LOCUS       sequence-314037-        7476 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 7476)
  AUTHORS   Skinner PJ, Koshy BT, Cummings CJ, Klement IA, Helin K, Servadio A, 
            Zoghbi HY, Orr HT
  TITLE     Ataxin-1 with an expanded glutamine tract alters nuclear 
            matrix-associated structures.
  JOURNAL   Nature. 1997 Oct 30;389(6654):971-4.
  PUBMED    9353120
REFERENCE   2  (bases 1 to 7476)
  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: "Nature."; 
            date: "1997-10-30"; volume: "389(6654)"; pages: "971-4"
COMMENT     Sequence Label: pcDNA1 Flag ATXN1[2Q]
FEATURES             Location/Qualifiers
     source          1..7476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             792..815
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3150..3161
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(3435..3453)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3436..3453)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          3537..3602
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3732..3752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4024..4158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4061..4080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4115..4134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5009..5144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5639..5658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(5771..6229)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5908..5929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6123..6142
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(6353..6371)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6439..6543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6544..6612,6613..7404)
                     /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(6762..6781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcc gtgactttcc
      721 gtttatcaga ctaaaatcag agccatccag acagtgaaac agtcaccgtg gaggggggac
      781 ggcgaaaaat ggactacaaa gacgacgacg acaaaaaatc caaccaagag cggagcaacg
      841 aatgcctgcc tcccaagaag cgcgagatcc ccgccaccag ccggtcctcc gaggagaagg
      901 cccctaccct gcccagcgac aaccaccggg tggagggcac agcatggctc ccgggcaacc
      961 ctggtggccg gggccacggg ggcgggaggc atgggccggc agggacctcg gtggagcttg
     1021 gtttacaaca gggaataggt ttacacaaag cattgtccac agggctggac tactccccgc
     1081 ccagcgctcc caggtctgtc cccgtggcca ccacgctgcc tgccgcgtac gccaccccgc
     1141 agccagggac cccggtgtcc cccgtacagt acgctcacct gccgcacacc ttccagttca
     1201 ttgggtcctc ccaatacagt ggaacctatg ccagcttcat cccatcacag ctgatccccc
     1261 caaccgccaa ccccgtcacc agtgcagtgg cctcggccgc aggggccacc actccatccc
     1321 agcgctccca gctggaggcc tattccactc tgctggccaa catgggcagt ctgagccaga
     1381 cgccgggaca caaggctgag cagcagcacc tcagcagggc tccggggctt atcaccccgg
     1441 ggtccccccc accagcccag cagaaccagt acgtccacat ttccagttct ccgcagaaca
     1501 ccggccgcac cgcctctcct ccggccatcc ccgtccacct ccacccccac cagacgatga
     1561 tcccacacac gctcaccctg gggcccccct cccaggtcgt catgcaatac gccgactccg
     1621 gcagccactt tgtccctcgg gaggccacca agaaagctga gagcagccgg ctgcagcagg
     1681 ccatccaggc caaggaggtc ctgaacggtg agatggagaa gagccggcgg tacggggccc
     1741 cgtcctcagc cgacctgggc ctgggcaagg caggcggcaa gtcggttcct cacccgtacg
     1801 agtccaggca cgtggtggtc cacccgagcc cctcagacta cagcagtcgt gatccttcgg
     1861 gggtccgggc ctctgtgatg gtcctgccca acagcaacac gcccgcagct gacctggagg
     1921 tgcaacaggc cactcatcgt gaagcctccc cttctaccct caacgacaaa agtggcctgc
     1981 atttagggaa gcctggccac cggtcctacg cgctctcacc ccacacggtc attcagacca
     2041 cacacagtgc ttcagagcca ctcccggtgg gactgccagc cacggccttc tacgcaggga
     2101 ctcaaccccc tgtcatcggc tacctgagcg gccagcagca agcaatcacc tacgccggca
     2161 gcctgcccca gcacctggtg atccccggca cacagcccct gctcatcccg gtcggcagca
     2221 ctgacatgga agcgtcgggg gcagccccgg ccatagtcac gtcatccccc cagtttgctg
     2281 cagtgcctca cacgttcgtc accaccgccc ttcccaagag cgagaacttc aaccctgagg
     2341 ccctggtcac ccaggccgcc tacccagcca tggtgcaggc ccagatccac ctgcctgtgg
     2401 tgcagtccgt ggcctccccg gcggcggctc cccctacgct gcctccctac ttcatgaaag
     2461 gctccatcat ccagttggcc aacggggagc taaagaaggt ggaagactta aaaacagaag
     2521 atttcatcca gagtgcagag ataagcaacg acctgaagat cgactccagc accgtagaga
     2581 ggattgaaga cagccatagc ccgggcgtgg ccgtgataca gttcgccgtc ggggagcacc
     2641 gagcccaggt cagcgttgaa gttttggtag agtatccttt ttttgtgttt ggacagggct
     2701 ggtcatcctg ctgtccggag agaaccagcc agctctttga tttgccgtgt tccaaactct
     2761 cagttgggga tgtctgcatc tcgcttaccc tcaagaacct gaagaacggc tctgttaaaa
     2821 agggccagcc cgtggatccc gccagcgtcc tgctgaagca ctcaaaggcc gacggcctgg
     2881 cgggcagcag acacaggtat gccgagcagg aaaacggaat caaccagggg agtgcccaga
