LOCUS       sequence-470433-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RNA-binding deficient TDP-43-EGFP .
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 (5FL)
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1052..1061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1058..1060,1061..1063,1064..1777)
                     /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(1058..1060,1061..1063,1064..1774)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1103..1124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1364..1383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1711..1732
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1900..2021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1937..1956)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1991..2010
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2028..2483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2186)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2377..2396
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2510..2614
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2616..2973
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2616..2807
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2639..2659)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2824..2959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3008..3802
                     /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             3008..3802
                     /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(3062..3081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3672..3691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3990..4009)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4034..4081
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4410..4998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4899..4918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5173..5476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5477..5680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5631..5651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtatgttgtc aactatccaa aagataacaa aagaaaaatg gatgagacag atgcttcatc
       61 agcagtgaaa gtgaaaagag cagtccagaa aacatccgat ttaatagtgt tgggtctccc
      121 atggaaaaca accgaacagg acctgaaaga gtatttaagt accttaggag aagttcttat
      181 ggtgcaggtc aagaaagatc ttaagactgg tcattcaaag gggttaggct tagttcgttt
      241 aacggaatat gaaacacaag tgaaagtaat gtcacagcga catatgatag atggacgatg
      301 gtgtgactgc aaacttccta attctaagca aagccaagat gagcctttga gaagcagaaa
      361 agtgtttgtg gggcgctgta cagaggacat gactgaggat gagctgcggg agttcttctc
      421 tcagtacggg gatgtgatgg atgtcttcat ccccaagcca ttcagggcct ttgcctttgt
      481 tacatttgca gatgatcaga ttgcgcagtc tctttgtgga gaggacttga tcattaaagg
      541 aatcagcgtt catatatcca atgccgaacc taagcacaat agcaatagac agttagaaag
      601 aagtggaaga tttggtggta atccaggtgg ctttgggaat cagggtggat ttggtaatag
      661 cagagggggt ggagctggtt tgggaaacaa tcaaggtagt aatatgggtg gtgggatgaa
      721 ctttggtgcg ttcagcatta atccagccat gatggctgcc gcccaggcag cactacagag
      781 cagttggggt atgatgggca tgttagccag ccagcagaac cagtcaggcc catcgggtaa
      841 taaccaaaac caaggcaaca tgcagaggga gccaaaccag gccttcggtt ctggaaataa
      901 ctcttatagt ggctctaatt ctggtgcagc aattggttgg ggatcagcat ccaatgcagg
      961 gtcgggcagt ggttttaatg gaggctttgg ctcaagcatg gattctaagt cttctggctg
     1021 gggaatgggt accgcgggcc cgggatccat cgccaccatg gtgagcaagg gcgaggagct
     1081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1261 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1501 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1561 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1621 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1681 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1741 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     1801 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1861 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1921 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1981 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2041 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2101 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2161 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2221 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2281 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2341 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2401 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2461 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2581 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2641 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2701 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2761 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2821 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2881 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2941 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3001 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3061 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3121 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3181 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3241 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3301 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3361 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3421 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3481 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3541 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3601 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3661 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3721 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3781 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3841 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3901 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3961 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4021 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4081 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4141 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4201 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4261 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5041 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5101 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5161 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5221 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5281 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5341 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5401 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5461 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5521 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5581 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5641 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     5701 ccgctagcgc taccggactc agatctcgag atgtctgaat atattcgggt aaccgaagat
     5761 gagaacgatg agcccattga aataccatcg gaagacgatg ggacggtgct gctctccacg
     5821 gttacagccc agtttccagg ggcgtgtggg cttcgctaca ggaatccagt gtctcagtgt
     5881 atgagaggtg tccggctggt agaaggaatt ctgcatgccc cagatgctgg ctggggaaat
     5941 ctggt
//
