LOCUS       sequence-470421-        6783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-HA fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6783)
  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 6783)
  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: pDEST-TDP-43-HA
FEATURES             Location/Qualifiers
     source          1..6783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      116..704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     605..624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     858..875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    992..1013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1028..1033,1034..1051,1052..1058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1066..1082
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1071..1093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1090..1106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1090..1106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1155..1276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1166..1185)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1220..1239
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1452..2246)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1563..1582)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2173..2192
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2313..2642)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2327..2462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2381..2400)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2627..2647
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2656..3084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2978..2997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3130..3354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3343..3360
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3383..3400)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3410..3451)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    3477..3501
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3515..3541)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4797..4821)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4850..4869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4851..4869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4914..5117)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4943..4963)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5118..5497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5669..5688
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5763..5867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5868..5936,5937..6728)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6086..6105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
       61 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      901 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      961 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     1021 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1081 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     1141 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1201 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1261 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     1321 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     1381 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgcgaaccc
     1441 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     1501 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     1561 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     1621 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     1681 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     1741 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     1801 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     1861 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     1921 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     1981 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     2041 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     2101 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     2161 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     2221 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     2281 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
     2341 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     2401 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     2461 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     2521 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     2581 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     2641 acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2701 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2761 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2821 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2881 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2941 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3001 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3061 ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg
     3121 cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct
     3181 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     3241 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
     3301 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg
     3361 ctgatcagcg ggtttaaact caatggtgat ggtgatgatg accggtacgc gtagaatcga
     3421 gaccgaggag agggttaggg ataggcttac cttcgaaccg cgggccctct agatcaacca
     3481 ctttgtacaa gaaagctggg ttgcggccgc ttcaggcgta gtctggtacg tcgtatgggt
     3541 acattcccca gccagaagac ttagaatcca tgcttgagcc aaagcctcca ttaaaaccac
     3601 tgcccgaccc tgcattggat gctgatcccc aaccaattgc tgcaccagaa ttagagccac
     3661 tataagagtt atttccagaa ccgaaggcct ggtttggctc cctctgcatg ttgccttggt
     3721 tttggttatt acccgatggg cctgactggt tctgctggct ggctaacatg cccatcatac
     3781 cccaactgct ctgtagtgct gcctgggcgg cagccatcat ggctggatta atgctgaacg
     3841 caccaaagtt catcccacca cccatattac taccttgatt gtttcccaaa ccagctccac
     3901 cccctctgct attaccaaat ccaccctgat tcccaaagcc acctggatta ccaccaaatc
     3961 ttccacttct ttctaactgt ctattgctat tgtgcttagg ttcggcattg gatatatgaa
     4021 cgctgattcc tttaatgatc aagtcctctc cacaaagaga ctgcgcaatc tgatcatctg
     4081 caaatgtaac aaaggcaaag gccctgaatg gcttggggat gaagacatcc atcacatccc
     4141 cgtactgaga gaagaactcc cgcagctcat cctcagtcat gtcctctgta cagcgcccca
     4201 caaacacttt tctgcttctc aaaggctcat cttggctttg cttagaatta ggaagtttgc
     4261 agtcacacca tcgtccatct atcatatgtc gctgtgacat tactttcact tgtgtttcat
     4321 attccgtaaa acgaacaaag ccaaacccct ttgaatgacc agtcttaaga tctttcttga
     4381 cctgcaccat aagaacttct ccaaaggtac taaaatactc tttcaggtcc tgttcggttg
     4441 ttttccatgg gagacccaac actattaaat cggatgtttt ctggactgct cttttcactt
     4501 tcactgctga tgaagcatct gtctcatcca tttttctttt gttatctttt ggatagttga
     4561 caacatacac cagatttccc cagccagcat ctggggcatg cagaattcct tctaccagcc
     4621 ggacacctct catacactga gacactggat tcctgtagcg aagcccacac gcccctggaa
     4681 actgggctgt aaccgtggag agcagcaccg tcccatcgtc ttccgatggt atttcaatgg
     4741 gctcatcgtt ctcatcttcg gttacccgaa tatattcaga catctcgagc aagcttagcc
     4801 tgcttttttg tacaaacttg ttgatagctt aactagccag cttgggtctc cctatagtga
     4861 gtcgtattaa tttcgataag ccagtaagca gtgggttctc tagttagcca gagagctctg
     4921 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     4981 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     5041 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     5101 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     5161 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     5221 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     5281 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     5341 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     5401 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     5461 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
     5521 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     5581 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     5641 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5701 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     5761 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5821 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6781 ttt
//
