LOCUS       sequence-261749-        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: pHBS1504 [IBB-GFP-mCherry3E]-[BFP-TDP43 F-S]
FEATURES             Location/Qualifiers
     source          1..9498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    279..334
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(809..827)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        895..999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1000..1068,1069..1860)
                     /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(1218..1237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2031..2619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2520..2539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2773..2790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3188..3243
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          3440..4012
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4358..4377)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4627..4645
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6267..6276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        6429..6732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6733..6936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6887..6907
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6998..7007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7004..7705
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(8957..8981)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          join(9008..9498,1..82)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
       61 tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
      121 ttttggcaaa gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
      181 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
      241 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
      301 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
      361 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
      421 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
      481 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
      541 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
      601 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaactcg
      661 gcgccctgcc cgtcccacca ggtcaacagg cggtaaccgg cctcttcatc gggaatgcgc
      721 gcgaccttca gcatcgccgg catgtccccc tggcggacgg gaagtatcca gctcgaccaa
      781 gctggccgcg tcgagtgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      841 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      961 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1021 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1081 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1141 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1201 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1261 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1321 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1381 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1441 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1501 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1561 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1621 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1681 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1741 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1801 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1861 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1921 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1981 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2041 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2101 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2161 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2221 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2281 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2341 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2401 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2461 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2521 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2581 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2641 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2701 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2761 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2821 cctctccccg cgcgttggcc gattcattaa tgcagtcgag ggatctccat aagagaagag
     2881 ggacagctat gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta
     2941 aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa acaaaatata aaaaaaatct
     3001 aacctcaagt caaggctttt ctatggaata aggaatggac agcagggggc tgtttcatat
     3061 actgatgacc tctttatagc caacctttgt tcatggcagc cagcatatgg gcatatgttg
     3121 ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc
     3181 ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt
     3241 attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg
     3301 gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct
     3361 gataggcagc ctgcacctga ggagtgaatt ctttgccaaa atgatgagac agcacaacaa
     3421 ccagcacgtt gcccaggagc tgtaggaaag agaagaaggc atgaacatgg ttagcagagg
     3481 ggcccggttt ggactcagag tattttatcc tcatctcaaa cagtgtatat cattgtaacc
     3541 ataaagagaa aggcaggatg atgaccaggg tgtagttgtt tctaccaata agaatatttc
     3601 cacgccagcc agaatttata tgcagaaata ttctacctta tcatttaatt ataacaattg
     3661 ttctctaaaa ctgtgctgaa gtacaatata atataccctg attgccttga aaaaaaagtg
     3721 attagagaaa gtacttacaa tctgacaaat aaacaaaagt gaatttaaaa attcgttaca
     3781 aatgcaagct aaagtttaac gaaaaagtta cagaaaatga aaagaaaata agaggagaca
     3841 atggttgtca acagagtaga aagtgaaaga aacaaaatta tcatgagggt ccatggtgat
     3901 acaagggaca tcttcccatt ctaaacaaca ccctgaaaac tttgccccct ccatataaca
     3961 tgaattttac aatagcgaaa aagaaagaac aatcaagggt ccccaaactc accctgaagt
     4021 tctcagtcta gagtcgagag gatcaatcac tttgtacaag aaagctgggt ctagatagct
     4081 tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt
     4141 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag
     4201 gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag
     4261 tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg
     4321 gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg
     4381 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg
     4441 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac
     4501 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg
     4561 taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg
     4621 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctgagga
     4681 aaaaaaagga aagggcatgc tcgccatgtg caagatacag tgttgaatgt ggtgcaaaat
     4741 ataaagatgt ggaagaaaaa gccactgaaa ataatatgag gaaataaaaa actagagtaa
     4801 attatcagtg gaacaatttc tattatacaa gacactacac ccatacattc tcctaatgct
     4861 catgtaagaa tttctccaaa aataccttcc agcactacaa actagaaaaa aaaagagaca
     4921 tggacaccaa attaagttct taatagtgaa gaacaaaaga actaccttgc ctgctccagt
     4981 ggatccagca accgccaaca actgtcctct ttctatcttg aaattaatat ctttcaggac
     5041 aggagtacca agaagtgaga aattactgaa gaagaggctg tcatcaccat tagaagtttt
     5101 tctattgtta ttgttttgtt ttgctttctc aaataattcc ccaaatccct gttaaaaaaa
     5161 aacacacaca cacacacaca cacacacatc aaaaataaaa gatgagtttg tcagatattt
     5221 tcaatagatg cttgttttat gtaccatact cacctcgccc tcgccctcga tctcgaactc
     5281 gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc
     5341 catgttatcc tcctcgccct tgctggttcc ggtgcctcca ctttcatctt cacctttagt
     5401 gataccggct gccgtcacga actccttcag gaccatgtga tcgcgcttct cgttcgggtc
     5461 tttgctcagc gtggtctggg tgctcagata gtggttgtcc ggcaggcgca ccgggccgtc
     5521 gccgatcggg gtgttctgct gtttgtggtc ggcttcctgc tggctgccgt cctcaacgtt
     5581 gtggcggatc ttgaagttag ccttgatgcc gttcttctgc ttgtcggctt cgatacggac
     5641 gttgtggctg ttgtggttgt actccagctt gtgacccagg atgttgccgt cctccttgaa
     5701 gtcgttgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctccgc
     5761 gcgggtcttg taggtgccgt cgtccttgaa ctcgatggtg cgctcctgga cgtagccttc
     5821 cggcatggcg gacttgaaga agtcgtgacg cttcatgtgg tccgggtagc ggctgaagca
     5881 ctgcacgccg taggtcaggg tggtcacgag ggtcggccac ggcaccggca gcttgccggt
     5941 ggtgcagatg aacttcaggg tcagcttgcc ctcggtggca tcgccctcgc cctcgccacg
     6001 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg actttgatag gcacagtacc
     6061 tgtaaatagt tcctctcctt tactggtacc atcatcaggg aatgagctta catttctcct
     6121 cttcagcatc tggtcatcct tcttagcttt cctcagctcc acattgacct ctattctgcg
     6181 acgcctcatt tctgtactgt cttttccctt gttcttgaat ctgtgaagac gggcagctgg
     6241 tgtattagca ttctcgttgg tggatcccat ggtggcaccg gtgaattctc caggcgatct
     6301 gacggttcac taaacgagct ctgcttatat aggcctccca ccgtacacgc cacctcgaca
     6361 tactcgagta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6421 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6481 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6601 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6661 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6721 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6781 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6841 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6901 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     6961 tagggatcct ctagtcagct gacgcgttac caccggtgcc accatgggaa gcgagctgat
     7021 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
     7081 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     7141 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
     7201 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
     7261 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     7321 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     7381 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
     7441 gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
     7501 gggcgggagc catctgatcg caaacgccaa gaccacatat agatccaaga aacccgctaa
     7561 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     7621 caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
     7681 tagcaaactg gggcacaagc ttaattccgg aatgtctgaa tatattcggg taaccgaaga
     7741 tgagaacgat gagcccattg aaataccatc ggaagacgat gggacggtgc tgctctccac
     7801 ggttacagcc cagtttccag gggcgtgtgg gcttcgctac aggaatccag tgtctcagtg
     7861 tatgagaggt gtccggctgg tagaaggaat tctgcatgcc ccagatgctg gctggggaaa
     7921 tctggtgtat gttgtcaact atccaaaaga taacaaaaga aaaatggatg agacagatgc
     7981 ttcatcagca gtgaaagtga aaagagcagt ccagaaaaca tccgatttaa tagtgttggg
     8041 tctcccatgg aaaacaaccg aacaggacct gaaagagtat tttagtacct ttggagaagt
     8101 tcttatggtg caggtcaaga aagatcttaa gactggtcat tcaaaggggt ttggctttgt
     8161 tcgttttacg gaatatgaaa cacaagtgaa agtaatgtca cagcgacata tgatagatgg
     8221 acgatggtgt gactgcaaac ttcctaattc taagcaaagc caagatgagc ctttgagaag
     8281 cagaaaagtg tttgtggggc gctgtacaga ggacatgact gaggatgagc tgcgggagtt
     8341 cttctctcag tacggggatg tgatggatgt cttcatcccc aagccattca gggcctttgc
     8401 ctttgttaca tttgcagatg atcagattgc gcagtctctt tgtggagagg acttgatcat
     8461 taaaggaatc agcgttcata tatccaatgc cgaacctaaa cataattcaa accggcagtt
     8521 ggaaagatcg gggcgcagtg gtggcaaccc aggcggttct ggtaatcagg gcggctcggg
     8581 aaattccaga ggcgggggtg cagggcttgg caacaatcag gggtccaata tgggcggtgg
     8641 tatgaactca ggtgccagct cgattaatcc ggccatgatg gcggcagcgc aggcggcgct
     8701 gcagtcttcc tggggtatga tgggtatgtt agcatcccaa cagaaccaga gtggcccgtc
     8761 ggggaataac caaaatcagg ggaatatgca gcgcgaaccg aatcaagctt caggtagtgg
     8821 taacaacagt tatagtggat cgaacagtgg cgccgcaatt ggatggggtt ccgcatccaa
     8881 tgcaggctca ggcagcggct cgaatggcgg ctcaggttct agtatggatt cgaagagctc
     8941 tggttgggga atgtagaccc agctttcttg tacaaagtgg ttgatcctct agactgagaa
     9001 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     9061 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     9121 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     9181 gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt
     9241 gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc
     9301 actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac
     9361 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     9421 aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt tctctttatg
     9481 gttacaatga tatacact
//
