LOCUS       sequence-262084-        9499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 splicing activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9499)
  AUTHORS   Schmidt HB, Barreau A, Rohatgi R
  TITLE     Phase separation-deficient TDP43 remains functional in splicing.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
  PUBMED    31653829
REFERENCE   2  (bases 1 to 9499)
  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: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: " 
            10.1038/s41467-019-12740-2"
COMMENT     Sequence Label: pHBS1503 [IBB-GFP-mCherry3E]-[BFP-TDP43 WT]
FEATURES             Location/Qualifiers
     source          1..9499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    508..532
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1784..2485)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     regulatory      2482..2491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2553..2756)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2582..2602)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2757..3060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3213..3222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4844..4862)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5112..5131
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          5477..6049
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6246..6301
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6699..6716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6870..7458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6950..6969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7629..8420,8421..8489))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8252..8271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8490..8594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8662..8680
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    9155..9210
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          join(9407..9499,1..480)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 ttgtaaccat aaagagaaag gcaggatgat gaccagggtg tagttgtttc taccaataag
       61 aatatttcca cgccagccag aatttatatg cagaaatatt ctaccttatc atttaattat
      121 aacaattgtt ctctaaaact gtgctgaagt acaatataat ataccctgat tgccttgaaa
      181 aaaaagtgat tagagaaagt acttacaatc tgacaaataa acaaaagtga atttaaaaat
      241 tcgttacaaa tgcaagctaa agtttaacga aaaagttaca gaaaatgaaa agaaaataag
      301 aggagacaat ggttgtcaac agagtagaaa gtgaaagaaa caaaattatc atgagggtcc
      361 atggtgatac aagggacatc ttcccattct aaacaacacc ctgaaaactt tgccccctcc
      421 atataacatg aattttacaa tagcgaaaaa gaaagaacaa tcaagggtcc ccaaactcac
      481 cctgaagttc tcagctctag aggatcaacc actttgtaca agaaagctgg gtctacattc
      541 cccagccaga agacttagaa tccatgcttg agccaaagcc tccattaaaa ccactgcccg
      601 accctgcatt ggatgctgat ccccaaccaa ttgctgcacc agaattagag ccactataag
      661 agttatttcc agaaccgaag gcctggtttg gctccctctg catgttgcct tggttttggt
      721 tattacccga tgggcctgac tggttctgct ggctggctaa catgcccatc ataccccaac
      781 tgctctgtag tgctgcctgg gcggcagcca tcatggctgg attaatgctg aacgcaccaa
      841 agttcatccc accacccata ttactacctt gattgtttcc caaaccagct ccaccccctc
      901 tgctattacc aaatccaccc tgattcccaa agccacctgg attaccacca aatcttccac
      961 ttctttctaa ctgtctattg ctattgtgct taggttcggc attggatata tgaacgctga
     1021 ttcctttaat gatcaagtcc tctccacaaa gagactgcgc aatctgatca tctgcaaatg
     1081 taacaaaggc aaaggccctg aatggcttgg ggatgaagac atccatcaca tccccgtact
     1141 gagagaagaa ctcccgcagc tcatcctcag tcatgtcctc tgtacagcgc cccacaaaca
     1201 cttttctgct tctcaaaggc tcatcttggc tttgcttaga attaggaagt ttgcagtcac
     1261 accatcgtcc atctatcata tgtcgctgtg acattacttt cacttgtgtt tcatattccg
     1321 taaaacgaac aaagccaaac ccctttgaat gaccagtctt aagatctttc ttgacctgca
     1381 ccataagaac ttctccaaag gtactaaaat actctttcag gtcctgttcg gttgttttcc
     1441 atgggagacc caacactatt aaatcggatg ttttctggac tgctcttttc actttcactg
     1501 ctgatgaagc atctgtctca tccatttttc ttttgttatc ttttggatag ttgacaacat
     1561 acaccagatt tccccagcca gcatctgggg catgcagaat tccttctacc agccggacac
     1621 ctctcataca ctgagacact ggattcctgt agcgaagccc acacgcccct ggaaactggg
     1681 ctgtaaccgt ggagagcagc accgtcccat cgtcttccga tggtatttca atgggctcat
     1741 cgttctcatc ttcggttacc cgaatatatt cagacattcc ggaattaagc ttgtgcccca
     1801 gtttgctagg gaggtcgcag tatctggcca ctgccacctc gtgctgctcg acgtaggtct
     1861 cgttgttggc ctccttgatt ctttccagtc tgtagtccac atagtagacg ccaggcatct
     1921 tgaggttctt agcgggtttc ttggatctat atgtggtctt ggcgtttgcg atcagatggc
     1981 tcccgcccac gagcttcagg gccatgtcgt ttctgccttc caggccgccg tcagcggggt
     2041 acagcgtctc ggtgaaggcc tcccagccga gtgttttctt ctgcatcaca gggccgttgg
     2101 atgtgaagtt cacccctctg atcttgacgt tgtagatgag gcagccgtcc tggaggctgg
     2161 tgtcctgggt agcggtcagc acgcccccgt cttcgtatgt ggtgactctc tcccatgtga
     2221 agccctcagg gaaggactgc ttgaagaagt cggggatgcc ctgggtgtgg ttgatgaagg
     2281 tcttgctgcc gtagaggaag ctagtagcca ggatgtcgaa ggcgaagggg agagggccgc
     2341 cctcgaccac cttgattctc atggtctggg tgccctcgta gggcttgcct tcgccctcgg
     2401 atgtgcactt gaagtgatgg ttgtccacgg tgccctccat gtacagcttc atgtgcatgt
     2461 tctccttaat cagctcgctt cccatggtgg caccggtggt aacgcgtcag ctgactagag
     2521 gatccctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt
     2581 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     2641 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     2701 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     2761 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     2821 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     2881 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     2941 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     3001 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     3061 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     3121 ctcgagtatg tcgaggtggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt
     3181 gaaccgtcag atcgcctgga gaattcaccg gtgccaccat gggatccacc aacgagaatg
     3241 ctaatacacc agctgcccgt cttcacagat tcaagaacaa gggaaaagac agtacagaaa
     3301 tgaggcgtcg cagaatagag gtcaatgtgg agctgaggaa agctaagaag gatgaccaga
     3361 tgctgaagag gagaaatgta agctcattcc ctgatgatgg taccagtaaa ggagaggaac
     3421 tatttacagg tactgtgcct atcaaagtcg agctggacgg cgacgtaaac ggccacaagt
     3481 tcagcgtgcg tggcgagggc gagggcgatg ccaccgaggg caagctgacc ctgaagttca
     3541 tctgcaccac cggcaagctg ccggtgccgt ggccgaccct cgtgaccacc ctgacctacg
     3601 gcgtgcagtg cttcagccgc tacccggacc acatgaagcg tcacgacttc ttcaagtccg
     3661 ccatgccgga aggctacgtc caggagcgca ccatcgagtt caaggacgac ggcacctaca
     3721 agacccgcgc ggaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3781 gcaacgactt caaggaggac ggcaacatcc tgggtcacaa gctggagtac aaccacaaca
     3841 gccacaacgt ccgtatcgaa gccgacaagc agaagaacgg catcaaggct aacttcaaga
     3901 tccgccacaa cgttgaggac ggcagccagc aggaagccga ccacaaacag cagaacaccc
     3961 cgatcggcga cggcccggtg cgcctgccgg acaaccacta tctgagcacc cagaccacgc
     4021 tgagcaaaga cccgaacgag aagcgcgatc acatggtcct gaaggagttc gtgacggcag
     4081 ccggtatcac taaaggtgaa gatgaaagtg gaggcaccgg aaccagcaag ggcgaggagg
     4141 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     4201 acggccacga gttcgagatc gagggcgagg gcgaggtgag tatggtacat aaaacaagca
     4261 tctattgaaa atatctgaca aactcatctt ttatttttga tgtgtgtgtg tgtgtgtgtg
     4321 tgtgtgtttt tttttaacag ggatttgggg aattatttga gaaagcaaaa caaaacaata
     4381 acaatagaaa aacttctaat ggtgatgaca gcctcttctt cagtaatttc tcacttcttg
     4441 gtactcctgt cctgaaagat attaatttca agatagaaag aggacagttg ttggcggttg
     4501 ctggatccac tggagcaggc aaggtagttc ttttgttctt cactattaag aacttaattt
     4561 ggtgtccatg tctctttttt tttctagttt gtagtgctgg aaggtatttt tggagaaatt
     4621 cttacatgag cattaggaga atgtatgggt gtagtgtctt gtataataga aattgttcca
     4681 ctgataattt actctagttt tttatttcct catattattt tcagtggctt tttcttccac
     4741 atctttatat tttgcaccac attcaacact gtatcttgca catggcgagc atgccctttc
     4801 cttttttttc ctcagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     4861 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     4921 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     4981 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     5041 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     5101 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     5161 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     5221 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     5281 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     5341 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     5401 tacaagtaag ctatctagac ccagctttct tgtacaaagt gattgatcct ctcgactcta
     5461 gactgagaac ttcagggtga gtttggggac ccttgattgt tctttctttt tcgctattgt
     5521 aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg
     5581 tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc tactctgttg
     5641 acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt taaactttag
     5701 cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt gtaagtactt
     5761 tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt cagcacagtt
     5821 ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata taaattctgg
     5881 ctggcgtgga aatattctta ttggtagaaa caactacacc ctggtcatca tcctgccttt
     5941 ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact ctgagtccaa
     6001 accgggcccc tctgctaacc atgttcatgc cttcttctct ttcctacagc tcctgggcaa
     6061 cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttcactcctc aggtgcaggc
     6121 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     6181 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     6241 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     6301 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     6361 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     6421 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     6481 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     6541 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     6601 agctgtccct cttctcttat ggagatccct cgactgcatt aatgaatcgg ccaacgcgcg
     6661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8221 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     8281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8641 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcactcgacg
     8701 cggccagctt ggtcgagctg gatacttccc gtccgccagg gggacatgcc ggcgatgctg
     8761 aaggtcgcgc gcattcccga tgaagaggcc ggttaccgcc tgttgacctg gtgggacggg
     8821 cagggcgccg agttcataag agaagaggga cagctatgac tgggagtagt caggagagga
     8881 ggaaaaatct ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata
     8941 acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa ggcttttcta tggaataagg
     9001 aatggacagc agggggctgt ttcatatact gatgacctct ttatagccaa cctttgttca
     9061 tggcagccag catatgggca tatgttgcca aactctaaac caaatactca ttctgatgtt
     9121 ttaaatgatt tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttccaacaca
     9181 ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca
     9241 tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg
     9301 cattggccac accagccacc accttctgat aggcagcctg cacctgagga gtgaattctt
     9361 tgccaaaatg atgagacagc acaacaacca gcacgttgcc caggagctgt aggaaagaga
     9421 agaaggcatg aacatggtta gcagaggggc ccggtttgga ctcagagtat tttatcctca
     9481 tctcaaacag tgtatatca
//