     2941 tgctctctga gaatggcgaa ctgaagtttc cagagaaaat gggattgcct gcagcgccct
     3001 tcctcaccaa aatagaaccc agcaagcccg cggcaacgag gaagaggagg tggtcggcgc
     3061 cagagagccg caaactggag aagtcagaag acgaaccacc tttgactctt cctaagcctt
     3121 ctctaattcc tcaggaggtt aagatttgca ttgaaggccg gtctaatgta ggcaagtaga
     3181 ggcagcgtgg gggaaaggaa acgtggctct cccttatcat ttgtatccag attactgtac
     3241 tgtaggctaa aataacacag tatttacatg ttatcttctt aattttaggt ttctgttcta
     3301 accttgtcat tagagttaca gcaggtgtgt cgcaggagac tggtgcatat gctttttcca
     3361 cgagtgtctg tcaggaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     3421 ctagagggcc ctattctata gtgtcaccta aatgctagag gatctttgtg aaggaacctt
     3481 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     3541 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     3601 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     3661 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     3721 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     3781 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     3841 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tatttgatgt
     3901 atagtgcctt gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct
     3961 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4021 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4081 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4141 tcttatcatg tctggatcga tcccgccatg gtatcaacgc catatttcta tttacagtag
     4201 ggacctcttc gttgtgtagg taccgctgta ttcctaggga aatagtagag gcaccttgaa
     4261 ctgtctgcat cagccatata gcccccgctg ttcgatttac aaacacaggc acagtactga
     4321 caaacccata cacctcctct gaaataccca tagttgctag ggctgtctcc gaactcatta
     4381 caccctccaa agtcagagct gtaatttcgc catcaagggc agcgagggct tctccagata
     4441 aaatagcttc tgccgagagt cccgtaaggg tagacacttc agctaatccc tcgatgaggt
     4501 ctactagaat agtcagtgcg gctcccattt tgaaaattca cttacttgat cagcttcaga
     4561 agatggcgga gggcctccaa cacagtaatt ttcctcccga ctcttaaaat agaaaatgtc
     4621 aagtcagtta agcaggaagt gactaactga ccgcagctgg ccgtgcgaca tcctctttta
     4681 attagttgct aggcaactgc cctccagagg gcagtgtggt tttgcaagag gaagcaaaaa
     4741 gcctctccac ccaggcctag aatgtttcca cccaatcatt actatgacaa cagctgtttt
     4801 ttttagtatt aagcagaggc cgggggcccc tggcctccgc ttactctgga gaaaaagaag
     4861 agaggcattg tagaggcttc cagaggcaac ttgtcaaaac aggactgctt ctatttctgt
     4921 cacactgtct ggccctgtca caaggtccag cacctccata ccccctttaa taagcagttt
     4981 gggaacgggt gcgggtctta ctccgcccat cccgccccta actccgccca gttccgccca
     5041 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5101 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5161 gctaattcgg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5341 tagcaccgcc tacatacctc gctctgctga agccagttac cagtggctgc tgccagtggc
     5401 gataagtcgt gtcttaccgg gttggactca agagatagtt accggataag gcgcagcggt
     5461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcaagctagc
     5761 ttctagctag aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5821 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5881 tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5941 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccg cccactacgt
     6001 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6061 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6121 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6181 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     6241 tttgacgagc acgtataacg tgctttcctc gttagaatca gagcgggagc taaacaggag
     6301 gccgattaaa gggattttag acaggaacgg tacgccagct ggacccgaaa gggcctcgtg
     6361 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6421 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6481 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6541 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6601 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6661 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6721 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6781 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6841 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6901 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6961 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7021 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7081 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7141 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7201 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7261 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7321 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7381 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7441 gatttaaaac ttcattttta atttctctag cgcgtt
//
