LOCUS       sequence-465750-        5125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Forms cytosolic HTT partial  exon 1 Q74 aggregates.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5125)
  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 5125)
  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: polyQ74-GFP
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1768..1777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1774..1776,1777..1779,1780..2493)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1774..1776,1777..1779,1780..2490)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1819..1840)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2080..2099)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2427..2448
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2616..2737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2653..2672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2707..2726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2744..3199)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2881..2902)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3093..3112
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3226..3330
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3332..3689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3332..3523
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3355..3375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3540..3675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3724..4518
                     /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             3724..4518
                     /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(3778..3797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4388..4407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4706..4725)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4750..4797
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctatg
     1321 gcgaccctgg aaaagctgat gaaggccttc gagtccctca agtccttcca gcagcagcag
     1381 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     1441 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     1501 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     1561 cagcagcaac agccgccacc gccgccgccg ccgccgccgc ctcctcagct tcctcagccg
     1621 ccgccgcagg cacagccgct gctgcctcag ccgcagccgc ccccgccgcc gcccccgccg
     1681 ccacccggcc cggctgtggc tgaggagccg ctgcaccgac caaagaaaga actttcagct
     1741 accaagaaag accgtgtagg atccatcgcc accatggtga gcaagggcga ggagctgttc
     1801 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1861 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1921 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1981 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2041 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2101 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2161 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2221 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2281 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2341 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2401 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2461 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2521 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2581 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2641 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2701 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2761 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2821 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2881 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2941 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3001 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3061 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3121 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3181 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3301 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3361 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3421 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3481 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3541 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3601 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3661 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3721 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3781 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3841 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3901 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3961 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4021 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4081 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4141 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4201 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4261 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4321 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4381 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4441 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4501 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4561 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4621 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4681 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4741 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4801 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4861 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4921 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5101 ccgtagaaaa gatcaaagga tcttc
//
