LOCUS       sequence-235268-        9640 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 9640)
  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 9640)
  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: pHBS1404 [IBB-GFP-mCherry 3E]-[BFP-P2A-TDP43 V...
FEATURES             Location/Qualifiers
     source          1..9640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        424..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     578..598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      689..698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             695..1396
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1397..1462
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1487..1543
                     /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(2789..2813)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2841..3413
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3610..3665
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4063..4080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4234..4822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4314..4333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4993..5784,5785..5853))
                     /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     5616..5635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5854..5958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6026..6044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6519..6574
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6771..7343
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7689..7708)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7958..7976
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9598..9607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctaaacgagc tctgcttata taggcctccc accgtacacg ccacctcgac atactcgagt
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagggatcc
      661 tctagtcagc tgacgcgtta ccaccggtgc caccatggga agcgagctga ttaaggagaa
      721 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc
      781 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
      841 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac
      901 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt
      961 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     1021 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc
     1081 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta
     1141 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag
     1201 ccatctgatc gcaaacgcca agaccacata tagatccaag aaacccgcta agaacctcaa
     1261 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga
     1321 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
     1381 ggggcacaag cttaattccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
     1441 cggtaccgcg ggcccgggat ccaccggatc tagaggagga agcggagcta ctaacttcag
     1501 cctgctgaag caggctggag acgtggagga gaaccctgga cctatgtctg aatatattcg
     1561 ggtaaccgaa gatgagaacg atgagcccat tgaaatacca tcggaagacg atgggacggt
     1621 gctgctctcc acggttacag cccagtttcc aggggcgtgt gggcttcgct acaggaatcc
     1681 agtgtctcag tgtatgagag gtgtccggct ggtagaagga attctgcatg ccccagatgc
     1741 tggctgggga aatctggtgt atgttgtcaa ctatccaaaa gataacaaaa gaaaaatgga
     1801 tgagacagat gcttcatcag cagtgaaagt gaaaagagca gtccagaaaa catccgattt
     1861 aatagtgttg ggtctcccat ggaaaacaac cgaacaggac ctgaaagagt attttagtac
     1921 ctttggagaa gttcttatgg tgcaggtcaa gaaagatctt aagactggtc attcaaaggg
     1981 gtttggcttt gttcgtttta cggaatatga aacacaagtg aaagtaatgt cacagcgaca
     2041 tatgatagat ggacgatggt gtgactgcaa acttcctaat tctaagcaaa gccaagatga
     2101 gcctttgaga agcagaaaag tgtttgtggg gcgctgtaca gaggacatga ctgaggatga
     2161 gctgcgggag ttcttctctc agtacgggga tgtgatggat gtcttcatcc ccaagccatt
     2221 cagggccttt gcctttgtta catttgcaga tgatcagatt gcgcagtctc tttgtggaga
     2281 ggacttgatc attaaaggaa tcagcgttca tatatccaat gccgaaccta agcacaacag
     2341 taatcgacag tccgagagaa gcgggcggtt cggtggtaat ccaggggggt tcggaaacca
     2401 aggaggtttt ggtaactcta ggggaggagg cgctggttcc ggtaataacc aaggcagcaa
     2461 cagtgggggg ggcagtaatt tcggtgcttt ttcaagtaac ccagcaatga tggccgccgc
     2521 acaagcagct ttgcaaagtt cttggggcat gatgggcatg ctggcgtcac agcaaaatca
     2581 aagcggaccc tcaggaaata atcaaaacca aggcaatagc cagagggagc ctaatcaggc
     2641 cttcggatca ggcaataatt catacagtgg ttcaaactcc ggggcagcct ctgggtgggg
     2701 cagcgcaagt aatgccggaa gcggtagtgg attcaacgga gggttcgggt caagtagcga
     2761 tagtaaaagt tccggttggg gctcttagac ccagctttct tgtacaaagt ggttgatcct
     2821 ctagagctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     2881 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     2941 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     3001 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     3061 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     3121 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     3181 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     3241 ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc
     3301 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     3361 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg
     3421 gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcact cctcaggtgc
