LOCUS       sequence-470419-        7030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TDP-43-SUMO2-HA fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  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 7030)
  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-SUMO2-HA
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     377..396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(615..719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(794..813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        985..1364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1365..1568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1519..1539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1613..1632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1613..1631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1661..1685
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3198..3224
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3228..3252)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3278..3319
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3329..3346
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3369..3386)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3375..3599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3645..4073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3732..3751)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3942..3963
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4082..4102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4087..4416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4267..4402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4329..4348
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4483..5277
                     /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(4537..4556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5147..5166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5453..5574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5490..5509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5544..5563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5623..5639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5623..5639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5636..5658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5647..5663
                     /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)."
     promoter        complement(join(5671..5677,5678..5695,5696..5701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5716..5737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5854..5871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6025..6613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6105..6124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6784..7030,1..545,546..614))
                     /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"
ORIGIN
        1 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
       61 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      721 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
      781 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
      841 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
      901 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      961 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     1021 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1081 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1141 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1201 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1261 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1321 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1381 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1441 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1501 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1561 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     1621 ctcactatag ggagacccaa gctggctagt taagctatca acaagtttgt acaaaaaagc
     1681 aggctatgtc tgaatatatt cgggtaaccg aagatgagaa cgatgagccc attgaaatac
     1741 catcggaaga cgatgggacg gtgctgctct ccacggttac agcccagttt ccaggggcgt
     1801 gtgggcttcg ctacaggaat ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag
     1861 gaattctgca tgccccagat gctggctggg gaaatctggt gtatgttgtc aactatccaa
     1921 aagataacaa aagaaaaatg gatgagacag atgcttcatc agcagtgaaa gtgaaaagag
     1981 cagtccagaa aacatccgat ttaatagtgt tgggtctccc atggaaaaca accgaacagg
     2041 acctgaaaga gtattttagt acctttggag aagttcttat ggtgcaggtc aagaaagatc
     2101 ttaagactgg tcattcaaag gggtttggct ttgttcgttt tacggaatat gaaacacaag
     2161 tgaaagtaat gtcacagcga catatgatag atggacgatg gtgtgactgc aaacttccta
     2221 attctaagca aagccaagat gagcctttga gaagcagaaa agtgtttgtg gggcgctgta
     2281 cagaggacat gactgaggat gagctgcggg agttcttctc tcagtacggg gatgtgatgg
     2341 atgtcttcat ccccaagcca ttcagggcct ttgcctttgt tacatttgca gatgatcaga
     2401 ttgcgcagtc tctttgtgga gaggacttga tcattaaagg aatcagcgtt catatatcca
     2461 atgccgaacc taagcacaat agcaatagac agttagaaag aagtggaaga tttggtggta
     2521 atccaggtgg ctttgggaat cagggtggat ttggtaatag cagagggggt ggagctggtt
     2581 tgggaaacaa tcaaggtagt aatatgggtg gtgggatgaa ctttggtgcg ttcagcatta
     2641 atccagccat gatggctgcc gcccaggcag cactacagag cagttggggt atgatgggca
     2701 tgttagccag ccagcagaac cagtcaggcc catcgggtaa taaccaaaac caaggcaaca
     2761 tgcagaggga gccaaaccag gccttcggtt ctggaaataa ctcttatagt ggctctaatt
     2821 ctggtgcagc aattggttgg ggatcagcat ccaatgcagg gtcgggcagt ggttttaatg
     2881 gaggctttgg ctcaagcatg gattctaagt cttctggctg gggaatgatg gacgaaaagc
     2941 ccaaggaagg agtcaagact gagaacaacg atcatattaa tttgaaggtg gcggggcagg
     3001 atggttctgt ggtgcagttt aagattaaga ggcatacacc acttagtaaa ctaatgaaag
     3061 cctattgtga acgacaggga ttgtcaatga ggcagatcag attccgattt gacgggcaac
     3121 caatcaatga aacagacaca cctgcacagt tggaaatgga ggatgaagat acaattgatg
     3181 tgttccaaca gcagacgtac ccatacgacg taccagacta cgcctgaacc cagctttctt
     3241 gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     3301 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     3361 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3421 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3481 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3541 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3601 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3661 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3721 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3781 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3841 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3901 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3961 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4021 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4081 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4141 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4201 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4261 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4321 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4381 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4441 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4501 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4561 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4621 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4681 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4741 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4801 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4861 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4921 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4981 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5041 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5101 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5161 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5221 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5281 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5341 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5401 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5461 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5521 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5581 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5641 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5701 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5761 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5821 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5881 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5941 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6001 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6061 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6121 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6181 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6241 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6301 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6361 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6421 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6481 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6541 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6601 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6661 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6721 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6781 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6841 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6901 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6961 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7021 ccagtctatt
//
