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1792..1808
                     /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     1797..1819
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1816..1832
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1816..1832
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1853..1871
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1901..2553
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2105..2222
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2517..2540
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2872..2883
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2884..2910
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3268..3297
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3626..3645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3627..3645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3652..3669)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3652..3668)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3661..3683)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3809..4264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4106..4127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4367..5041)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5042..5322)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5343..5362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5462..5484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5522..5540)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5581..6084
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6121..6225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6226..6294,6295..6415,1..671)
                     /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"
ORIGIN
        1 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
       61 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      121 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      181 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      241 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
      301 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
      361 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
      421 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
      481 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
      541 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
      601 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
      661 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
      721 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
      781 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
      841 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
      901 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      961 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1021 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1081 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1141 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1201 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1261 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1321 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1381 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1441 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1501 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     1561 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     1621 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1681 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     1741 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     1801 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc
     1861 ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat tcttactttt
     1921 tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc accatataca
     1981 tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat tatcttagcc
     2041 taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt gctatattga
     2101 agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact ctcctccgtg
     2161 cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc
     2221 cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt
     2281 aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag gatgataatg
     2341 cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc
     2401 tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac tttcaacatt
     2461 ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa attgttaata
     2521 tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct acttcataca
     2581 ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat tgcttcagtt
     2641 ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt agaatcgacg
     2701 ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca aggagttttt
     2761 gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc aacaactagc
     2821 aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac gattgaaggt
     2881 agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg aggctctggt
     2941 ggaggcggta gcggaggcgg agggtcgact agtatggcga ccctggaaaa gctgatgaag
     3001 gccttcgagt ccctcaaaag cttccaacag cagcaacagc aacaacagca gcaacagcaa
     3061 caacagcagc aacagcaaca acagcagcaa cagcaacaac cgccaccacc tccccctcca
     3121 cccccacctc ctcaacttcc tcaacctcct ccacaggcac agcctctgct gcctcagcca
     3181 caacctcctc cacctccacc tccacctcct ccaggcccag ctgtggctga ggagcctctg
     3241 caccgacctc tcgagggggg cggatccgaa caaaagctta tttctgaaga ggacttgtaa
     3301 tagagatctg ataacaacag tgtagatgta acaaaatcga ctttgttccc actgtacttt
     3361 tagctcgtac aaaatacaat atacttttca tttctccgta aacaacatgt tttcccatgt
     3421 aatatccttt tctatttttc gttccgttac caactttaca catactttat atagctattc
     3481 acttctatac actaaaaaac taagacaatt ttaattttgc tgcctgccat atttcaattt
     3541 gttataaatt cctataattt atcctattag tagctaaaaa aagatgaatg tgaatcgaat
     3601 cctaagagaa ttgagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt
     3661 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3721 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcctttccca
     3781 acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg
     3841 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3901 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3961 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4021 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4081 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4141 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4201 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4261 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagatc
     4321 ggcaagtgca caaacaatac ttaaataaat actactcagt aataacctat ttcttagcat
     4381 ttttgacgaa atttgctatt ttgttagagt cttttacacc atttgtctcc acacctccgc
     4441 ttacatcaac accaataacg ccatttaatc taagcgcatc accaacattt tctggcgtca
     4501 gtccaccagc taacataaaa tgtaagcttt cggggctctc ttgccttcca acccagtcag
     4561 aaatcgagtt ccaatccaaa agttcacctg tcccacctgc ttctgaatca aacaagggaa
     4621 taaacgaatg aggtttctgt gaagctgcac tgagtagtat gttgcagtct tttggaaata
     4681 cgagtctttt aataactggc aaaccgagga actcttggta ttcttgccac gactcatctc
     4741 catgcagttg gacgatatca atgccgtaat cattgaccag agccaaaaca tcctccttag
     4801 gttgattacg aaacacgcca accaagtatt tcggagtgcc tgaactattt ttatatgctt
     4861 ttacaagact tgaaattttc cttgcaataa ccgggtcaat tgttctcttt ctattgggca
     4921 cacatataat acccagcaag tcagcatcgg aatctagagc acattctgcg gcctctgtgc
     4981 tctgcaagcc gcaaactttc accaatggac cagaactacc tgtgaaatta ataacagaca
     5041 tactccaagc tgcctttgtg tgcttaatca cgtatactca cgtgctcaat agtcaccaat
     5101 gccctccctc ttggccctct ccttttcttt tttcgaccga attaattctt aatcggcaaa
     5161 aaaagaaaag ctccggatca agattgtacg taaggtgaca agctattttt caataaagaa
     5221 tatcttccac tactgccatc tggcgtcata actgcaaagt acacatatat tacgatgctg
     5281 ttctattaaa tgcttcctat attatatata tagtaatgtc gtgatctatg gtgcactctc
     5341 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5401 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5461 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5521 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5581 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
     5641 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
     5701 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
     5761 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
     5821 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
     5881 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
     5941 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
     6001 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
     6061 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
     6121 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6181 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6241 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6301 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6361 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgtt
//