     3481 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     3541 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     3601 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     3661 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     3721 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     3781 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     3841 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     3901 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     3961 tcatagctgt ccctcttctc ttatggagat ccctcgactg cattaatgaa tcggccaacg
     4021 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4081 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4141 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5581 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     5641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6001 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcactc
     6061 gacgcggcca gcttggtcga gctggatact tcccgtccgc cagggggaca tgccggcgat
     6121 gctgaaggtc gcgcgcattc ccgatgaaga ggccggttac cgcctgttga cctggtggga
     6181 cgggcagggc gccgagttca taagagaaga gggacagcta tgactgggag tagtcaggag
     6241 aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa
     6301 aataacacaa aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat
     6361 aaggaatgga cagcaggggg ctgtttcata tactgatgac ctctttatag ccaacctttg
     6421 ttcatggcag ccagcatatg ggcatatgtt gccaaactct aaaccaaata ctcattctga
     6481 tgttttaaat gatttgccct cccatatgtc cttccgagtg agagacacaa aaaattccaa
     6541 cacactattg caatgaaaat aaatttcctt tattagccag aagtcagatg ctcaaggggc
     6601 ttcatgatgt ccccataatt tttggcagag ggaaaaagat ctcagtggta tttgtgagcc
     6661 agggcattgg ccacaccagc caccaccttc tgataggcag cctgcacctg aggagtgaat
     6721 tctttgccaa aatgatgaga cagcacaaca accagcacgt tgcccaggag ctgtaggaaa
     6781 gagaagaagg catgaacatg gttagcagag gggcccggtt tggactcaga gtattttatc
     6841 ctcatctcaa acagtgtata tcattgtaac cataaagaga aaggcaggat gatgaccagg
     6901 gtgtagttgt ttctaccaat aagaatattt ccacgccagc cagaatttat atgcagaaat
     6961 attctacctt atcatttaat tataacaatt gttctctaaa actgtgctga agtacaatat
     7021 aatataccct gattgccttg aaaaaaaagt gattagagaa agtacttaca atctgacaaa
     7081 taaacaaaag tgaatttaaa aattcgttac aaatgcaagc taaagtttaa cgaaaaagtt
     7141 acagaaaatg aaaagaaaat aagaggagac aatggttgtc aacagagtag aaagtgaaag
     7201 aaacaaaatt atcatgaggg tccatggtga tacaagggac atcttcccat tctaaacaac
     7261 accctgaaaa ctttgccccc tccatataac atgaatttta caatagcgaa aaagaaagaa
     7321 caatcaaggg tccccaaact caccctgaag ttctcagtct agagtcgaga ggatcaatca
     7381 ctttgtacaa gaaagctggg tctagatagc ttacttgtac agctcgtcca tgccgccggt
     7441 ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct cgttgtggga
     7501 ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc
     7561 cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct tcagcctctg
     7621 cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca
     7681 gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt
     7741 caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc
     7801 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa
     7861 gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg
     7921 ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca gcttggcggt
     7981 ctgggtgccc tcgtaggggc ggccctgagg aaaaaaaagg aaagggcatg ctcgccatgt
     8041 gcaagataca gtgttgaatg tggtgcaaaa tataaagatg tggaagaaaa agccactgaa
     8101 aataatatga ggaaataaaa aactagagta aattatcagt ggaacaattt ctattataca
     8161 agacactaca cccatacatt ctcctaatgc tcatgtaaga atttctccaa aaataccttc
     8221 cagcactaca aactagaaaa aaaaagagac atggacacca aattaagttc ttaatagtga
     8281 agaacaaaag aactaccttg cctgctccag tggatccagc aaccgccaac aactgtcctc
     8341 tttctatctt gaaattaata tctttcagga caggagtacc aagaagtgag aaattactga
     8401 agaagaggct gtcatcacca ttagaagttt ttctattgtt attgttttgt tttgctttct
     8461 caaataattc cccaaatccc tgttaaaaaa aaacacacac acacacacac acacacacat
     8521 caaaaataaa agatgagttt gtcagatatt ttcaatagat gcttgtttta tgtaccatac
     8581 tcacctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc tccatgtgca
     8641 ccttgaagcg catgaactcc ttgatgatgg ccatgttatc ctcctcgccc ttgctggttc
     8701 cggtgcctcc actttcatct tcacctttag tgataccggc tgccgtcacg aactccttca
     8761 ggaccatgtg atcgcgcttc tcgttcgggt ctttgctcag cgtggtctgg gtgctcagat
     8821 agtggttgtc cggcaggcgc accgggccgt cgccgatcgg ggtgttctgc tgtttgtggt
     8881 cggcttcctg ctggctgccg tcctcaacgt tgtggcggat cttgaagtta gccttgatgc
     8941 cgttcttctg cttgtcggct tcgatacgga cgttgtggct gttgtggttg tactccagct
     9001 tgtgacccag gatgttgccg tcctccttga agtcgttgcc cttcagctcg atgcggttca
     9061 ccagggtgtc gccctcgaac ttcacctccg cgcgggtctt gtaggtgccg tcgtccttga
     9121 actcgatggt gcgctcctgg acgtagcctt ccggcatggc ggacttgaag aagtcgtgac
     9181 gcttcatgtg gtccgggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     9241 gggtcggcca cggcaccggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     9301 cctcggtggc atcgccctcg ccctcgccac gcacgctgaa cttgtggccg tttacgtcgc
     9361 cgtccagctc gactttgata ggcacagtac ctgtaaatag ttcctctcct ttactggtac
     9421 catcatcagg gaatgagctt acatttctcc tcttcagcat ctggtcatcc ttcttagctt
     9481 tcctcagctc cacattgacc tctattctgc gacgcctcat ttctgtactg tcttttccct
     9541 tgttcttgaa tctgtgaaga cgggcagctg gtgtattagc attctcgttg gtggatccca
     9601 tggtggcacc ggtgaattct ccaggcgatc tgacggttca
//
