LOCUS       sequence-470645-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-EGFP with K136R and K408R mutations.
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 (2KR)
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..53)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     244..263
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        377..481
                     /gene="bla"
                     /label=AmpR promoter
     promoter        483..840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        483..674
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(506..526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      691..826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             875..1669
                     /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             875..1669
                     /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(929..948)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1539..1558
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1857..1876)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1901..1948
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2277..2865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2766..2785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3040..3343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3344..3547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3498..3518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4864..4873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4870..4872,4873..4875,4876..5589)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVEMDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5176..5195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5523..5544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5712..5833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5749..5768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5803..5822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(5840..5945,1..350))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
       61 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      121 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      181 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      241 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      301 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
      361 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      421 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      481 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
      541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
      601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      661 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
      721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
      781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
      841 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
      901 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
      961 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     1021 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     1081 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     1141 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     1201 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     1261 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     1321 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1381 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     1441 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1501 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1561 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1621 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     1681 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     1741 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     1801 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     1861 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     1921 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     1981 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2041 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     2101 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     2161 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2221 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2281 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2341 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2401 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2461 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2521 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2581 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2641 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2701 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2761 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2821 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2881 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2941 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     3001 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     3061 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     3121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     3181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     3241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     3301 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     3361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     3421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     3481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     3541 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagatg
     3601 tctgaatata ttcgggtaac cgaagatgag aacgatgagc ccattgaaat accatcggaa
     3661 gacgatggga cggtgctgct ctccacggtt acagcccagt ttccaggggc gtgtgggctt
     3721 cgctacagga atccagtgtc tcagtgtatg agaggtgtcc ggctggtaga aggaattctg
     3781 catgccccag atgctggctg gggaaatctg gtgtatgttg tcaactatcc aaaagataac
     3841 aaaagaaaaa tggatgagac agatgcttca tcagcagtga aagtgaaaag agcagtccag
     3901 aaaacatccg atttaatagt gttgggtctc ccatggaaaa caaccgaaca ggacctgaaa
     3961 gagtatttta gtacctttgg agaagttctt atggtgcagg tcaggaaaga tcttaagact
     4021 ggtcattcaa aggggtttgg ctttgttcgt tttacggaat atgaaacaca agtgaaagta
     4081 atgtcacagc gacatatgat agatggacga tggtgtgact gcaaacttcc taattctaag
     4141 caaagccaag atgagccttt gagaagcaga aaagtgtttg tggggcgctg tacagaggac
     4201 atgactgagg atgagctgcg ggagttcttc tctcagtacg gggatgtgat ggatgtcttc
     4261 atccccaagc cattcagggc ctttgccttt gttacatttg cagatgatca gattgcgcag
     4321 tctctttgtg gagaggactt gatcattaaa ggaatcagcg ttcatatatc caatgccgaa
     4381 cctaagcaca atagcaatag acagttagaa agaagtggaa gatttggtgg taatccaggt
     4441 ggctttggga atcagggtgg atttggtaat agcagagggg gtggagctgg tttgggaaac
     4501 aatcaaggta gtaatatggg tggtgggatg aactttggtg cgttcagcat taatccagcc
     4561 atgatggctg ccgcccaggc agcactacag agcagttggg gtatgatggg catgttagcc
     4621 agccagcaga accagtcagg cccatcgggt aataaccaaa accaaggcaa catgcagagg
     4681 gagccaaacc aggccttcgg ttctggaaat aactcttata gtggctctaa ttctggtgca
     4741 gcaattggtt ggggatcagc atccaatgca gggtcgggca gtggttttaa tggaggcttt
     4801 ggctcaagca tggattctag gtcttctggc tggggaatgg gtaccgcggg cccgggatcc
     4861 atcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4921 gagatggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4981 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5041 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     5101 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5161 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5221 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5281 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5341 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5401 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5461 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5521 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5581 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     5641 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5701 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5761 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5821 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5881 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5941 tcaaa
//
