LOCUS       sequence-231277-        9458 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 9458)
  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 9458)
  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: pHBS1397 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 W3...
FEATURES             Location/Qualifiers
     source          1..9458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             641..1342
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1343..1408
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1433..1489
                     /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(2735..2759)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2814..3386
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3827..3844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4757..5548,5549..5617))
                     /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     5380..5399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5618..5722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5790..5808
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6283..6338
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6384..6403
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          6535..7107
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7453..7472)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7722..7740
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9362..9371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcgagtagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 ggatcctcta gtcagctgac gcgttaccac cggtgccacc atgggaagcg agctgattaa
      661 ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg
      721 cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt
      781 cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag
      841 caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga
      901 gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca
      961 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
     1021 cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac
     1081 gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg
     1141 cgggagccat ctgatcgcaa acgccaagac cacatataga tccaagaaac ccgctaagaa
     1201 cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa
     1261 caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag
     1321 caaactgggg cacaagctta attccggact cagatctcga gctcaagctt cgaattctgc
     1381 agtcgacggt accgcgggcc cgggatccac cggatctaga ggaggaagcg gagctactaa
     1441 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta tgtctgaata
     1501 tattcgggta accgaagatg agaacgatga gcccattgaa ataccatcgg aagacgatgg
     1561 gacggtgctg ctctccacgg ttacagccca gtttccaggg gcgtgtgggc ttcgctacag
     1621 gaatccagtg tctcagtgta tgagaggtgt ccggctggta gaaggaattc tgcatgcccc
     1681 agatgctggc tggggaaatc tggtgtatgt tgtcaactat ccaaaagata acaaaagaaa
     1741 aatggatgag acagatgctt catcagcagt gaaagtgaaa agagcagtcc agaaaacatc
     1801 cgatttaata gtgttgggtc tcccatggaa aacaaccgaa caggacctga aagagtattt
     1861 tagtaccttt ggagaagttc ttatggtgca ggtcaagaaa gatcttaaga ctggtcattc
     1921 aaaggggttt ggctttgttc gttttacgga atatgaaaca caagtgaaag taatgtcaca
     1981 gcgacatatg atagatggac gatggtgtga ctgcaaactt cctaattcta agcaaagcca
     2041 agatgagcct ttgagaagca gaaaagtgtt tgtggggcgc tgtacagagg acatgactga
     2101 ggatgagctg cgggagttct tctctcagta cggggatgtg atggatgtct tcatccccaa
     2161 gccattcagg gcctttgcct ttgttacatt tgcagatgat cagattgcgc agtctctttg
     2221 tggagaggac ttgatcatta aaggaatcag cgttcatata tccaatgccg aacctaagca
     2281 caatagcaat agacagttag aaagaagtgg aagatttggt ggtaatccag gtggctttgg
     2341 gaatcagggt ggatttggta atagcagagg gggtggagct ggtttgggaa acaatcaagg
     2401 tagtaatatg ggtggtggga tgaactttgg tgcgttcagc attaatccag ccatgatggc
     2461 tgccgcccag gcagcactac agagcagttg gggtatgatg ggcatgttag ccagccagca
     2521 gaaccagtca ggcccatcgg gtaataacca aaaccaaggc aacatgcaga gggagccaaa
     2581 ccaggccttc ggttctggaa ataactctta tagtggctct aattctggtg cagcaattgg
     2641 tgggggatca gcatccaatg cagggtcggg cagtggtttt aatggaggct ttggctcaag
     2701 catggattct aagtcttctg gctggggaat gtagacccag ctttcttgta caaagtggtt
     2761 gatcctctac tagtaagctt gcatgcgata tctctagagc tgagaacttc agggtgagtt
     2821 tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg
     2881 gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct
     2941 catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt
     3001 tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta
     3061 aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag
     3121 gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta
     3181 aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg
     3241 gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc tttatggtta caatgatata
     3301 cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg
     3361 ttcatgcctt cttctctttc ctacagctcc tgggctgcaa tagtgtgttg gaattttttg
