LOCUS       sequence-236021-        9671 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 9671)
  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 9671)
  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: pHBS1400 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 G2...
FEATURES             Location/Qualifiers
     source          1..9671
                     /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(6242..6266)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7089..7144
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7542..7559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7713..8301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7793..7812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8472..9263,9264..9332))
                     /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     9095..9114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9333..9437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9505..9523
                     /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 tatgtctgaa tatattcggg taaccgaaga tgagaacgat gagcccattg
     5041 aaataccatc ggaagacgat gggacggtgc tgctctccac ggttacagcc cagtttccag
     5101 gggcgtgtgg gcttcgctac aggaatccag tgtctcagtg tatgagaggt gtccggctgg
     5161 tagaaggaat tctgcatgcc ccagatgctg gctggggaaa tctggtgtat gttgtcaact
     5221 atccaaaaga taacaaaaga aaaatggatg agacagatgc ttcatcagca gtgaaagtga
     5281 aaagagcagt ccagaaaaca tccgatttaa tagtgttggg tctcccatgg aaaacaaccg
     5341 aacaggacct gaaagagtat tttagtacct ttggagaagt tcttatggtg caggtcaaga
     5401 aagatcttaa gactggtcat tcaaaggggt ttggctttgt tcgttttacg gaatatgaaa
     5461 cacaagtgaa agtaatgtca cagcgacata tgatagatgg acgatggtgt gactgcaaac
     5521 ttcctaattc taagcaaagc caagatgagc ctttgagaag cagaaaagtg tttgtggggc
     5581 gctgtacaga ggacatgact gaggatgagc tgcgggagtt cttctctcag tacggggatg
     5641 tgatggatgt cttcatcccc aagccattca gggcctttgc ctttgttaca tttgcagatg
     5701 atcagattgc gcagtctctt tgtggagagg acttgatcat taaaggaatc agcgttcata
     5761 tatccaatgc cgaacctaag cacaatagca atagacagtt agaaagaagt ggaagatttg
     5821 gtggtaatcc aggtggcttt gggaatcagg gtggatttgg taatagcaga gggtttggag
     5881 ctggtttggg aaacaatcaa ggtagtaata tgggtggtgg gatgaacttt ggtgcgttca
     5941 gcattaatcc agccatgatg gctgccgccc aggcagcact acagagcagt tggggtatga
     6001 tgggcatgtt agccagccag cagaaccagt caggcccatc gggtaataac caaaaccaag
     6061 gcaacatgca gagggagcca aaccaggcct tcggttctgg aaataactct tatagtggct
     6121 ctaattctgg tgcagcaatt ggttggggat cagcatccaa tgcagggtcg ggcagtggtt
     6181 ttaatggagg ctttggctca agcatggatt ctaagtcttc tggctgggga atgtagacta
     6241 gacccagctt tcttgtacaa agtggttgat cctctagact gagaacttca gggtgagttt
     6301 ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggmgstat
     6361 tgtaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag
     6421 atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg
     6481 ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt
     6541 tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta
     6601 ctttctctaa tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca
     6661 gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc
     6721 tggctggcgt ggaaatattc ttattggtag aaacaactac accctggtca tcatcctgcc
     6781 tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc
     6841 caaaccgggc ccctctgcta accatgttca tgccttcttc tctttcctac agctcctggg
     6901 caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc ctcaggtgca
     6961 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     7021 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     7081 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     7141 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     7201 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     7261 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     7321 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     7381 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     7441 catagctgtc cctcttctct tatggagatc cctcgactgc attaatgaat cggccaacgc
     7501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9061 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     9121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9181 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9421 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     9481 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactcg
     9541 acgcggccag cttggtcgag ctggatactt cccgtccgcc agggggacat gccggcgatg
     9601 ctgaaggtcg cgcgcattcc cgatgaagag gccggttacc gcctgttgac ctggtgggac
     9661 gggcagggcg c
//
