LOCUS       sequence-471010-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-EGFP with phospho-dead mutations in the 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  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 5945)
  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 (4SA)
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..71
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(231..944,945..947,948..950))
                     /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             complement(join(234..944,945..947,948..950))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(276..297)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     625..644
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     884..905
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      947..956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2273..2476)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2302..2322)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2477..2780
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3035..3054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3872..3919
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3944..3963
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4151..4945)
                     /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             complement(4151..4945)
                     /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(4262..4281)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4872..4891
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4980..5337)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4994..5129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5146..5337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5294..5314
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5339..5443)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5470..5925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5557..5576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5767..5788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    join(5932..5945,1..108)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(5943..5945,1..17))
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
       61 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
      121 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
      181 aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct ttacttgtac
      241 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
      301 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
      361 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
      421 acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc
      481 ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg
      541 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
      601 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
      661 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
      721 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
      781 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
      841 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
      901 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgatg
      961 gatcccgggc ccgcggtacc cattccccag cctgctgcct tagaatccat agctgcgcca
     1021 aagcctccat taaaaccact gcccgaccct gcattggatg ctgatcccca accaattgct
     1081 gcaccagaat tagagccact ataagagtta tttccagaac cgaaggcctg gtttggctcc
     1141 ctctgcatgt tgccttggtt ttggttatta cccgatgggc ctgactggtt ctgctggctg
     1201 gctaacatgc ccatcatacc ccaactgctc tgtagtgctg cctgggcggc agccatcatg
     1261 gctggattaa tgctgaacgc accaaagttc atcccaccac ccatattact accttgattg
     1321 tttcccaaac cagctccacc ccctctgcta ttaccaaatc caccctgatt cccaaagcca
     1381 cctggattac caccaaatct tccacttctt tctaactgtc tattgctatt gtgcttaggt
     1441 tcggcattgg atatatgaac gctgattcct ttaatgatca agtcctctcc acaaagagac
     1501 tgcgcaatct gatcatctgc aaatgtaaca aaggcaaagg ccctgaatgg cttggggatg
     1561 aagacatcca tcacatcccc gtactgagag aagaactccc gcagctcatc ctcagtcatg
     1621 tcctctgtac agcgccccac aaacactttt ctgcttctca aaggctcatc ttggctttgc
     1681 ttagaattag gaagtttgca gtcacaccat cgtccatcta tcatatgtcg ctgtgacatt
     1741 actttcactt gtgtttcata ttccgtaaaa cgaacaaagc caaacccctt tgaatgacca
     1801 gtcttaagat ctttcttgac ctgcaccata agaacttctc caaaggtact aaaatactct
     1861 ttcaggtcct gttcggttgt tttccatggg agacccaaca ctattaaatc ggatgttttc
     1921 tggactgctc ttttcacttt cactgctgat gaagcatctg tctcatccat ttttcttttg
     1981 ttatcttttg gatagttgac aacatacacc agatttcccc agccagcatc tggggcatgc
     2041 agaattcctt ctaccagccg gacacctctc atacactgag acactggatt cctgtagcga
     2101 agcccacacg cccctggaaa ctgggctgta accgtggaga gcagcaccgt cccatcgtct
     2161 tccgatggta tttcaatggg ctcatcgttc tcatcttcgg ttacccgaat atattcagac
     2221 atctcgagat ctgagtccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc
     2281 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     2341 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     2401 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     2461 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     2521 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     2581 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     2641 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     2701 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     2761 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     2821 ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     3721 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     3781 gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     3841 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     3901 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     3961 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     4021 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     4081 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     4141 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4201 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4261 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4321 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     4381 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     4441 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4501 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4561 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4621 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4681 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4741 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     4801 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4861 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4921 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     4981 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     5041 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     5101 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     5161 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     5221 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     5281 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
     5341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5401 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5461 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5881 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcc ttaagataca
     5941 ttgat
//
