LOCUS       sequence-69310-e        7438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V591A and L686E mutated Ataxin1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7438)
  AUTHORS   Kim E, Lu HC, Zoghbi HY, Song JJ
  TITLE     Structural basis of protein complex formation and reconfiguration by
            polyglutamine disease protein Ataxin-1 and Capicua.
  JOURNAL   Genes Dev. 2013 Mar 15;27(6):590-5. doi: 10.1101/gad.212068.112.
  PUBMED    23512657
REFERENCE   2  (bases 1 to 7438)
  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: 
            "10.1101/gad.212068"; journalName: "Genes Dev"; date: "2013-03-15- 
            15"; volume: "27"; issue: "6"; pages: "590-5"
COMMENT     Sequence Label: Flag-ATXN1_V591A_L686E
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(603..1061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(740..761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     955..974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1186..1204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1272..1376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1377..1445,1446..2237)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1595..1614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2310..2689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2690..2893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2844..2864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2938..2957
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2938..2956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3101..3124
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5459..5470
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     intron          5812..5877
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6007..6027
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6299..6433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6336..6355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6390..6409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7279..7414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
       61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
      601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
      841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
     1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
     1141 aagggatttt agacaggaac ggtacgccag ctggattacc aaagggcctc gtgatacgcc
     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2281 aacttcattt ttaatttctc tagcgcgttg acattgatta ttgactagtt attaatagta
     2341 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2401 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2461 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
     2521 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2581 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2641 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2701 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2761 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2821 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2881 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     2941 tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc
     3001 gccagtgtgc tggaattccg tgactttccg tttatcagac taaaatcaga gccatccaga
     3061 cagtgaaaca gtcaccgtgg aggggggacg gcgaaaaatg gactacaaag acgacgacga
     3121 caaaaaatcc aaccaagagc ggagcaacga atgcctgcct cccaagaagc gcgagatccc
     3181 cgccaccagc cggtcctccg aggagaaggc ccctaccctg cccagcgaca accaccgggt
     3241 ggagggcaca gcatggctcc cgggcaaccc tggtggccgg ggccacgggg gcgggaggca
     3301 tgggccggca gggacctcgg tggagcttgg tttacaacag ggaataggtt tacacaaagc
     3361 attgtccaca gggctggact actccccgcc cagcgctccc aggtctgtcc ccgtggccac
     3421 cacgctgcct gccgcgtacg ccaccccgca gccagggacc ccggtgtccc ccgtacagta
     3481 cgctcacctg ccgcacacct tccagttcat tgggtcctcc caatacagtg gaacctatgc
     3541 cagcttcatc ccatcacagc tgatcccccc aaccgccaac cccgtcacca gtgcagtggc
     3601 ctcggccgca ggggccacca ctccatccca gcgctcccag ctggaggcct attccactct
     3661 gctggccaac atgggcagtc tgagccagac gccgggacac aaggctgagc agcagcacct
     3721 cagcagggct ccggggctca tcaccccggg gtccccccca ccagcccagc agaaccagta
     3781 cgtccacatt tccagttctc cgcagaacac cggccgcacc gcctctcctc cggccatccc
     3841 cgtccacctc cacccccacc agacgatgat cccacacacg ctcaccctgg ggcccccctc
