LOCUS       sequence-231650-        9213 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 9213)
  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 9213)
  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: pHBS1393 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43
            de...
FEATURES             Location/Qualifiers
     source          1..9213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             690..1391
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1392..1457
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1482..1538
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(2330..2354)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2409..2981
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3178..3233
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3802..4390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4561..5352,5353..5421))
                     /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     5184..5203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5422..5526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5594..5612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6087..6142
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6339..6911
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7257..7276)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7526..7544
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9166..9175
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgagctctgc ttatataggc ctcccaccgt acacgccacc tcgacatact cgagtagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagg gatcctctag
      661 tcagctgacg cgttaccacc ggtgccacca tgggaagcga gctgattaag gagaacatgc
      721 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
      781 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
      841 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
      901 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
      961 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     1021 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     1081 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     1141 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     1201 tgatcgcaaa cgccaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     1261 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     1321 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     1381 acaagcttaa ttccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     1441 ccgcgggccc gggatccacc ggatctagag gaggaagcgg agctactaac ttcagcctgc
     1501 tgaagcaggc tggagacgtg gaggagaacc ctggacctat gtctgaatat attcgggtaa
     1561 ccgaagatga gaacgatgag cccattgaaa taccatcgga agacgatggg acggtgctgc
     1621 tctccacggt tacagcccag tttccagggg cgtgtgggct tcgctacagg aatccagtgt
     1681 ctcagtgtat gagaggtgtc cggctggtag aaggaattct gcatgcccca gatgctggct
     1741 ggggaaatct ggtgtatgtt gtcaactatc caaaagataa caaaagaaaa atggatgaga
     1801 cagatgcttc atcagcagtg aaagtgaaaa gagcagtcca gaaaacatcc tccaatgccg
     1861 aacctaagca caatagcaat agacagttag aaagaagtgg aagatttggt ggtaatccag
     1921 gtggctttgg gaatcagggt ggatttggta atagcagagg gggtggagct ggtttgggaa
     1981 acaatcaagg tagtaatatg ggtggtggga tgaactttgg tgcgttcagc attaatccag
     2041 ccatgatggc tgccgcccag gcagcactac agagcagttg gggtatgatg ggcatgttag
     2101 ccagccagca gaaccagtca ggcccatcgg gtaataacca aaaccaaggc aacatgcaga
     2161 gggagccaaa ccaggccttc ggttctggaa ataactctta tagtggctct aattctggtg
     2221 cagcaattgg ttggggatca gcatccaatg cagggtcggg cagtggtttt aatggaggct
     2281 ttggctcaag catggattct aagtcttctg gctggggaat gtagactaga cccagctttc
     2341 ttgtacaaag tggttgatcc tctactagta agcttgcatg cgatatctct agagctgaga
     2401 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     2461 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     2521 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     2581 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     2641 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     2701 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     2761 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     2821 gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct ttctctttat
     2881 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     2941 cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc aacgtgctgg
     3001 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     3061 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     3121 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     3181 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3241 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3301 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3361 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3421 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3481 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3541 ctcttctctt atggagatcc ctcgactgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5581 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcactcga cgcggccagc
