LOCUS       sequence-261743-        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: pHBS1508 [IBB-GFP-mCherry 3E]-[BFP-TDP43 VLIM-F]
FEATURES             Location/Qualifiers
     source          1..9498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    293..348
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(823..841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        909..1013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1014..1082,1083..1874)
                     /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(1232..1251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2045..2633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2534..2553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2787..2804
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3202..3257
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          3454..4026
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4372..4391)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4641..4659
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6281..6290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        6443..6746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6747..6950
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6901..6921
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7012..7021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7018..7719
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(8971..8995)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          join(9022..9498,1..96)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
       61 gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt gctggttgtt
      121 gtgctgtctc atcattttgg caaagaattc actcctcagg tgcaggctgc ctatcagaag
      181 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
      241 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
      301 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
      361 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
      421 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
      481 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
      541 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
      601 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
      661 ctcttatgaa ctcggcgccc tgcccgtccc accaggtcaa caggcggtaa ccggcctctt
      721 catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc cccctggcgg acgggaagta
      781 tccagctcga ccaagctggc cgcgtcgagt gaagacgaaa gggcctcgtg atacgcctat
      841 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1201 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1321 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1561 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1621 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1681 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1741 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1801 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1861 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1921 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1981 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2041 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2101 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2161 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2221 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2281 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2341 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2401 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2461 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2521 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2581 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2641 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2701 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2761 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2821 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagt cgagggatct
     2881 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     2941 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     3001 tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg
     3061 gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat
     3121 atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     3181 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     3241 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     3301 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     3361 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc caaaatgatg
     3421 agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga aggcatgaac
     3481 atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct caaacagtgt
     3541 atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt tgtttctacc
     3601 aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac cttatcattt
     3661 aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac cctgattgcc
     3721 ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa aagtgaattt
     3781 aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa atgaaaagaa
     3841 aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa attatcatga
     3901 gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga aaactttgcc
     3961 ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa gggtccccaa
     4021 actcaccctg aagttctcag tctagagtcg agaggatcaa tcactttgta caagaaagct
     4081 gggtctagat agcttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg
     4141 cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga
     4201 tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga
     4261 cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca
     4321 gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct
     4381 gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact
     4441 cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca
     4501 tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg
     4561 cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg
     4621 cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg
     4681 ggcggccctg aggaaaaaaa aggaaagggc atgctcgcca tgtgcaagat acagtgttga
     4741 atgtggtgca aaatataaag atgtggaaga aaaagccact gaaaataata tgaggaaata
     4801 aaaaactaga gtaaattatc agtggaacaa tttctattat acaagacact acacccatac
     4861 attctcctaa tgctcatgta agaatttctc caaaaatacc ttccagcact acaaactaga
     4921 aaaaaaaaga gacatggaca ccaaattaag ttcttaatag tgaagaacaa aagaactacc
     4981 ttgcctgctc cagtggatcc agcaaccgcc aacaactgtc ctctttctat cttgaaatta
     5041 atatctttca ggacaggagt accaagaagt gagaaattac tgaagaagag gctgtcatca
     5101 ccattagaag tttttctatt gttattgttt tgttttgctt tctcaaataa ttccccaaat
     5161 ccctgttaaa aaaaaacaca cacacacaca cacacacaca catcaaaaat aaaagatgag
     5221 tttgtcagat attttcaata gatgcttgtt ttatgtacca tactcacctc gccctcgccc
     5281 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     5341 tccttgatga tggccatgtt atcctcctcg cccttgctgg ttccggtgcc tccactttca
     5401 tcttcacctt tagtgatacc ggctgccgtc acgaactcct tcaggaccat gtgatcgcgc
     5461 ttctcgttcg ggtctttgct cagcgtggtc tgggtgctca gatagtggtt gtccggcagg
     5521 cgcaccgggc cgtcgccgat cggggtgttc tgctgtttgt ggtcggcttc ctgctggctg
     5581 ccgtcctcaa cgttgtggcg gatcttgaag ttagccttga tgccgttctt ctgcttgtcg
     5641 gcttcgatac ggacgttgtg gctgttgtgg ttgtactcca gcttgtgacc caggatgttg
     5701 ccgtcctcct tgaagtcgtt gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     5761 aacttcacct ccgcgcgggt cttgtaggtg ccgtcgtcct tgaactcgat ggtgcgctcc
     5821 tggacgtagc cttccggcat ggcggacttg aagaagtcgt gacgcttcat gtggtccggg
     5881 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtcgg ccacggcacc
     5941 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccctcggt ggcatcgccc
     6001 tcgccctcgc cacgcacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgactttg
     6061 ataggcacag tacctgtaaa tagttcctct cctttactgg taccatcatc agggaatgag
     6121 cttacatttc tcctcttcag catctggtca tccttcttag ctttcctcag ctccacattg
     6181 acctctattc tgcgacgcct catttctgta ctgtcttttc ccttgttctt gaatctgtga
     6241 agacgggcag ctggtgtatt agcattctcg ttggtggatc ccatggtggc accggtgaat
     6301 tctccaggcg atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac
     6361 acgccacctc gacatactcg agtagttatt aatagtaatc aattacgggg tcattagttc
     6421 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6481 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6541 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6601 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6661 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6721 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     6781 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6841 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6901 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     6961 accgtcagat ccgctaggga tcctctagtc agctgacgcg ttaccaccgg tgccaccatg
     7021 ggaagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     7081 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     7141 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     7201 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     7261 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     7321 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     7381 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     7441 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     7501 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     7561 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     7621 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     7681 tactgcgacc tccctagcaa actggggcac aagcttaatt ccggaatgtc tgaatatatt
     7741 cgggtaaccg aagatgagaa cgatgagccc attgaaatac catcggaaga cgatgggacg
     7801 gtgctgctct ccacggttac agcccagttt ccaggggcgt gtgggcttcg ctacaggaat
     7861 ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag gaattctgca tgccccagat
     7921 gctggctggg gaaatctggt gtatgttgtc aactatccaa aagataacaa aagaaaaatg
     7981 gatgagacag atgcttcatc agcagtgaaa gtgaaaagag cagtccagaa aacatccgat
     8041 ttaatagtgt tgggtctccc atggaaaaca accgaacagg acctgaaaga gtattttagt
     8101 acctttggag aagttcttat ggtgcaggtc aagaaagatc ttaagactgg tcattcaaag
     8161 gggtttggct ttgttcgttt tacggaatat gaaacacaag tgaaagtaat gtcacagcga
     8221 catatgatag atggacgatg gtgtgactgc aaacttccta attctaagca aagccaagat
     8281 gagcctttga gaagcagaaa agtgtttgtg gggcgctgta cagaggacat gactgaggat
     8341 gagctgcggg agttcttctc tcagtacggg gatgtgatgg atgtcttcat ccccaagcca
     8401 ttcagggcct ttgcctttgt tacatttgca gatgatcaga ttgcgcagtc tctttgtgga
     8461 gaggacttga tcattaaagg aatcagcgtt catatatcca atgccgaacc taagcataac
     8521 agtaatcggc aatttgagcg atccggcaga tttggaggga accctggagg attcggcaac
     8581 caggggggct ttggaaattc aagaggaggg ggtgcgggat tcggtaacaa ccaggggtcc
     8641 aatttcggtg gggggttcaa ctttggggcg ttttctttca atcccgctat gatggctgcg
     8701 gctcaagcgg ctctgcaaag tagctggggc atgatgggta tgctcgcgtc acaacagaat
     8761 cagagtggac caagtggaaa taatcagaat caagggaatt ttcaacgaga acctaaccaa
     8821 gcattcggat ctggtaataa tagctactcc gggagtaact ccggagcagc tttcgggtgg
     8881 ggctccgcgt caaacgctgg cagcggctct ggattcaacg ggggattcgg aagcagtttc
     8941 gattcaaaaa gcagtggctg ggggttctag acccagcttt cttgtacaaa gtggttgatc
     9001 ctctagactg agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct
     9061 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
     9121 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
     9181 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
     9241 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
     9301 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
     9361 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
     9421 tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg
     9481 cctttctctt tatggtta
//
