LOCUS       sequence-229724-        9672 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 9672)
  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 9672)
  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: pHBS1396 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 WT]
FEATURES             Location/Qualifiers
     source          1..9672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..1372
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1373..1438
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1463..1519
                     /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(2770..2794)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2849..3421
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3618..3673
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4071..4088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4322..4341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5001..5792,5793..5861))
                     /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     5624..5643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5862..5966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6034..6052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6527..6582
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6779..7351
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7697..7716)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7966..7984
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9606..9615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cctcccaccg tacacgccac ctcgacatac tcgagtagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      421 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      481 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      541 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      601 gctggtttag tgaaccgtca gatccgctag ggatcctcta gtcagctgac gcgttaccac
      661 cggtgccacc atgggaagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
      721 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
      781 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
      841 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
      901 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
      961 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     1021 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     1081 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     1141 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac
     1201 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     1261 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     1321 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attccggact
     1381 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     1441 cggatctaga ggaggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt
     1501 ggaggagaac cctggaccta tgtctgaata tattcgggta accgaagatg agaacgatga
     1561 gcccattgaa ataccatcgg aagacgatgg gacggtgctg ctctccacgg ttacagccca
     1621 gtttccaggg gcgtgtgggc ttcgctacag gaatccagtg tctcagtgta tgagaggtgt
     1681 ccggctggta gaaggaattc tgcatgcccc agatgctggc tggggaaatc tggtgtatgt
     1741 tgtcaactat ccaaaagata acaaaagaaa aatggatgag acagatgctt catcagcagt
     1801 gaaagtgaaa agagcagtcc agaaaacatc cgatttaata gtgttgggtc tcccatggaa
     1861 aacaaccgaa caggacctga aagagtattt tagtaccttt ggagaagttc ttatggtgca
     1921 ggtcaagaaa gatcttaaga ctggtcattc aaaggggttt ggctttgttc gttttacgga
     1981 atatgaaaca caagtgaaag taatgtcaca gcgacatatg atagatggac gatggtgtga
     2041 ctgcaaactt cctaattcta agcaaagcca agatgagcct ttgagaagca gaaaagtgtt
     2101 tgtggggcgc tgtacagagg acatgactga ggatgagctg cgggagttct tctctcagta
     2161 cggggatgtg atggatgtct tcatccccaa gccattcagg gcctttgcct ttgttacatt
     2221 tgcagatgat cagattgcgc agtctctttg tggagaggac ttgatcatta aaggaatcag
     2281 cgttcatata tccaatgccg aacctaagca caatagcaat agacagttag aaagaagtgg
     2341 aagatttggt ggtaatccag gtggctttgg gaatcagggt ggatttggta atagcagagg
     2401 gggtggagct ggtttgggaa acaatcaagg tagtaatatg ggtggtggga tgaactttgg
     2461 tgcgttcagc attaatccag ccatgatggc tgccgcccag gcagcactac agagcagttg
     2521 gggtatgatg ggcatgttag ccagccagca gaaccagtca ggcccatcgg gtaataacca
     2581 aaaccaaggc aacatgcaga gggagccaaa ccaggccttc ggttctggaa ataactctta
     2641 tagtggctct aattctggtg cagcaattgg ttggggatca gcatccaatg cagggtcggg
     2701 cagtggtttt aatggaggct ttggctcaag catggattct aagtcttctg gctggggaat
     2761 gtagactaga cccagctttc ttgtacaaag tggttgatcc tctactagta agcttgcatg
     2821 cgatatctct agagctgaga acttcagggt gagtttgggg acccttgatt gttctttctt
     2881 tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg tgttgtttag
     2941 aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt tctttcactt
     3001 tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg taactttttc
     3061 gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt atttgtcaga
     3121 ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat tatattgtac
     3181 ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa tatttctgca
     3241 tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca ccctggtcat
     3301 catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga ggataaaata
     3361 ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct ctttcctaca
