LOCUS       sequence-448162-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP tagged upsteram ORF of NOTCH2NLC with a control size 
            (12x) of GGC repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  AUTHORS   Boivin M, Deng J, Pfister V, Grandgirard E, Oulad-Abdelghani M, 
            Morlet B, Ruffenach F, Negroni L, Koebel P, Jacob H, Riet F, 
            Dijkstra AA, McFadden K, Clayton WA, Hong D, Miyahara H, Iwasaki Y, 
            Sone J, Wang Z, Charlet-Berguerand N
  TITLE     Translation of GGC repeat expansions into a toxic polyglycine 
            protein in NIID defines a novel class of human genetic disorders: 
            The polyG diseases.
  JOURNAL   Neuron. 2021 Jun 2;109(11):1825-1835.e5. doi: 
            10.1016/j.neuron.2021.03.038. Epub 2021 Apr 21.
  PUBMED    33887199
REFERENCE   2  (bases 1 to 5555)
  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"; doi: 
            "10.1016/j.neuron.2021.03.038"; journalName: "Neuron"; date: 
            "2021-06-2- 2"; volume: "109"; issue: "11"; pages: "1825-1835.e5"
COMMENT     Sequence Label: pAAV uN2C GFP (Ctl)
FEATURES             Location/Qualifiers
     source          1..5555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             join(1502..1504,1505..2218)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1502..1504,1505..2215)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1805..1824)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2152..2173
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2261..2287
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2302..2778
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2501..2520)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2818..2958
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2818..2947
                     /label=AAV2 ITR
     rep_origin      3033..3488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3120..3139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3330..3351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3505..3524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3624..3646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3684..3702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3770..3874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3875..3943,3944..4735)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4093..4112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4906..5494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5395..5414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc gaacatcgat tgaattcaaa
     1321 gcttgccatg tggatctgcc caggaggagg aggaggagga ggcggcggcg gcggaggcgg
     1381 cgaccgagaa gatgcccgcc ctgcgccgct ctgctgtggg cgctgctggc gctctggctg
     1441 tgctgcgcga cccccgcgca tgcattgcag tgtcgagatg gctatgaacc ctgtgggatc
     1501 cgtcagcaaa ggcgaggagc ttttcaccgg ggttgttccc atcctcgtcg agctcgacgg
     1561 cgacgtaaac ggccacaaat tcagcgtttc cggcgagggc gagggcgatg ccacctacgg
     1621 caagctcacc ctcaagttca tctgcaccac cggcaagctc cccgtcccct ggcccaccct
     1681 cgtcaccacc ctcacctacg gcgtccagtg cttcagccgc taccccgacc acatgaagca
     1741 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1801 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1861 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1921 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1981 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2041 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2101 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2161 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgata
     2221 agaattctgc agatatccag cacagtggcg gccgctcgag tacccatacg atgttccaga
     2281 ttacgctaag taaagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     2341 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     2401 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     2461 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     2521 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     2581 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     2641 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     2701 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     2761 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     2821 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2881 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2941 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3001 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3061 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3121 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3181 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3241 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3301 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3361 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3421 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3481 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3601 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3661 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3721 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3841 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3901 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3961 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4021 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4081 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4141 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4201 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4261 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4321 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4381 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4441 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4501 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4561 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4621 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4681 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4741 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5041 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5521 gttcctggcc ttttgctggc cttttgctca catgt
//