     3901 ccaggtcgtc atgcaatacg ccgactccgg cagccacttt gtccctcggg aggccaccaa
     3961 gaaagctgag agcagccggc tgcagcaggc catccaggcc aaggaggtcc tgaacggtga
     4021 gatggagaag agccggcggt acggggcccc gtcctcagcc gacctgggcc tgggcaaggc
     4081 aggcggcaag tcggttcctc acccgtacga gtccaggcac gtggtggtcc acccgagccc
     4141 ctcagactac agcagtcgtg atccttcggg ggtccgggcc tctgtgatgg tcctgcccaa
     4201 cagcaacacg cccgcagctg acctggaggt gcaacaggcc actcatcgtg aagcctcccc
     4261 ttctaccctc aacgacaaaa gtggcctgca tttagggaag cctggccacc ggtcctacgc
     4321 gctctcaccc cacacggtca ttcagaccac acacagtgct tcagagccac tcccggtggg
     4381 actgccagcc acggccttct acgcagggac tcaaccccct gtcatcggct acctgagcgg
     4441 ccagcagcaa gcaatcacct acgccggcag cctgccccag cacctggtga tccccggcac
     4501 acagcccctg ctcatcccgg tcggcagcac tgacatggaa gcgtcggggg cagccccggc
     4561 catagtcacg tcatcccccc agtttgctgc agtgcctcac acgttcgtca ccaccgccct
     4621 tcccaagagc gagaacttca accctgaggc cctggtcacc caggccgcct acccagccat
     4681 ggtgcaggcc cagatccacc tgcctgtggt gcagtccgtg gcctccccgg cggcggctcc
     4741 ccctacgctg cctccctact tcatgaaagg ctccatcatc cagttggcca acggggagct
     4801 aaagaaggcg gaagacttaa aaacagaaga tttcatccag agtgcagaga taagcaacga
     4861 cctgaagatc gactccagca ccgtagagag gattgaagac agccatagcc cgggcgtggc
     4921 cgtgatacag ttcgccgtcg gggagcaccg agcccaggtc agcgttgaag ttttggtaga
     4981 gtatcctttt tttgtgtttg gacagggctg gtcatcctgc tgtccggaga gaaccagcca
     5041 gctctttgat ttgccgtgtt ccaaactctc agttggggat gtctgcatct cggaaaccct
     5101 caagaacctg aagaacggct ctgttaaaaa gggccagccc gtggatcccg ccagcgtcct
     5161 gctgaagcac tcaaaggccg acggcctggc gggcagcaga cacaggtatg ccgagcagga
     5221 aaacggaatc aaccagggga gtgcccagat gctctctgag aatggcgaac tgaagtttcc
     5281 agagaaaatg ggattgcctg cagcgccctt cctcaccaaa atagaaccca gcaagcccgc
     5341 ggcaacgagg aagaggaggt ggtcggcgcc agagagccgc aaactggaga agtcagaaga
     5401 cgaaccacct ttgactcttc ctaagccttc tctaattcct caggaggtta agatttgcat
     5461 tgaaggccgg tctaatgtag gcaagtagag gcagcgtggg ggaaaggaaa cgtggctctc
     5521 ccttatcatt tgtatccaga ttactgtact gtaggctaaa ataacacagt atttacatgt
     5581 tatcttctta attttaggtt tctgttctaa ccttgtcatt agagttacag caggtgtgtc
     5641 gcaggagact ggtgcatatg ctttttccac gagtgtctgt ctggaattct gcagatatcc
     5701 atcacactgg cggccgctcg agcatgcatc tagaggatct ttgtgaagga accttacttc
     5761 tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa ggtaaatata
     5821 aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt
     5881 ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg
     5941 ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct
     6001 actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta
     6061 agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc
     6121 acaaaggaaa aagctgcact gctatacaag aaaattatgg aaaaatattt gatgtatagt
     6181 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     6241 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     6301 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6361 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6421 tcatgtctgg atcatcccgc catggtatca acgccatatt tctatttaca gtagggacct
     6481 cttcgttgtg taggtaccgc tgtattccta gggaaatagt agaggcacct tgaactgtct
     6541 gcatcagcca tatagccccc gctgttcgac ttacaaacac aggcacagta ctgacaaacc
     6601 catacacctc ctctgaaata cccatagttg ctagggctgt ctccgaactc attacaccct
     6661 ccaaagtcag agctgtaatt tcgccatcaa gggcagcgag ggcttctcca gataaaatag
     6721 cttctgccga gagtcccgta agggtagaca cttcagctaa tccctcgatg aggtctacta
     6781 gaatagtcag tgcggctccc attttgaaaa ttcacttact tgatcagctt cagaagatgg
     6841 cggagggcct ccaacacagt aattttcctc ccgactctta aaatagaaaa tgtcaagtca
     6901 gttaagcagg aagtggacta actgacgcag ctggccgtgc gacatcctct tttaattagt
     6961 tgctaggcaa cgccctccag agggcgtgtg gttttgcaag aggaagcaaa agcctctcca
     7021 cccaggccta gaatgtttcc acccaatcat tactatgaca acagctgttt tttttagtat
     7081 taagcagagg ccggggaccc ctgggcccgc ttactctgga gaaaaagaag agaggcattg
     7141 tagaggcttc cagaggcaac ttgtcaaaac aggactgctt ctatttctgt cacactgtct
     7201 ggccctgtca caaggtccag cacctccata ccccctttaa taagcagttt gggaacgggt
     7261 gcgggtctta ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     7321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     7381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaattc
//
