LOCUS       sequence-261759-        9498 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 9498)
  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 9498)
  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: pHBS1527 [IBB-GFP-mCherry3E]-[BFP-TDP43 W-S]
FEATURES             Location/Qualifiers
     source          1..9498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      570..579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             576..1277
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(2529..2553)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2580..3152
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3349..3404
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3879..3897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4070..4138,4139..4930)
                     /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     complement(4288..4307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5101..5689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5590..5609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5843..5860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6258..6313
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6510..7082
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7428..7447)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7697..7715
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9337..9346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagggatc
      541 ctctagtcag ctgacgcgtt accaccggtg ccaccatggg aagcgagctg attaaggaga
      601 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
      661 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
      721 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
      781 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
      841 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
      901 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
      961 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt
     1021 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
     1081 gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct aagaacctca
     1141 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
     1201 agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac
     1261 tggggcacaa gcttaattcc ggaatgtctg aatatattcg ggtaaccgaa gatgagaacg
     1321 atgagcccat tgaaatacca tcggaagacg atgggacggt gctgctctcc acggttacag
     1381 cccagtttcc aggggcgtgt gggcttcgct acaggaatcc agtgtctcag tgtatgagag
     1441 gtgtccggct ggtagaagga attctgcatg ccccagatgc tggctgggga aatctggtgt
     1501 atgttgtcaa ctatccaaaa gataacaaaa gaaaaatgga tgagacagat gcttcatcag
     1561 cagtgaaagt gaaaagagca gtccagaaaa catccgattt aatagtgttg ggtctcccat
     1621 ggaaaacaac cgaacaggac ctgaaagagt attttagtac ctttggagaa gttcttatgg
     1681 tgcaggtcaa gaaagatctt aagactggtc attcaaaggg gtttggcttt gttcgtttta
     1741 cggaatatga aacacaagtg aaagtaatgt cacagcgaca tatgatagat ggacgatggt
     1801 gtgactgcaa acttcctaat tctaagcaaa gccaagatga gcctttgaga agcagaaaag
     1861 tgtttgtggg gcgctgtaca gaggacatga ctgaggatga gctgcgggag ttcttctctc
     1921 agtacgggga tgtgatggat gtcttcatcc ccaagccatt cagggccttt gcctttgtta
     1981 catttgcaga tgatcagatt gcgcagtctc tttgtggaga ggacttgatc attaaaggaa
     2041 tcagcgttca tatatccaat gccgaaccta agcataacag caatcggcag ttagaaagat
     2101 cgggacgatt tgggggcaac ccaggaggtt ttgggaatca agggggcttt ggcaatagtc
     2161 gtggcggtgg tgcagggctg ggtaataatc aagggtctaa catgggaggt ggtatgaatt
     2221 tcggcgcgtt cagcattaat cccgcgatga tggccgccgc gcaagcagcc ttacagtcgt
     2281 cttgggggat gatgggaatg ctggcttctc agcagaatca gagcggtccg tcgggaaaca
     2341 atcaaaacca ggggaatatg cagcgtgaac cgaatcaggc attcgggagt ggcaacaatt
     2401 cttactccgg gtctaatagt ggtgcagcca ttggtagtgg ttccgcctcc aatgcgggtt
     2461 cgggaagcgg ctttaacggc ggctttggat catctatgga ttcaaaatca tcaggatcgg
     2521 gaatgtagac ccagctttct tgtacaaagt ggttgatcct ctagactgag aacttcaggg
     2581 tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg
     2641 gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg
     2701 gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc
     2761 ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa
     2821 tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt
     2881 ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     2941 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     3001 ttattggtag aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat
     3061 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     3121 accatgttca tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc
     3181 tgtctcatca ttttggcaaa gaattcactc ctcaggtgca ggctgcctat cagaaggtgg
     3241 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     3301 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     3361 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     3421 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     3481 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     3541 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     3601 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     3661 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     3721 tatgaactcg gcgccctgcc cgtcccacca ggtcaacagg cggtaaccgg cctcttcatc