     5641 ttggtcgagc tggatacttc ccgtccgcca gggggacatg ccggcgatgc tgaaggtcgc
     5701 gcgcattccc gatgaagagg ccggttaccg cctgttgacc tggtgggacg ggcagggcgc
     5761 cgagttcata agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat
     5821 ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa
     5881 caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca
     5941 gcagggggct gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc
     6001 agcatatggg catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga
     6061 tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca
     6121 atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc
     6181 ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc
     6241 acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaattc tttgccaaaa
     6301 tgatgagaca gcacaacaac cagcacgttg cccaggagct gtaggaaaga gaagaaggca
     6361 tgaacatggt tagcagaggg gcccggtttg gactcagagt attttatcct catctcaaac
     6421 agtgtatatc attgtaacca taaagagaaa ggcaggatga tgaccagggt gtagttgttt
     6481 ctaccaataa gaatatttcc acgccagcca gaatttatat gcagaaatat tctaccttat
     6541 catttaatta taacaattgt tctctaaaac tgtgctgaag tacaatataa tataccctga
     6601 ttgccttgaa aaaaaagtga ttagagaaag tacttacaat ctgacaaata aacaaaagtg
     6661 aatttaaaaa ttcgttacaa atgcaagcta aagtttaacg aaaaagttac agaaaatgaa
     6721 aagaaaataa gaggagacaa tggttgtcaa cagagtagaa agtgaaagaa acaaaattat
     6781 catgagggtc catggtgata caagggacat cttcccattc taaacaacac cctgaaaact
     6841 ttgccccctc catataacat gaattttaca atagcgaaaa agaaagaaca atcaagggtc
     6901 cccaaactca ccctgaagtt ctcagtctag agtcgagagg atcaatcact ttgtacaaga
     6961 aagctgggtc tagatagctt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     7021 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
     7081 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     7141 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     7201 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     7261 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     7321 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     7381 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     7441 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     7501 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     7561 gtaggggcgg ccctgaggaa aaaaaaggaa agggcatgct cgccatgtgc aagatacagt
     7621 gttgaatgtg gtgcaaaata taaagatgtg gaagaaaaag ccactgaaaa taatatgagg
     7681 aaataaaaaa ctagagtaaa ttatcagtgg aacaatttct attatacaag acactacacc
     7741 catacattct cctaatgctc atgtaagaat ttctccaaaa ataccttcca gcactacaaa
     7801 ctagaaaaaa aaagagacat ggacaccaaa ttaagttctt aatagtgaag aacaaaagaa
     7861 ctaccttgcc tgctccagtg gatccagcaa ccgccaacaa ctgtcctctt tctatcttga
     7921 aattaatatc tttcaggaca ggagtaccaa gaagtgagaa attactgaag aagaggctgt
     7981 catcaccatt agaagttttt ctattgttat tgttttgttt tgctttctca aataattccc
     8041 caaatccctg ttaaaaaaaa acacacacac acacacacac acacacatca aaaataaaag
     8101 atgagtttgt cagatatttt caatagatgc ttgttttatg taccatactc acctcgccct
     8161 cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca
     8221 tgaactcctt gatgatggcc atgttatcct cctcgccctt gctggttccg gtgcctccac
     8281 tttcatcttc acctttagtg ataccggctg ccgtcacgaa ctccttcagg accatgtgat
     8341 cgcgcttctc gttcgggtct ttgctcagcg tggtctgggt gctcagatag tggttgtccg
     8401 gcaggcgcac cgggccgtcg ccgatcgggg tgttctgctg tttgtggtcg gcttcctgct
     8461 ggctgccgtc ctcaacgttg tggcggatct tgaagttagc cttgatgccg ttcttctgct
     8521 tgtcggcttc gatacggacg ttgtggctgt tgtggttgta ctccagcttg tgacccagga
     8581 tgttgccgtc ctccttgaag tcgttgccct tcagctcgat gcggttcacc agggtgtcgc
     8641 cctcgaactt cacctccgcg cgggtcttgt aggtgccgtc gtccttgaac tcgatggtgc
     8701 gctcctggac gtagccttcc ggcatggcgg acttgaagaa gtcgtgacgc ttcatgtggt
     8761 ccgggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtcggccacg
     8821 gcaccggcag cttgccggtg gtgcagatga acttcagggt cagcttgccc tcggtggcat
     8881 cgccctcgcc ctcgccacgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     8941 ctttgatagg cacagtacct gtaaatagtt cctctccttt actggtacca tcatcaggga
     9001 atgagcttac atttctcctc ttcagcatct ggtcatcctt cttagctttc ctcagctcca
     9061 cattgacctc tattctgcga cgcctcattt ctgtactgtc ttttcccttg ttcttgaatc
     9121 tgtgaagacg ggcagctggt gtattagcat tctcgttggt ggatcccatg gtggcaccgg
     9181 tgaattctcc aggcgatctg acggttcact aaa
//
