LOCUS       sequence-314860-        7725 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 7725)
  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 7725)
  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[85Q]
FEATURES             Location/Qualifiers
     source          1..7725
                     /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             3399..3410
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(3684..3702)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3685..3702)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          3786..3851
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3981..4001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4273..4407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4364..4383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5258..5393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5888..5907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(6020..6478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6157..6178)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6372..6391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(6602..6620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6688..6792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6793..6861,6862..7653)
                     /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(7011..7030)
                     /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 cagcagcagc agcagcagca gcagcagcag cagcagcagc
     1441 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     1501 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     1561 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     1621 agcagcagca gcagcagcag cagcagcagc agcagcacct cagcagggct ccggggctca
     1681 tcaccccggg gtccccccca ccagcccagc agaaccagta cgtccacatt tccagttctc
     1741 cgcagaacac cggccgcacc gcctctcctc cggccatccc cgtccacctc cacccccacc
     1801 agacgatgat cccacacacg ctcaccctgg ggcccccctc ccaggtcgtc atgcaatacg
     1861 ccgactccgg cagccacttt gtccctcggg aggccaccaa gaaagccgag agcagccggc
     1921 tgcagcaggc catccaggcc aaggaggtcc tgaacggtga gatggagaag agccggcggt
     1981 acggggcccc gtcctcagcc gacctgggcc tgggcaaggc aggcggcaag tcggttcctc
     2041 acccgtacga gtccaggcac gtggtggtcc acccgagccc ctcagactac agcagtcgtg
     2101 atccttcggg ggtccgggcc tctgtgatgg tcctgcccaa cagcaacacg cccgcagctg
     2161 acctggaggt gcaacaggcc actcatcgtg aagcctcccc ttctaccctc aacgacaaaa
     2221 gtggcctgca tttagggaag cctggccacc ggtcctacgc gctctcaccc cacacggtca
     2281 ttcagaccac acacagtgct tcagagccac tcccggtggg actgccagcc acggccttct
     2341 acgcagggac tcaaccccct gtcatcggct acctgagcgg ccagcagcaa gcaatcacct
     2401 acgccggcag cctgccccag cacctggtga tccccggcac acagcccctg ctcatcccgg
     2461 tcggcagcac tgacatggaa gcgtcggggg cagccccggc catagtcacg tcatcccccc
     2521 agtttgctgc agtgcctcac acgttcgtca ccaccgccct tcccaagagc gagaacttca
     2581 accctgaggc cctggtcacc caggccgcct acccagccat ggtgcaggcc cagatccacc
     2641 tgcctgtggt gcagtccgtg gcctccccgg cggcggctcc ccctacgctg cctccctact
     2701 tcatgaaagg ctccatcatc cagttggcca acggggagct aaagaaggtg gaagacttaa
     2761 aaacagaaga tttcatccag agtgcagaga taagcaacga cctgaagatc gactccagca
     2821 ccgtagagag gattgaagac agccatagcc cgggcgtggc cgtgatacag ttcgccgtcg
     2881 gggagcaccg agcccaggtc agcgttgaag ttttggtaga gtatcctttt tttgtgtttg
     2941 gacagggctg gtcatcctgc tgtccggaga gaaccagcca gctctttgat ttgccgtgtt
     3001 ccaaactctc agttggggat gtctgcatct cgcttaccct caagaacctg aagaacggct
     3061 ctgttaaaaa gggccagccc gtggatcccg ccagcgtcct gctgaagcac tcaaaggccg
     3121 acggcctggc gggcagcaga cacaggtatg ccgagcagga aaacggaatc aaccagggga
     3181 gtgcccagat gctctctgag aatggcgaac tgaagtttcc agagaaaatg ggattgcctg
     3241 cagcgccctt cctcaccaaa atagaaccca gcaagcccgc ggcaacgagg aagaggaggt
     3301 ggtcggcgcc agagagccgc aaactggaga agtcagaaga cgaaccacct ttgactcttc
     3361 ctaagccttc tctaattcct caggaggtta agatttgcat tgaaggccgg tctaatgtag
     3421 gcaagtagag gcagcgtggg ggaaaggaaa cgtggctctc ccttatcatt tgtatccaga
     3481 ttactgtact gtaggctaaa ataacacagt atttacatgt tatcttctta attttaggtt
     3541 tctgttctaa ccttgtcatt agagttacag caggtgtgtc gcaggagact ggtgcatatg
     3601 ctttttccac gagtgtctgt caggaattct gcagatatcc atcacactgg cggccgctcg
     3661 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagg atctttgtga
     3721 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
     3781 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     3841 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     3901 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     3961 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     4021 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     4081 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     4141 atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt
     4201 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     4261 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4321 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4381 atcaatgtat cttatcatgt ctggatcgat cccgccatgg tatcaacgcc atatttctat
     4441 ttacagtagg gacctcttcg ttgtgtaggt accgctgtat tcctagggaa atagtagagg
     4501 caccttgaac tgtctgcatc agccatatag cccccgctgt tcgatttaca aacacaggca
     4561 cagtactgac aaacccatac acctcctctg aaatacccat agttgctagg gctgtctccg
     4621 aactcattac accctccaaa gtcagagctg taatttcgcc atcaagggca gcgagggctt
     4681 ctccagataa aatagcttct gccgagagtc ccgtaagggt agacacttca gctaatccct
     4741 cgatgaggtc tactagaata gtcagtgcgg ctcccatttt gaaaattcac ttacttgatc
     4801 agcttcagaa gatggcggag ggcctccaac acagtaattt tcctcccgac tcttaaaata
     4861 gaaaatgtca agtcagttaa gcaggaagtg actaactgac cgcagctggc cgtgcgacat
     4921 cctcttttaa ttagttgcta ggcaactgcc ctccagaggg cagtgtggtt ttgcaagagg
     4981 aagcaaaaag cctctccacc caggcctaga atgtttccac ccaatcatta ctatgacaac
     5041 agctgttttt tttagtatta agcagaggcc gggggcccct ggcctccgct tactctggag
     5101 aaaaagaaga gaggcattgt agaggcttcc agaggcaact tgtcaaaaca ggactgcttc
     5161 tatttctgtc acactgtctg gccctgtcac aaggtccagc acctccatac cccctttaat
     5221 aagcagtttg ggaacgggtg cgggtcttac tccgcccatc ccgcccctaa ctccgcccag
     5281 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5341 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5401 ttgcaaaaag ctaattcggc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5521 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5581 agaactctgt agcaccgcct acatacctcg ctctgctgaa gccagttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gagatagtta ccggataagg
     5701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6001 caagctagct tctagctaga aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     6061 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6121 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6181 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccgc
     6241 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     6301 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6361 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6421 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     6481 tatggttgct ttgacgagca cgtataacgt gctttcctcg ttagaatcag agcgggagct
     6541 aaacaggagg ccgattaaag ggattttaga caggaacggt acgccagctg gacccgaaag
     6601 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6661 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6781 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6841 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6901 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6961 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7021 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7081 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7141 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7201 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7261 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7321 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7381 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7441 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7501 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7561 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7621 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7681 ctttagattg atttaaaact tcatttttaa tttctctagc gcgtt
//
