LOCUS       sequence-470436-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-EGFP with K408R mutation to block 
            SUMOylation.
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 (K408R)
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1449..1458
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1455..1457,1458..1460,1461..2174)
                     /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(1455..1457,1458..1460,1461..2171)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1500..1521)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1761..1780)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2108..2129
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2297..2418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2334..2353)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2388..2407
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2425..2880)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2562..2583)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2774..2793
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2907..3011
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3013..3370
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3013..3204
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3036..3056)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3221..3356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3405..4199
                     /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             3405..4199
                     /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(3459..3478)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4069..4088
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4387..4406)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4431..4478
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4807..5395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5296..5315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5570..5873
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5874..5945,1..132)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
       61 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      121 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
      181 agatgtctga atatattcgg gtaaccgaag atgagaacga tgagcccatt gaaataccat
      241 cggaagacga tgggacggtg ctgctctcca cggttacagc ccagtttcca ggggcgtgtg
      301 ggcttcgcta caggaatcca gtgtctcagt gtatgagagg tgtccggctg gtagaaggaa
      361 ttctgcatgc cccagatgct ggctggggaa atctggtgta tgttgtcaac tatccaaaag
      421 ataacaaaag aaaaatggat gagacagatg cttcatcagc agtgaaagtg aaaagagcag
      481 tccagaaaac atccgattta atagtgttgg gtctcccatg gaaaacaacc gaacaggacc
      541 tgaaagagta ttttagtacc tttggagaag ttcttatggt gcaggtcaag aaagatctta
      601 agactggtca ttcaaagggg tttggctttg ttcgttttac ggaatatgaa acacaagtga
      661 aagtaatgtc acagcgacat atgatagatg gacgatggtg tgactgcaaa cttcctaatt
      721 ctaagcaaag ccaagatgag cctttgagaa gcagaaaagt gtttgtgggg cgctgtacag
      781 aggacatgac tgaggatgag ctgcgggagt tcttctctca gtacggggat gtgatggatg
      841 tcttcatccc caagccattc agggcctttg cctttgttac atttgcagat gatcagattg
      901 cgcagtctct ttgtggagag gacttgatca ttaaaggaat cagcgttcat atatccaatg
      961 ccgaacctaa gcacaatagc aatagacagt tagaaagaag tggaagattt ggtggtaatc
     1021 caggtggctt tgggaatcag ggtggatttg gtaatagcag agggggtgga gctggtttgg
     1081 gaaacaatca aggtagtaat atgggtggtg ggatgaactt tggtgcgttc agcattaatc
     1141 cagccatgat ggctgccgcc caggcagcac tacagagcag ttggggtatg atgggcatgt
     1201 tagccagcca gcagaaccag tcaggcccat cgggtaataa ccaaaaccaa ggcaacatgc
     1261 agagggagcc aaaccaggcc ttcggttctg gaaataactc ttatagtggc tctaattctg
     1321 gtgcagcaat tggttgggga tcagcatcca atgcagggtc gggcagtggt tttaatggag
     1381 gctttggctc aagcatggat tctaggtctt ctggctgggg aatgggtacc gcgggcccgg
     1441 gatccatcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1501 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1561 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1621 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1681 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1741 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1801 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1861 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1921 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1981 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2041 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2101 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2161 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2221 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2281 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2341 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2401 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2461 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2521 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2581 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2641 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2701 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2761 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2821 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2881 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2941 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3001 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3061 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3121 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3181 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3241 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3301 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3361 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3421 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3481 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3541 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3601 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3661 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3721 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3781 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3841 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3901 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3961 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4021 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4081 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4141 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4201 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4261 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4321 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4381 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4441 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4501 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4561 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4621 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5461 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5521 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5581 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     5641 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     5701 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     5761 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     5821 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     5881 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     5941 tccac
//
