LOCUS       sequence-261806-        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: pHBS1506 [IBB-GFP-mCherry3E]-[BFP-TDP43 FYW-L]
FEATURES             Location/Qualifiers
     source          1..9498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    173..228
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(703..721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        789..893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(894..962,963..1754)
                     /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(1112..1131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1925..2513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2414..2433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2667..2684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3082..3137
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          3334..3906
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4252..4271)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4521..4539
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6161..6170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        6323..6626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6627..6830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6781..6801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6892..6901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6898..7599
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(8851..8875)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          8902..9474
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 gtgctgtctc atcattttgg caaagaattc actcctcagg tgcaggctgc ctatcagaag
       61 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
      121 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
      181 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
      241 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
      301 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
      361 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
      421 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
      481 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
      541 ctcttatgaa ctcggcgccc tgcccgtccc accaggtcaa caggcggtaa ccggcctctt
      601 catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc cccctggcgg acgggaagta
      661 tccagctcga ccaagctggc cgcgtcgagt gaagacgaaa gggcctcgtg atacgcctat
      721 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2041 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2461 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2581 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2641 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2701 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagt cgagggatct
     2761 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     2821 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     2881 tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg
     2941 gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat
     3001 atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     3061 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     3121 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     3181 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     3241 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc caaaatgatg
     3301 agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga aggcatgaac
     3361 atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct caaacagtgt
     3421 atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt tgtttctacc
     3481 aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac cttatcattt
     3541 aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac cctgattgcc
     3601 ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa aagtgaattt
     3661 aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa atgaaaagaa
     3721 aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa attatcatga
     3781 gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga aaactttgcc
     3841 ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa gggtccccaa
     3901 actcaccctg aagttctcag tctagagtcg agaggatcaa tcactttgta caagaaagct
     3961 gggtctagat agcttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg
     4021 cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga
     4081 tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga
     4141 cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca
     4201 gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct
     4261 gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact
     4321 cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca
     4381 tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg
     4441 cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg
     4501 cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg
     4561 ggcggccctg aggaaaaaaa aggaaagggc atgctcgcca tgtgcaagat acagtgttga
     4621 atgtggtgca aaatataaag atgtggaaga aaaagccact gaaaataata tgaggaaata
     4681 aaaaactaga gtaaattatc agtggaacaa tttctattat acaagacact acacccatac
     4741 attctcctaa tgctcatgta agaatttctc caaaaatacc ttccagcact acaaactaga
