LOCUS       sequence-470424-        5975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of chimeric human TDP-43-EGFP bearing a PY-nuclear 
            localization sequence (NLS) from HNRNPA1 in place of the native NLS 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5975)
  AUTHORS   Suk TR, Part CE, Zhang JL, Nguyen TT, Heer MM, Caballero-Gomez A, 
            Grybas VS, McKeever PM, Nguyen B, Ali T, Callaghan SM, Woulfe JM, 
            Robertson J, Rousseaux MWC
  TITLE     A stress-dependent TDP-43 SUMOylation program preserves neuronal 
            function.
  JOURNAL   Mol Neurodegener. 2025 Mar 28;20(1):38. doi: 
            10.1186/s13024-025-00826-z.
  PUBMED    40149017
REFERENCE   2  (bases 1 to 5975)
  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: "Mol 
            Neurodegener."; date: "2025-03-28"; volume: "20(1):38. doi"; pages: 
            " 10.1186/s13024-025-00826-z"
COMMENT     Sequence Label: pEGFP-TDP-43 (PY-NLS)
FEATURES             Location/Qualifiers
     source          1..5975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..245
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     196..216
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1592..1601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1598..1600,1601..1603,1604..2317)
                     /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(1598..1600,1601..1603,1604..2314)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1643..1664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1904..1923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2251..2272
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2440..2561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2477..2496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2531..2550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2568..3023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2705..2726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2917..2936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3050..3154
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3156..3513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3156..3347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3179..3199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3364..3499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3548..4342
                     /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             3548..4342
                     /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(3602..3621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4212..4231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4530..4549)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4574..4621
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4950..5538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5439..5458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5713..5975,1..41)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
       61 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      121 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      181 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      241 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagatgtc
      301 tgaatatatt cgggtaaccg aagatgagaa cgatgagccc attgaaatac catcggaaga
      361 cgatgggacg gtgctgctct ccacggttac agcccagttt ccaggggcgt gtgggcttcg
      421 ctacaggaat ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag gaattctgca
      481 tgccccagat gctggctggg gaaatctggt gtatgttgtc aactatccaa aagataactt
      541 tggtaactac aacaaccagt cctcaaactt tgggccgatg agaggcggaa atttcggcgg
      601 acgctccagc ggaccgtatg cagtccagaa aacatccgat ttaatagtgt tgggtctccc
      661 atggaaaaca accgaacagg acctgaaaga gtattttagt acctttggag aagttcttat
      721 ggtgcaggtc aagaaagatc ttaagactgg tcattcaaag gggtttggct ttgttcgttt
      781 tacggaatat gaaacacaag tgaaagtaat gtcacagcga catatgatag atggacgatg
      841 gtgtgactgc aaacttccta attctaagca aagccaagat gagcctttga gaagcagaaa
      901 agtgtttgtg gggcgctgta cagaggacat gactgaggat gagctgcggg agttcttctc
      961 tcagtacggg gatgtgatgg atgtcttcat ccccaagcca ttcagggcct ttgcctttgt
     1021 tacatttgca gatgatcaga ttgcgcagtc tctttgtgga gaggacttga tcattaaagg
     1081 aatcagcgtt catatatcca atgccgaacc taagcacaat agcaatagac agttagaaag
     1141 aagtggaaga tttggtggta atccaggtgg ctttgggaat cagggtggat ttggtaatag
     1201 cagagggggt ggagctggtt tgggaaacaa tcaaggtagt aatatgggtg gtgggatgaa
     1261 ctttggtgcg ttcagcatta atccagccat gatggctgcc gcccaggcag cactacagag
     1321 cagttggggt atgatgggca tgttagccag ccagcagaac cagtcaggcc catcgggtaa
     1381 taaccaaaac caaggcaaca tgcagaggga gccaaaccag gccttcggtt ctggaaataa
     1441 ctcttatagt ggctctaatt ctggtgcagc aattggttgg ggatcagcat ccaatgcagg
     1501 gtcgggcagt ggttttaatg gaggctttgg ctcaagcatg gattctaagt cttctggctg
     1561 gggaatgggt accgcgggcc cgggatccat cgccaccatg gtgagcaagg gcgaggagct
     1621 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1681 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1741 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1801 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1861 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1921 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1981 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2041 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2101 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2161 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     2221 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2281 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     2341 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2401 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2521 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2581 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2641 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2701 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2761 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2821 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2881 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2941 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3001 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3061 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3121 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3361 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3421 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3481 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3541 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3601 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3661 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3721 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3781 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3841 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3901 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3961 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4021 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4081 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4141 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4201 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4261 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4321 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4381 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4441 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4501 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4561 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4621 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4681 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4741 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4801 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4861 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5581 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5641 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5701 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5761 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5821 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5881 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5941 ttatgcccag tacatgacct tatgggactt tccta
//
