LOCUS       sequence-229887-        9369 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 9369)
  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 9369)
  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: pHBS1392 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43
            de...
FEATURES             Location/Qualifiers
     source          1..9369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    327..382
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          579..1151
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1497..1516)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1766..1784
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3406..3415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        3568..3871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3872..4075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4026..4046
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4137..4146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4143..4844
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    4845..4910
                     /label=MCS
                     /note="multiple cloning site"
     CDS             4935..4991
                     /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(5939..5963)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          6018..6590
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6787..6842
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7240..7257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7411..7999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7491..7510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8170..8961,8962..9030))
                     /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     8793..8812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9031..9135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9203..9221
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgagttcata agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat
       61 ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa
      121 caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca
      181 gcagggggct gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc
      241 agcatatggg catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga
      301 tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca
      361 atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc
      421 ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc
      481 acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaattc tttgccaaaa
      541 tgatgagaca gcacaacaac cagcacgttg cccaggagct gtaggaaaga gaagaaggca
      601 tgaacatggt tagcagaggg gcccggtttg gactcagagt attttatcct catctcaaac
      661 agtgtatatc attgtaacca taaagagaaa ggcaggatga tgaccagggt gtagttgttt
      721 ctaccaataa gaatatttcc acgccagcca gaatttatat gcagaaatat tctaccttat
      781 catttaatta taacaattgt tctctaaaac tgtgctgaag tacaatataa tataccctga
      841 ttgccttgaa aaaaaagtga ttagagaaag tacttacaat ctgacaaata aacaaaagtg
      901 aatttaaaaa ttcgttacaa atgcaagcta aagtttaacg aaaaagttac agaaaatgaa
      961 aagaaaataa gaggagacaa tggttgtcaa cagagtagaa agtgaaagaa acaaaattat
     1021 catgagggtc catggtgata caagggacat cttcccattc taaacaacac cctgaaaact
     1081 ttgccccctc catataacat gaattttaca atagcgaaaa agaaagaaca atcaagggtc
     1141 cccaaactca ccctgaagtt ctcagtctag agtcgagagg atcaatcact ttgtacaaga
     1201 aagctgggtc tagatagctt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     1261 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
     1321 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     1381 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     1441 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     1501 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     1561 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     1621 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     1681 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     1741 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     1801 gtaggggcgg ccctgaggaa aaaaaaggaa agggcatgct cgccatgtgc aagatacagt
     1861 gttgaatgtg gtgcaaaata taaagatgtg gaagaaaaag ccactgaaaa taatatgagg
     1921 aaataaaaaa ctagagtaaa ttatcagtgg aacaatttct attatacaag acactacacc
     1981 catacattct cctaatgctc atgtaagaat ttctccaaaa ataccttcca gcactacaaa
     2041 ctagaaaaaa aaagagacat ggacaccaaa ttaagttctt aatagtgaag aacaaaagaa
     2101 ctaccttgcc tgctccagtg gatccagcaa ccgccaacaa ctgtcctctt tctatcttga
     2161 aattaatatc tttcaggaca ggagtaccaa gaagtgagaa attactgaag aagaggctgt
     2221 catcaccatt agaagttttt ctattgttat tgttttgttt tgctttctca aataattccc
     2281 caaatccctg ttaaaaaaaa acacacacac acacacacac acacacatca aaaataaaag
     2341 atgagtttgt cagatatttt caatagatgc ttgttttatg taccatactc acctcgccct
     2401 cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca
     2461 tgaactcctt gatgatggcc atgttatcct cctcgccctt gctggttccg gtgcctccac
     2521 tttcatcttc acctttagtg ataccggctg ccgtcacgaa ctccttcagg accatgtgat
     2581 cgcgcttctc gttcgggtct ttgctcagcg tggtctgggt gctcagatag tggttgtccg
     2641 gcaggcgcac cgggccgtcg ccgatcgggg tgttctgctg tttgtggtcg gcttcctgct
     2701 ggctgccgtc ctcaacgttg tggcggatct tgaagttagc cttgatgccg ttcttctgct
     2761 tgtcggcttc gatacggacg ttgtggctgt tgtggttgta ctccagcttg tgacccagga
     2821 tgttgccgtc ctccttgaag tcgttgccct tcagctcgat gcggttcacc agggtgtcgc
     2881 cctcgaactt cacctccgcg cgggtcttgt aggtgccgtc gtccttgaac tcgatggtgc
     2941 gctcctggac gtagccttcc ggcatggcgg acttgaagaa gtcgtgacgc ttcatgtggt
     3001 ccgggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtcggccacg
     3061 gcaccggcag cttgccggtg gtgcagatga acttcagggt cagcttgccc tcggtggcat
     3121 cgccctcgcc ctcgccacgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3181 ctttgatagg cacagtacct gtaaatagtt cctctccttt actggtacca tcatcaggga
     3241 atgagcttac atttctcctc ttcagcatct ggtcatcctt cttagctttc ctcagctcca