     4801 aaaaaaaaga gacatggaca ccaaattaag ttcttaatag tgaagaacaa aagaactacc
     4861 ttgcctgctc cagtggatcc agcaaccgcc aacaactgtc ctctttctat cttgaaatta
     4921 atatctttca ggacaggagt accaagaagt gagaaattac tgaagaagag gctgtcatca
     4981 ccattagaag tttttctatt gttattgttt tgttttgctt tctcaaataa ttccccaaat
     5041 ccctgttaaa aaaaaacaca cacacacaca cacacacaca catcaaaaat aaaagatgag
     5101 tttgtcagat attttcaata gatgcttgtt ttatgtacca tactcacctc gccctcgccc
     5161 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     5221 tccttgatga tggccatgtt atcctcctcg cccttgctgg ttccggtgcc tccactttca
     5281 tcttcacctt tagtgatacc ggctgccgtc acgaactcct tcaggaccat gtgatcgcgc
     5341 ttctcgttcg ggtctttgct cagcgtggtc tgggtgctca gatagtggtt gtccggcagg
     5401 cgcaccgggc cgtcgccgat cggggtgttc tgctgtttgt ggtcggcttc ctgctggctg
     5461 ccgtcctcaa cgttgtggcg gatcttgaag ttagccttga tgccgttctt ctgcttgtcg
     5521 gcttcgatac ggacgttgtg gctgttgtgg ttgtactcca gcttgtgacc caggatgttg
     5581 ccgtcctcct tgaagtcgtt gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     5641 aacttcacct ccgcgcgggt cttgtaggtg ccgtcgtcct tgaactcgat ggtgcgctcc
     5701 tggacgtagc cttccggcat ggcggacttg aagaagtcgt gacgcttcat gtggtccggg
     5761 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtcgg ccacggcacc
     5821 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccctcggt ggcatcgccc
     5881 tcgccctcgc cacgcacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgactttg
     5941 ataggcacag tacctgtaaa tagttcctct cctttactgg taccatcatc agggaatgag
     6001 cttacatttc tcctcttcag catctggtca tccttcttag ctttcctcag ctccacattg
     6061 acctctattc tgcgacgcct catttctgta ctgtcttttc ccttgttctt gaatctgtga
     6121 agacgggcag ctggtgtatt agcattctcg ttggtggatc ccatggtggc accggtgaat
     6181 tctccaggcg atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac
     6241 acgccacctc gacatactcg agtagttatt aatagtaatc aattacgggg tcattagttc
     6301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     6661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     6841 accgtcagat ccgctaggga tcctctagtc agctgacgcg ttaccaccgg tgccaccatg
     6901 ggaagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     6961 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     7021 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     7081 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     7141 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     7201 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     7261 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     7321 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     7381 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     7441 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     7501 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     7561 tactgcgacc tccctagcaa actggggcac aagcttaatt ccggaatgtc tgaatatatt
     7621 cgggtaaccg aagatgagaa cgatgagccc attgaaatac catcggaaga cgatgggacg
     7681 gtgctgctct ccacggttac agcccagttt ccaggggcgt gtgggcttcg ctacaggaat
     7741 ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag gaattctgca tgccccagat
     7801 gctggctggg gaaatctggt gtatgttgtc aactatccaa aagataacaa aagaaaaatg
     7861 gatgagacag atgcttcatc agcagtgaaa gtgaaaagag cagtccagaa aacatccgat
     7921 ttaatagtgt tgggtctccc atggaaaaca accgaacagg acctgaaaga gtattttagt
     7981 acctttggag aagttcttat ggtgcaggtc aagaaagatc ttaagactgg tcattcaaag
     8041 gggtttggct ttgttcgttt tacggaatat gaaacacaag tgaaagtaat gtcacagcga
     8101 catatgatag atggacgatg gtgtgactgc aaacttccta attctaagca aagccaagat
     8161 gagcctttga gaagcagaaa agtgtttgtg gggcgctgta cagaggacat gactgaggat
     8221 gagctgcggg agttcttctc tcagtacggg gatgtgatgg atgtcttcat ccccaagcca
     8281 ttcagggcct ttgcctttgt tacatttgca gatgatcaga ttgcgcagtc tctttgtgga
     8341 gaggacttga tcattaaagg aatcagcgtt catatatcca atgccgaacc taagcacaac
     8401 tcaaaccggc agctggaacg ctcaggccgc ctgggaggta atcctggagg actcggtaac
     8461 caaggtggcc tcggaaacag caggggaggc ggtgcaggac tgggcaacaa tcaagggagt
     8521 aacatgggtg ggggcatgaa ccttggcgct cttagcatca accccgcgat gatggcggca
     8581 gctcaggctg cccttcaaag tagttggggt atgatgggta tgctggcgag ccagcagaac
     8641 cagagcggcc cctccgggaa caaccaaaac cagggtaaca tgcagcgcga accgaatcag
     8701 gctctggggt ccggtaacaa ctccctgtcc ggtagtaatt caggagctgc tattggtctg
     8761 gggtcagcca gtaatgcagg aagcgggtct ggcctgaatg gggggctggg aagtagcatg
     8821 gatagtaagt cttcaggact tggaatgtag acccagcttt cttgtacaaa gtggttgatc
     8881 ctctagactg agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct
     8941 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
     9001 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
     9061 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
     9121 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
     9181 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
     9241 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
     9301 tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg
     9361 cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag
     9421 tccaaaccgg gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg
     9481 ggcaacgtgc tggttgtt
//