     3421 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3481 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     3541 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     3601 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     3661 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     3721 ccagtcatag ctgtccctct tctcttatgg agatccctcg actgcattaa tgaatcggcc
     3781 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5041 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5341 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     5401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5761 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     5821 actcgacgcg gccagcttgg tcgagctgga tacttcccgt ccgccagggg gacatgccgg
     5881 cgatgctgaa ggtcgcgcgc attcccgatg aagaggccgg ttaccgcctg ttgacctggt
     5941 gggacgggca gggcgccgag ttcataagag aagagggaca gctatgactg ggagtagtca
     6001 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     6061 aaaaaataac acaaaacaaa atataaaaaa aatctaacct caagtcaagg cttttctatg
     6121 gaataaggaa tggacagcag ggggctgttt catatactga tgacctcttt atagccaacc
     6181 tttgttcatg gcagccagca tatgggcata tgttgccaaa ctctaaacca aatactcatt
     6241 ctgatgtttt aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt
     6301 ccaacacact attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag
     6361 gggcttcatg atgtccccat aatttttggc agagggaaaa agatctcagt ggtatttgtg
     6421 agccagggca ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt
     6481 gaattctttg ccaaaatgat gagacagcac aacaaccagc acgttgccca ggagctgtag
     6541 gaaagagaag aaggcatgaa catggttagc agaggggccc ggtttggact cagagtattt
     6601 tatcctcatc tcaaacagtg tatatcattg taaccataaa gagaaaggca ggatgatgac
     6661 cagggtgtag ttgtttctac caataagaat atttccacgc cagccagaat ttatatgcag
     6721 aaatattcta ccttatcatt taattataac aattgttctc taaaactgtg ctgaagtaca
     6781 atataatata ccctgattgc cttgaaaaaa aagtgattag agaaagtact tacaatctga
     6841 caaataaaca aaagtgaatt taaaaattcg ttacaaatgc aagctaaagt ttaacgaaaa
     6901 agttacagaa aatgaaaaga aaataagagg agacaatggt tgtcaacaga gtagaaagtg
     6961 aaagaaacaa aattatcatg agggtccatg gtgatacaag ggacatcttc ccattctaaa
     7021 caacaccctg aaaactttgc cccctccata taacatgaat tttacaatag cgaaaaagaa
     7081 agaacaatca agggtcccca aactcaccct gaagttctca gtctagagtc gagaggatca
     7141 atcactttgt acaagaaagc tgggtctaga tagcttactt gtacagctcg tccatgccgc
     7201 cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt
     7261 gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct
     7321 tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc
     7381 tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct
     7441 cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca
     7501 gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca
     7561 cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct
     7621 tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact
     7681 gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg
     7741 cggtctgggt gccctcgtag gggcggccct gaggaaaaaa aaggaaaggg catgctcgcc
     7801 atgtgcaaga tacagtgttg aatgtggtgc aaaatataaa gatgtggaag aaaaagccac
     7861 tgaaaataat atgaggaaat aaaaaactag agtaaattat cagtggaaca atttctatta
     7921 tacaagacac tacacccata cattctccta atgctcatgt aagaatttct ccaaaaatac
     7981 cttccagcac tacaaactag aaaaaaaaag agacatggac accaaattaa gttcttaata
     8041 gtgaagaaca aaagaactac cttgcctgct ccagtggatc cagcaaccgc caacaactgt
     8101 cctctttcta tcttgaaatt aatatctttc aggacaggag taccaagaag tgagaaatta
     8161 ctgaagaaga ggctgtcatc accattagaa gtttttctat tgttattgtt ttgttttgct
     8221 ttctcaaata attccccaaa tccctgttaa aaaaaaacac acacacacac acacacacac
     8281 acatcaaaaa taaaagatga gtttgtcaga tattttcaat agatgcttgt tttatgtacc
     8341 atactcacct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     8401 tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctg
     8461 gttccggtgc ctccactttc atcttcacct ttagtgatac cggctgccgt cacgaactcc
     8521 ttcaggacca tgtgatcgcg cttctcgttc gggtctttgc tcagcgtggt ctgggtgctc
     8581 agatagtggt tgtccggcag gcgcaccggg ccgtcgccga tcggggtgtt ctgctgtttg
     8641 tggtcggctt cctgctggct gccgtcctca acgttgtggc ggatcttgaa gttagccttg
     8701 atgccgttct tctgcttgtc ggcttcgata cggacgttgt ggctgttgtg gttgtactcc
     8761 agcttgtgac ccaggatgtt gccgtcctcc ttgaagtcgt tgcccttcag ctcgatgcgg
     8821 ttcaccaggg tgtcgccctc gaacttcacc tccgcgcggg tcttgtaggt gccgtcgtcc
     8881 ttgaactcga tggtgcgctc ctggacgtag ccttccggca tggcggactt gaagaagtcg
     8941 tgacgcttca tgtggtccgg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     9001 acgagggtcg gccacggcac cggcagcttg ccggtggtgc agatgaactt cagggtcagc
     9061 ttgccctcgg tggcatcgcc ctcgccctcg ccacgcacgc tgaacttgtg gccgtttacg
     9121 tcgccgtcca gctcgacttt gataggcaca gtacctgtaa atagttcctc tcctttactg
     9181 gtaccatcat cagggaatga gcttacattt ctcctcttca gcatctggtc atccttctta
     9241 gctttcctca gctccacatt gacctctatt ctgcgacgcc tcatttctgt actgtctttt
     9301 cccttgttct tgaatctgtg aagacgggca gctggtgtat tagcattctc gttggtggat
     9361 cccatggtgg caccggtgaa ttctccaggc gatctgacgg ttcactaaac gagctctgct
     9421 tatataggcc tcccaccgta cacgccacct cgacatac
//
