LOCUS       sequence-377062-        5939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TDP-43 in mammalian cells. Note: EGFP and TDP-43 are 
            expressed in different reading frames..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5939)
  AUTHORS   Liu BW, Wang XY, Cao JL, Chen LL, Wang YL, Zhao BQ, Zhou J, Shen ZF
  TITLE     TDP-43 upregulates lipid metabolism modulator ABHD2 to suppress 
            apoptosis in hepatocellular carcinoma.
  JOURNAL   Commun Biol. 2022 Aug 13;5(1):816. doi: 10.1038/s42003-022-03788-w.
  PUBMED    35963893
REFERENCE   2  (bases 1 to 5939)
  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: "Commun 
            Biol."; date: "2022-08-13"; volume: "5(1):816. doi"; pages: " 
            10.1038/s42003-022-03788-w"
COMMENT     Sequence Label: pEGFP-TDP-43
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2727..2848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2818..2837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2855..3310)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3204..3223
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3337..3441
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3443..3800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3443..3634
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3466..3486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3651..3786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3835..4629
                     /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             3835..4629
                     /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(3889..3908)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4499..4518
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4817..4836)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4861..4908
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5237..5825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5726..5745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agatgtctga atatattcgg gtaaccgaag
     1381 atgagaacga tgagcccatt gaaataccat cggaagacga tgggacggtg ctgctctcca
     1441 cggttacagc ccagtttcca ggggcgtgtg ggcttcgcta caggaatcca gtgtctcagt
     1501 gtatgagagg tgtccggctg gtagaaggaa ttctgcatgc cccagatgct ggctggggaa
     1561 atctggtgta tgttgtcaac tatccaaaag ataacaaaag aaaaatggat gagacagatg
     1621 cttcatcagc agtgaaagtg aaaagagcag tccagaaaac atccgattta atagtgttgg
     1681 gtctcccatg gaaaacaacc gaacaggacc tgaaagagta ttttagtacc tttggagaag
     1741 ttcttatggt gcaggtcaag aaagatctta agactggtca ttcaaagggg tttggctttg
     1801 ttcgctttac ggaatatgaa acacaagtga aagtaatgtc acagcgacat atgatagatg
     1861 gacgatggtg tgactgcaaa cttcctaatt ctaagcaaag ccaagatgag cctttgagaa
     1921 gcagaaaagt gtttgtgggg cgctgtacag aggacatgac tgaggatgag ctgcgggagt
     1981 tcttctctca gtacggggat gtgatggatg tcttcatccc caagccattc agggcctttg
     2041 cctttgttac atttgcagac gatcagattg cgcagtctct ttgtggagag gacttgatca
     2101 ttaaaggaat cagcgttcat atatccaatg ccgaacctaa gcacaatagc aatagacagt
     2161 tagaaagaag tggaagattt ggtggtaatc caggtggctt tgggaatcag ggtggatttg
     2221 gtaatagcag agggggtgga gctggtttgg gaaacaatca aggtagtaat atgggtggtg
     2281 ggatgaactt tggtgcgttc agcattaatc cagccatgat ggctgccgcc caggcagcac
     2341 tacagagcag ttggggtatg atgggcatgt tagccagcca gcagaaccag tcaggcccat
     2401 cgggtaataa ccaaaaccaa ggcaacatgc agagggagcc aaaccaggcc ttcggttctg
     2461 gaaataactc ttatagtggc tctaattctg gtgcagcaat tggttgggga tcagcatcca
     2521 atgcagggtc gggcagtggt tttaatggag gctttggctc aagcatggat tctaagtctt
     2581 ctggctgggg aatgtaggga tccaccggat ctagataact gatcataatc agccatacca
     2641 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2701 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2821 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     2881 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3061 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3121 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3181 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3241 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3301 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3421 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3481 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3541 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3601 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3661 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3721 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3781 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4621 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4681 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4801 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4861 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4921 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4981 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5041 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5101 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5161 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5221 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5281 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5341 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5401 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5461 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5521 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5581 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5641 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5701 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5761 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5821 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5881 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