     3421 gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag aattcactcc
     3481 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     3541 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     3601 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3661 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     3721 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     3781 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     3841 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     3901 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     3961 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgactgca ttaatgaatc
     4021 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4681 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     5641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6001 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6061 cttcactcga cgcggccagc ttggtcgagc tggatacttc ccgtccgcca gggggacatg
     6121 ccggcgatgc tgaaggtcgc gcgcattccc gatgaagagg ccggttaccg cctgttgacc
     6181 tggtgggacg ggcagggcgc cgagttcata agagaagagg gacagctatg actgggagta
     6241 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     6301 aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc
     6361 tatggaataa ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc
     6421 aacctttgtt catggcagcc agcatatggg catatgttgc caaactctaa accaaatact
     6481 cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa
     6541 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
     6601 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct cagtggtatt
     6661 tgtgagccag ggcattggcc acaccagcca ccaccttctg ataggcagcc tgcacctgag
     6721 gagtgaattc tttgccaaaa tgatgagaca gcacaacaac cagcacgttg cccaggagct
     6781 gtaggaaaga gaagaaggca tgaacatggt tagcagaggg gcccggtttg gactcagagt
     6841 attttatcct catctcaaac agtgtatatc attgtaacca taaagagaaa ggcaggatga
     6901 tgaccagggt gtagttgttt ctaccaataa gaatatttcc acgccagcca gaatttatat
     6961 gcagaaatat tctaccttat catttaatta taacaattgt tctctaaaac tgtgctgaag
     7021 tacaatataa tataccctga ttgccttgaa aaaaaagtga ttagagaaag tacttacaat
     7081 ctgacaaata aacaaaagtg aatttaaaaa ttcgttacaa atgcaagcta aagtttaacg
     7141 aaaaagttac agaaaatgaa aagaaaataa gaggagacaa tggttgtcaa cagagtagaa
     7201 agtgaaagaa acaaaattat catgagggtc catggtgata caagggacat cttcccattc
     7261 taaacaacac cctgaaaact ttgccccctc catataacat gaattttaca atagcgaaaa
     7321 agaaagaaca atcaagggtc cccaaactca ccctgaagtt ctcagtctag agtcgagagg
     7381 atcaatcact ttgtacaaga aagctgggtc tagatagctt acttgtacag ctcgtccatg
     7441 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg
     7501 ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc
     7561 ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc
     7621 agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag
     7681 gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg
     7741 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc
     7801 accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     7861 agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg
     7921 aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc
     7981 ttggcggtct gggtgccctc gtaggggcgg ccctgaggaa aaaaaaggaa agggcatgct
     8041 cgccatgtgc aagatacagt gttgaatgtg gtgcaaaata taaagatgtg gaagaaaaag
     8101 ccactgaaaa taatatgagg aaataaaaaa ctagagtaaa ttatcagtgg aacaatttct
     8161 attatacaag acactacacc catacattct cctaatgctc atgtaagaat ttctccaaaa
     8221 ataccttcca gcactacaaa ctagaaaaaa aaagagacat ggacaccaaa ttaagttctt
     8281 aatagtgaag aacaaaagaa ctaccttgcc tgctccagtg gatccagcaa ccgccaacaa
     8341 ctgtcctctt tctatcttga aattaatatc tttcaggaca ggagtaccaa gaagtgagaa
     8401 attactgaag aagaggctgt catcaccatt agaagttttt ctattgttat tgttttgttt
     8461 tgctttctca aataattccc caaatccctg ttaaaaaaaa acacacacac acacacacac
     8521 acacacatca aaaataaaag atgagtttgt cagatatttt caatagatgc ttgttttatg
     8581 taccatactc acctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     8641 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
     8701 gctggttccg gtgcctccac tttcatcttc acctttagtg ataccggctg ccgtcacgaa
     8761 ctccttcagg accatgtgat cgcgcttctc gttcgggtct ttgctcagcg tggtctgggt
     8821 gctcagatag tggttgtccg gcaggcgcac cgggccgtcg ccgatcgggg tgttctgctg
     8881 tttgtggtcg gcttcctgct ggctgccgtc ctcaacgttg tggcggatct tgaagttagc
     8941 cttgatgccg ttcttctgct tgtcggcttc gatacggacg ttgtggctgt tgtggttgta
     9001 ctccagcttg tgacccagga tgttgccgtc ctccttgaag tcgttgccct tcagctcgat
     9061 gcggttcacc agggtgtcgc cctcgaactt cacctccgcg cgggtcttgt aggtgccgtc
     9121 gtccttgaac tcgatggtgc gctcctggac gtagccttcc ggcatggcgg acttgaagaa
     9181 gtcgtgacgc ttcatgtggt ccgggtagcg gctgaagcac tgcacgccgt aggtcagggt
     9241 ggtcacgagg gtcggccacg gcaccggcag cttgccggtg gtgcagatga acttcagggt
     9301 cagcttgccc tcggtggcat cgccctcgcc ctcgccacgc acgctgaact tgtggccgtt
     9361 tacgtcgccg tccagctcga ctttgatagg cacagtacct gtaaatagtt cctctccttt
     9421 actggtacca tcatcaggga atgagcttac atttctcctc ttcagcatct ggtcatcctt
     9481 cttagctttc ctcagctcca cattgacctc tattctgcga cgcctcattt ctgtactgtc
     9541 ttttcccttg ttcttgaatc tgtgaagacg ggcagctggt gtattagcat tctcgttggt
     9601 ggatcccatg gtggcaccgg tgaattctcc aggcgatctg acggttcact aaacgagctc
     9661 tgcttatata gg
//
