LOCUS       sequence-461348-        4932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Forms nuclear HTT partial exon 1 Q74-NLS aggregates.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  AUTHORS   Korsten G, Osinga M, Pelle RA, Serweta AK, Hoogenberg B, Kampinga 
            HH, Kapitein LC
  TITLE     Nuclear poly-glutamine aggregates rupture the nuclear envelope and 
            hinder its repair.
  JOURNAL   J Cell Biol. 2024 Nov 4;223(11):e202307142. doi: 
            10.1083/jcb.202307142. Epub 2024 Aug 16.
  PUBMED    39150509
REFERENCE   2  (bases 1 to 4932)
  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: "J Cell 
            Biol."; date: "2024-11-4"; volume: "223(11):e202307142. doi"; pages:
            " 10.1083/jcb.202307142. Epub 2024 Aug 16"
COMMENT     Sequence Label: GFP-polyQ74-NLS
FEATURES             Location/Qualifiers
     source          1..4932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             108..128
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    273..394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(310..329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     364..383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(401..856)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(538..559)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     750..769
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        883..987
                     /gene="bla"
                     /label=AmpR promoter
     promoter        989..1346
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        989..1180
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1012..1032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1197..1332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1381..2175
                     /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             1381..2175
                     /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(1435..1454)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2045..2064
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2363..2382)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2407..2454
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2783..3371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3272..3291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3546..3849
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3850..4053
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4004..4024
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4092..4101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4098..4100,4101..4103,4104..4814)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4098..4100,4101..4103,4104..4814)
                     /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(4143..4164)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4404..4423)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4751..4772
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agcagcagca gcagcagcag cagcagcagc agcagcagca gcaacagccg ccaccgccgc
       61 cgccgccgcc gccgcctcct cagcttcctc agccgccgaa ttcgggccct aagaagaagc
      121 gaaaggttga ggatcctgct agcggatcca ccggatctag ataactgatc ataatcagcc
      181 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
      241 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
      301 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
      361 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
      421 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
      481 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
      541 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
      601 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
      661 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
      721 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
      781 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
      841 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      901 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      961 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     1021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     1201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     1261 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     1321 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     1381 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1441 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1501 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1561 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     1621 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     1681 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     1741 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     1801 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     1861 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     1921 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     1981 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2041 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2101 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2161 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     2221 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     2281 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     2341 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     2401 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     2461 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     2521 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     2581 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     2641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3421 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     3481 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     3541 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3601 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3661 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3721 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     3781 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     3841 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     3901 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     3961 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4021 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
     4081 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     4141 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     4201 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     4261 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     4321 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     4381 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     4441 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     4501 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     4561 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     4621 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     4681 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     4741 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     4801 acgagctgta caagtccgga ctcagatcta tgaaggcctt cgagtccctc aagtccttcc
     4861 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     4921 agcagcagca gc
//
