LOCUS       sequence-181703-        5232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the GFP-tagged C-terminus of ataxin-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5232)
  AUTHORS   Pfeiffer A, Luijsterburg MS, Acs K, Wiegant WW, Helfricht A, Herzog 
            LK, Minoia M, Bottcher C, Salomons FA, van Attikum H, Dantuma NP
  TITLE     Ataxin-3 consolidates the MDC1-dependent DNA double-strand break 
            response by counteracting the SUMO-targeted ubiquitin ligase RNF4.
  JOURNAL   EMBO J. 2017 Mar 8. pii: e201695151. doi: 10.15252/embj.201695151.
  PUBMED    28275011
REFERENCE   2  (bases 1 to 5232)
  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: "EMBO J. 
            2017 Mar 8. pii: e201695151. doi: 10.15252/embj.201695151."
COMMENT     Sequence Label: GFP-ataxin-3 C-t
FEATURES             Location/Qualifiers
     source          1..5232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    128..249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(165..184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     219..238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(256..711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(393..414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     605..624
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        738..842
                     /gene="bla"
                     /label=AmpR promoter
     promoter        844..1201
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        844..1035
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(867..887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1052..1187
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1236..2030
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1236..2030
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1290..1309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1900..1919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2218..2237)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2262..2309
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2638..3226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3127..3146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3401..3704
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3705..3908
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3859..3879
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3947..3956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3953..3955,3956..3958,3959..4669)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3953..3955,3956..3958,3959..4669)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3998..4019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4259..4278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4606..4627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
       61 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
      121 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
      181 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
      241 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
      301 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      361 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      421 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      481 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
      541 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
      601 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
      661 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
      721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      841 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
      901 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1021 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1081 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1141 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1201 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     1261 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     1321 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     1381 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     1441 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     1501 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     1561 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     1621 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     1681 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1741 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     1801 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1861 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1921 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1981 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     2041 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     2101 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     2161 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     2221 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     2281 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2341 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2401 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2461 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     2521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3301 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     3361 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3421 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3481 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3541 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3601 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3661 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     3721 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3781 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     3841 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3901 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag
     3961 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4021 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4081 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4141 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4201 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4261 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4321 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4381 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4441 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4501 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4561 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4621 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     4681 atctcgagca gtccaacaga tgcatcgacc aaaacttatt ggagaagaat tagcacaact
     4741 aaaagagcaa agagtccata aaacagacct ggaacgagtg ttagaagcaa atgatggctc
     4801 aggaatgtta gacgaagatg aggaggattt gcagagggct ctggcactaa gtcgccaaga
     4861 aattgacatg gaagatgagg aagcagatct ccgcagggct attcagctaa gtatgcaagg
     4921 tagttccaga aacatatctc aagatatgac acagacatca ggtacaaatc ttacttcaga
     4981 agagcttcgg aagagacgag aagcctactt tgaaaaacag cagcaaaagc agcaacagca
     5041 gcagcagcag cagcagcagg gggacctatc aggacagagt tcacatccat gtgaaaggcc
     5101 agccaccagt tcaggagcac ttgggagtga tctaggtgat gctatgagtg aagaagacat
     5161 gcttcaggca gctgtgacca tgtctttaga aactgtcaga aatgatttga aaacagaagg
     5221 aaaaaaataa gg
//
