LOCUS       sequence-458380-        6478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast optimized human Htt (46Q) fused with aga2 protein gene under 
            gal1 promoter for the expression of Htt (46Q) in yeast surface 
            display. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6478)
  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 6478)
  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(46Q)-c_myc
FEATURES             Location/Qualifiers
     source          1..6478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      850..1438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1339..1358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1592..1609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1726..1747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1762..1767,1768..1785,1786..1792)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1800..1816
                     /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     1805..1827
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1824..1840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1824..1840
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1861..1879
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1909..2561
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2113..2230
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2525..2548
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2880..2891
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2892..2918
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3339..3368
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3697..3716)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3698..3716)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3723..3740)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3723..3739)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3732..3754)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3880..4335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4438..5112)
                     /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(5113..5393)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5414..5433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5533..5555
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5593..5611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5652..6155
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6192..6296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6297..6365,6366..6478,1..679)
                     /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 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
       61 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      121 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      181 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      241 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      301 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      361 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      421 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      481 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      541 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      601 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      661 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      721 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      961 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1501 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1561 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1621 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1681 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     1741 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     1801 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcga
     1861 aattaaccct cactaaaggg aacaaaagct ggtaccaatt ccttgaattt tcaaaaattc
     1921 ttactttttt tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac
     1981 catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta
     2041 tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc
     2101 tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct
     2161 cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg
     2221 cactgctccg aacaataaag attctacaat actagctttt atggttatga agaggaaaaa
     2281 ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga
     2341 tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat
     2401 ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt
     2461 tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat
     2521 tgttaatata cctctatact ttaacgtcaa ggagaaaaaa ccccggatcg aattccctac
     2581 ttcatacatt ttcaattaag atgcagttac ttcgctgttt ttcaatattt tctgttattg
     2641 cttcagtttt agcacaggaa ctgacaacta tatgcgagca aatcccctca ccaactttag
     2701 aatcgacgcc gtactctttg tcaacgacta ctattttggc caacgggaag gcaatgcaag
     2761 gagtttttga atattacaaa tcagtaacgt ttgtcagtaa ttgcggttct cacccctcaa
     2821 caactagcaa aggcagcccc ataaacacac agtatgtttt taaggacaat agctcgacga
     2881 ttgaaggtag atacccatac gacgttccag actacgctct gcaggctagt ggtggaggag
     2941 gctctggtgg aggcggtagc ggaggcggag ggtcgactag tatggcgacc ctggaaaagc
     3001 tgatgaaggc cttcgagtcc ctcaaaagct tccaacagca gcaacagcaa caacagcagc
     3061 aacagcaaca acagcagcaa cagcaacaac agcagcaaca gcagcaacag caacaacagc
     3121 agcaacagca acaacagcag caacagcaac aacagcagca acagcaacaa ccgccaccac
     3181 ctccccctcc acccccacct cctcaacttc ctcaacctcc tccacaggca cagcctctgc
     3241 tgcctcagcc acaacctcct ccacctccac ctccacctcc tccaggccca gctgtggctg
     3301 aggagcctct gcaccgacct ctcgaggggg gcggatccga acaaaagctt atttctgaag
     3361 aggacttgta atagagatct gataacaaca gtgtagatgt aacaaaatcg actttgttcc
     3421 cactgtactt ttagctcgta caaaatacaa tatacttttc atttctccgt aaacaacatg
     3481 ttttcccatg taatatcctt ttctattttt cgttccgtta ccaactttac acatacttta
     3541 tatagctatt cacttctata cactaaaaaa ctaagacaat tttaattttg ctgcctgcca
     3601 tatttcaatt tgttataaat tcctataatt tatcctatta gtagctaaaa aaagatgaat
     3661 gtgaatcgaa tcctaagaga attgagctcc aattcgccct atagtgagtc gtattacaat
     3721 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3781 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3841 cgcctttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4321 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4381 ccgcatagat cggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta
     4441 tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc
     4501 cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt
     4561 ttctggcgtc agtccaccag ctaacataaa atgtaagctt tcggggctct cttgccttcc
     4621 aacccagtca gaaatcgagt tccaatccaa aagttcacct gtcccacctg cttctgaatc
     4681 aaacaaggga ataaacgaat gaggtttctg tgaagctgca ctgagtagta tgttgcagtc
     4741 ttttggaaat acgagtcttt taataactgg caaaccgagg aactcttggt attcttgcca
     4801 cgactcatct ccatgcagtt ggacgatatc aatgccgtaa tcattgacca gagccaaaac
     4861 atcctcctta ggttgattac gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt
     4921 tttatatgct tttacaagac ttgaaatttt ccttgcaata accgggtcaa ttgttctctt
     4981 tctattgggc acacatataa tacccagcaa gtcagcatcg gaatctagag cacattctgc
     5041 ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga ccagaactac ctgtgaaatt
     5101 aataacagac atactccaag ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa
     5161 tagtcaccaa tgccctccct cttggccctc tccttttctt ttttcgaccg aattaattct
     5221 taatcggcaa aaaaagaaaa gctccggatc aagattgtac gtaaggtgac aagctatttt
     5281 tcaataaaga atatcttcca ctactgccat ctggcgtcat aactgcaaag tacacatata
     5341 ttacgatgct gttctattaa atgcttccta tattatatat atagtaatgt cgtgatctat
     5401 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5461 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5521 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5581 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5641 tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa
     5701 taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa
     5761 gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa
     5821 aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa
     5881 tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt
     5941 cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga
     6001 taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa
     6061 ctattttttc tttaatttct ttttttactt tctattttta atttatatat ttatattaaa
     6121 aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc
     6181 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6241 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccga
//
