LOCUS       sequence-470435-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-EGFP with human ALS-causing Q331K 
            mutation.
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 (Q331K)
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..65
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1411..1420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1417..1419,1420..1422,1423..2136)
                     /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(1417..1419,1420..1422,1423..2133)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1462..1483)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1723..1742)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2070..2091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2259..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2296..2315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2350..2369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2387..2842)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2524..2545)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2736..2755
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2869..2973
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2975..3332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2975..3166
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2998..3018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3367..4161
                     /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             3367..4161
                     /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(3421..3440)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4031..4050
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4349..4368)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4393..4440
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4769..5357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5258..5277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5532..5835
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5836..5945,1..94)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
       61 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      121 gcgctaccgg actcagatct cgagatgtct gaatatattc gggtaaccga agatgagaac
      181 gatgagccca ttgaaatacc atcggaagac gatgggacgg tgctgctctc cacggttaca
      241 gcccagtttc caggggcgtg tgggcttcgc tacaggaatc cagtgtctca gtgtatgaga
      301 ggtgtccggc tggtagaagg aattctgcat gccccagatg ctggctgggg aaatctggtg
      361 tatgttgtca actatccaaa agataacaaa agaaaaatgg atgagacaga tgcttcatca
      421 gcagtgaaag tgaaaagagc agtccagaaa acatccgatt taatagtgtt gggtctccca
      481 tggaaaacaa ccgaacagga cctgaaagag tattttagta cctttggaga agttcttatg
      541 gtgcaggtca agaaagatct taagactggt cattcaaagg ggtttggctt tgttcgtttt
      601 acggaatatg aaacacaagt gaaagtaatg tcacagcgac atatgataga tggacgatgg
      661 tgtgactgca aacttcctaa ttctaagcaa agccaagatg agcctttgag aagcagaaaa
      721 gtgtttgtgg ggcgctgtac agaggacatg actgaggatg agctgcggga gttcttctct
      781 cagtacgggg atgtgatgga tgtcttcatc cccaagccat tcagggcctt tgcctttgtt
      841 acatttgcag atgatcagat tgcgcagtct ctttgtggag aggacttgat cattaaagga
      901 atcagcgttc atatatccaa tgccgaacct aagcacaata gcaatagaca gttagaaaga
      961 agtggaagat ttggtggtaa tccaggtggc tttgggaatc agggtggatt tggtaatagc
     1021 agagggggtg gagctggttt gggaaacaat caaggtagta atatgggtgg tgggatgaac
     1081 tttggtgcgt tcagcattaa tccagccatg atggctgccg cccaggcagc actaaagagc
     1141 agttggggta tgatgggcat gttagccagc cagcagaacc agtcaggccc atcgggtaat
     1201 aaccaaaacc aaggcaacat gcagagggag ccaaaccagg ccttcggttc tggaaataac
     1261 tcttatagtg gctctaattc tggtgcagca attggttggg gatcagcatc caatgcaggg
     1321 tcgggcagtg gttttaatgg aggctttggc tcaagcatgg attctaagtc ttctggctgg
     1381 ggaatgggta ccgcgggccc gggatccatc gccaccatgg tgagcaaggg cgaggagctg
     1441 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1501 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1561 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1621 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1681 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1741 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1801 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1861 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1921 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1981 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2041 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2101 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     2161 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2221 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2341 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2461 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2821 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2941 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3361 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3421 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3481 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3541 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3601 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3661 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3721 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3781 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3841 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3901 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3961 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4021 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4081 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4141 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4201 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4261 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4321 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4381 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4441 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4501 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4561 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5461 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     5521 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5581 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5641 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5701 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5761 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     5821 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     5881 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5941 caaaa
//