     3781 gggaatgcgc gcgaccttca gcatcgccgg catgtccccc tggcggacgg gaagtatcca
     3841 gctcgaccaa gctggccgcg tcgagtgaag acgaaagggc ctcgtgatac gcctattttt
     3901 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     4321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5761 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5821 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5881 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtcgag ggatctccat
     5941 aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa tctggctagt
     6001 aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa acaaaatata
     6061 aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac agcagggggc
     6121 tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc cagcatatgg
     6181 gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc
     6241 ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata
     6301 aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt
     6361 ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc
     6421 accaccttct gataggcagc ctgcacctga ggagtgaatt ctttgccaaa atgatgagac
     6481 agcacaacaa ccagcacgtt gcccaggagc tgtaggaaag agaagaaggc atgaacatgg
     6541 ttagcagagg ggcccggttt ggactcagag tattttatcc tcatctcaaa cagtgtatat
     6601 cattgtaacc ataaagagaa aggcaggatg atgaccaggg tgtagttgtt tctaccaata
     6661 agaatatttc cacgccagcc agaatttata tgcagaaata ttctacctta tcatttaatt
     6721 ataacaattg ttctctaaaa ctgtgctgaa gtacaatata atataccctg attgccttga
     6781 aaaaaaagtg attagagaaa gtacttacaa tctgacaaat aaacaaaagt gaatttaaaa
     6841 attcgttaca aatgcaagct aaagtttaac gaaaaagtta cagaaaatga aaagaaaata
     6901 agaggagaca atggttgtca acagagtaga aagtgaaaga aacaaaatta tcatgagggt
     6961 ccatggtgat acaagggaca tcttcccatt ctaaacaaca ccctgaaaac tttgccccct
     7021 ccatataaca tgaattttac aatagcgaaa aagaaagaac aatcaagggt ccccaaactc
     7081 accctgaagt tctcagtcta gagtcgagag gatcaatcac tttgtacaag aaagctgggt
     7141 ctagatagct tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg
     7201 ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt
     7261 gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc
     7321 agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc
     7381 gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat
     7441 tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc
     7501 gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac
     7561 gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg
     7621 gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa
     7681 gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg
     7741 gccctgagga aaaaaaagga aagggcatgc tcgccatgtg caagatacag tgttgaatgt
     7801 ggtgcaaaat ataaagatgt ggaagaaaaa gccactgaaa ataatatgag gaaataaaaa
     7861 actagagtaa attatcagtg gaacaatttc tattatacaa gacactacac ccatacattc
     7921 tcctaatgct catgtaagaa tttctccaaa aataccttcc agcactacaa actagaaaaa
     7981 aaaagagaca tggacaccaa attaagttct taatagtgaa gaacaaaaga actaccttgc
     8041 ctgctccagt ggatccagca accgccaaca actgtcctct ttctatcttg aaattaatat
     8101 ctttcaggac aggagtacca agaagtgaga aattactgaa gaagaggctg tcatcaccat
     8161 tagaagtttt tctattgtta ttgttttgtt ttgctttctc aaataattcc ccaaatccct
     8221 gttaaaaaaa aacacacaca cacacacaca cacacacatc aaaaataaaa gatgagtttg
     8281 tcagatattt tcaatagatg cttgttttat gtaccatact cacctcgccc tcgccctcga
     8341 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     8401 tgatgatggc catgttatcc tcctcgccct tgctggttcc ggtgcctcca ctttcatctt
     8461 cacctttagt gataccggct gccgtcacga actccttcag gaccatgtga tcgcgcttct
     8521 cgttcgggtc tttgctcagc gtggtctggg tgctcagata gtggttgtcc ggcaggcgca
     8581 ccgggccgtc gccgatcggg gtgttctgct gtttgtggtc ggcttcctgc tggctgccgt
     8641 cctcaacgtt gtggcggatc ttgaagttag ccttgatgcc gttcttctgc ttgtcggctt
     8701 cgatacggac gttgtggctg ttgtggttgt actccagctt gtgacccagg atgttgccgt
     8761 cctccttgaa gtcgttgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     8821 tcacctccgc gcgggtcttg taggtgccgt cgtccttgaa ctcgatggtg cgctcctgga
     8881 cgtagccttc cggcatggcg gacttgaaga agtcgtgacg cttcatgtgg tccgggtagc
     8941 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtcggccac ggcaccggca
     9001 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc ctcggtggca tcgccctcgc
     9061 cctcgccacg cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg actttgatag
     9121 gcacagtacc tgtaaatagt tcctctcctt tactggtacc atcatcaggg aatgagctta
     9181 catttctcct cttcagcatc tggtcatcct tcttagcttt cctcagctcc acattgacct
     9241 ctattctgcg acgcctcatt tctgtactgt cttttccctt gttcttgaat ctgtgaagac
     9301 gggcagctgg tgtattagca ttctcgttgg tggatcccat ggtggcaccg gtgaattctc
     9361 caggcgatct gacggttcac taaacgagct ctgcttatat aggcctccca ccgtacacgc
     9421 cacctcgaca tactcgagta gttattaata gtaatcaatt acggggtcat tagttcatag
     9481 cccatatatg gagttccg
//
