LOCUS       sequence-181701-        5244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the GFP-tagged N-terminus (Josephin domain) of ataxin-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5244)
  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 5244)
  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 N-t
FEATURES             Location/Qualifiers
     source          1..5244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    124..245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(161..180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     215..234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(252..707)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(389..410)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     601..620
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        734..838
                     /gene="bla"
                     /label=AmpR promoter
     promoter        840..1197
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        840..1031
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(863..883)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1048..1183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1232..2026
                     /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             1232..2026
                     /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(1286..1305)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1896..1915
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2214..2233)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2258..2305
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2634..3222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3123..3142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3397..3700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3701..3904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3855..3875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3943..3952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3949..3951,3952..3954,3955..4665)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3949..3951,3952..3954,3955..4665)
                     /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(3994..4015)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4255..4274)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4602..4623
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
       61 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
      121 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
      181 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      241 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
      301 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      361 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
      421 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      481 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      541 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      601 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      661 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
      721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
      841 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
      901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
      961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1021 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1081 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     1141 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     1201 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     1261 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1321 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1381 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     1441 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1501 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1561 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1621 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1681 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1741 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     1801 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     1861 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     1921 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     1981 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     2041 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     2101 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     2161 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     2221 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2281 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     2341 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     2401 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     2461 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     2521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3301 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     3361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     3841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     3901 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag
     3961 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4021 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     4081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4141 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4201 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     4261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4381 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     4441 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     4501 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     4561 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4621 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
     4681 cgagctcaaa tggagtccat cttccacgag aaacaagaag gctcactttg tgctcaacat
     4741 tgcctgaata acttattgca aggagaatat tttagccctg tggaattatc ctcaattgca
     4801 catcagctgg atgaggagga gaggatgaga atggcagaag gaggagttac tagtgaagat
     4861 tatcgcacgt ttttacagca gccttctgga aatatggatg acagtggttt tttctctatt
     4921 caggttataa gcaatgcctt gaaagtttgg ggtttagaac taatcctgtt caacagtcca
     4981 gagtatcaga ggctcaggat cgatcctata aatgaaagat catttatatg caattataag
     5041 gaacactggt ttacagttag aaaattagga aaacagtggt ttaacttgaa ttctctcttg
     5101 acgggtccag aattaatatc agatacatat cttgcacttt tcttggctca attacaacag
     5161 gaaggttatt ctatatttgt cgttaagggt gatctgccag attgcgaagc tgaccaactc
     5221 ctgcagatga ttaggtgagg atcc
//