     3301 cattgacctc tattctgcga cgcctcattt ctgtactgtc ttttcccttg ttcttgaatc
     3361 tgtgaagacg ggcagctggt gtattagcat tctcgttggt ggatcccatg gtggcaccgg
     3421 tgaattctcc aggcgatctg acggttcact aaacgagctc tgcttatata ggcctcccac
     3481 cgtacacgcc acctcgacat actcgagtag ttattaatag taatcaatta cggggtcatt
     3541 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3721 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3781 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3841 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3901 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3961 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4021 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     4081 agtgaaccgt cagatccgct agggatcctc tagtcagctg acgcgttacc accggtgcca
     4141 ccatgggaag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     4201 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     4261 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     4321 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     4381 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     4441 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     4501 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     4561 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
     4621 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
     4681 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     4741 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     4801 ccagatactg cgacctccct agcaaactgg ggcacaagct taattccgga ctcagatctc
     4861 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     4921 gaggaggaag cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga
     4981 accctggacc tatgacatcc gatttaatag tgttgggtct cccatggaaa acaaccgaac
     5041 aggacctgaa agagtatttt agtacctttg gagaagttct tatggtgcag gtcaagaaag
     5101 atcttaagac tggtcattca aaggggtttg gctttgttcg ttttacggaa tatgaaacac
     5161 aagtgaaagt aatgtcacag cgacatatga tagatggacg atggtgtgac tgcaaacttc
     5221 ctaattctaa gcaaagccaa gatgagcctt tgagaagcag aaaagtgttt gtggggcgct
     5281 gtacagagga catgactgag gatgagctgc gggagttctt ctctcagtac ggggatgtga
     5341 tggatgtctt catccccaag ccattcaggg cctttgcctt tgttacattt gcagatgatc
     5401 agattgcgca gtctctttgt ggagaggact tgatcattaa aggaatcagc gttcatatat
     5461 ccaatgccga acctaagcac aatagcaata gacagttaga aagaagtgga agatttggtg
     5521 gtaatccagg tggctttggg aatcagggtg gatttggtaa tagcagaggg ggtggagctg
     5581 gtttgggaaa caatcaaggt agtaatatgg gtggtgggat gaactttggt gcgttcagca
     5641 ttaatccagc catgatggct gccgcccagg cagcactaca gagcagttgg ggtatgatgg
     5701 gcatgttagc cagccagcag aaccagtcag gcccatcggg taataaccaa aaccaaggca
     5761 acatgcagag ggagccaaac caggccttcg gttctggaaa taactcttat agtggctcta
     5821 attctggtgc agcaattggt tggggatcag catccaatgc agggtcgggc agtggtttta
     5881 atggaggctt tggctcaagc atggattcta agtcttctgg ctggggaatg tagactagac
     5941 ccagctttct tgtacaaagt ggttgatcct ctactagtaa gcttgcatgc gatatctcta
     6001 gagctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     6061 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     6121 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     6181 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     6241 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     6301 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     6361 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     6421 gctggcgtgg aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt
     6481 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     6541 aaccgggccc ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca
     6601 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcactcct caggtgcagg
     6661 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     6721 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     6781 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     6841 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     6901 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     6961 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     7021 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     7081 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     7141 tagctgtccc tcttctctta tggagatccc tcgactgcat taatgaatcg gccaacgcgc
     7201 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7261 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7321 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7381 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7921 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7981 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8041 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8101 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8161 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8221 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8281 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8341 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8401 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8461 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8521 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8581 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8641 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8701 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8761 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     8821 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8881 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8941 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9001 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9121 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     9181 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcactcgac
     9241 gcggccagct tggtcgagct ggatacttcc cgtccgccag ggggacatgc cggcgatgct
     9301 gaaggtcgcg cgcattcccg atgaagaggc cggttaccgc ctgttgacct ggtgggacgg
     9361 gcagggcgc
//
