LOCUS       sequence-231200-        9667 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 9667)
  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 9667)
  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: pHBS1401 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 F-S]
FEATURES             Location/Qualifiers
     source          1..9667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        346..549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     500..520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             617..1318
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1319..1384
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1409..1465
                     /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(2711..2735)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2790..3362
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3559..3614
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4012..4029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4183..4771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4263..4282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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     5565..5584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5803..5907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5975..5993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6468..6523
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6720..7292
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7638..7657)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7907..7925
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9547..9556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
       61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      541 agcagagctg gtttagtgaa ccgtcagatc cgctagggat cctctagtca gctgacgcgt
      601 taccaccggt gccaccatgg gaagcgagct gattaaggag aacatgcaca tgaagctgta
      661 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
      721 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
      781 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca
      841 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac
      901 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
      961 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca
     1021 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct
     1081 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc
     1141 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta
     1201 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca
     1261 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaattc
     1321 cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
     1381 atccaccgga tctagaggag gaagcggagc tactaacttc agcctgctga agcaggctgg
     1441 agacgtggag gagaaccctg gacctatgtc tgaatatatt cgggtaaccg aagatgagaa
     1501 cgatgagccc attgaaatac catcggaaga cgatgggacg gtgctgctct ccacggttac
     1561 agcccagttt ccaggggcgt gtgggcttcg ctacaggaat ccagtgtctc agtgtatgag
     1621 aggtgtccgg ctggtagaag gaattctgca tgccccagat gctggctggg gaaatctggt
     1681 gtatgttgtc aactatccaa aagataacaa aagaaaaatg gatgagacag atgcttcatc
     1741 agcagtgaaa gtgaaaagag cagtccagaa aacatccgat ttaatagtgt tgggtctccc
     1801 atggaaaaca accgaacagg acctgaaaga gtattttagt acctttggag aagttcttat
     1861 ggtgcaggtc aagaaagatc ttaagactgg tcattcaaag gggtttggct ttgttcgttt
     1921 tacggaatat gaaacacaag tgaaagtaat gtcacagcga catatgatag atggacgatg
     1981 gtgtgactgc aaacttccta attctaagca aagccaagat gagcctttga gaagcagaaa
     2041 agtgtttgtg gggcgctgta cagaggacat gactgaggat gagctgcggg agttcttctc
     2101 tcagtacggg gatgtgatgg atgtcttcat ccccaagcca ttcagggcct ttgcctttgt
     2161 tacatttgca gatgatcaga ttgcgcagtc tctttgtgga gaggacttga tcattaaagg
     2221 aatcagcgtt catatatcca atgccgaacc taaacataat tcaaaccggc agttggaaag
     2281 atcggggcgc agtggtggca acccaggcgg ttctggtaat cagggcggct cgggaaattc
     2341 cagaggcggg ggtgcagggc ttggcaacaa tcaggggtcc aatatgggcg gtggtatgaa
     2401 ctcaggtgcc agctcgatta atccggccat gatggcggca gcgcaggcgg cgctgcagtc
     2461 ttcctggggt atgatgggta tgttagcatc ccaacagaac cagagtggcc cgtcggggaa
     2521 taaccaaaat caggggaata tgcagcgcga accgaatcaa gcttcaggta gtggtaacaa
     2581 cagttatagt ggatcgaaca gtggcgccgc aattggatgg ggttccgcat ccaatgcagg
     2641 ctcaggcagc ggctcgaatg gcggctcagg ttctagtatg gattcgaaga gctctggttg
     2701 gggaatgtag acccagcttt cttgtacaaa gtggttgatc ctctactagt aagcttgcat
     2761 gcgatatctc tagagctgag aacttcaggg tgagtttggg gacccttgat tgttctttct
     2821 ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta
     2881 gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact
     2941 ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt
     3001 cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag
     3061 attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata ttatattgta
     3121 cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc
     3181 atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac accctggtca
     3241 tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat
     3301 actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc tctttcctac
     3361 agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc
     3421 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     3481 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     3541 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     3601 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     3661 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     3721 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     3781 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     3841 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     3901 tactcccagt catagctgtc cctcttctct tatggagatc cctcgactgc attaatgaat
     3961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4621 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5221 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     5581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5941 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6001 tcttcactcg acgcggccag cttggtcgag ctggatactt cccgtccgcc agggggacat
     6061 gccggcgatg ctgaaggtcg cgcgcattcc cgatgaagag gccggttacc gcctgttgac
     6121 ctggtgggac gggcagggcg ccgagttcat aagagaagag ggacagctat gactgggagt
     6181 agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt tagggatgtt
     6241 aaagaaaaaa ataacacaaa acaaaatata aaaaaaatct aacctcaagt caaggctttt
     6301 ctatggaata aggaatggac agcagggggc tgtttcatat actgatgacc tctttatagc
     6361 caacctttgt tcatggcagc cagcatatgg gcatatgttg ccaaactcta aaccaaatac
     6421 tcattctgat gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa
     6481 aaattccaac acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc
     6541 tcaaggggct tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat
     6601 ttgtgagcca gggcattggc cacaccagcc accaccttct gataggcagc ctgcacctga
     6661 ggagtgaatt ctttgccaaa atgatgagac agcacaacaa ccagcacgtt gcccaggagc
     6721 tgtaggaaag agaagaaggc atgaacatgg ttagcagagg ggcccggttt ggactcagag
     6781 tattttatcc tcatctcaaa cagtgtatat cattgtaacc ataaagagaa aggcaggatg
     6841 atgaccaggg tgtagttgtt tctaccaata agaatatttc cacgccagcc agaatttata
     6901 tgcagaaata ttctacctta tcatttaatt ataacaattg ttctctaaaa ctgtgctgaa
     6961 gtacaatata atataccctg attgccttga aaaaaaagtg attagagaaa gtacttacaa
     7021 tctgacaaat aaacaaaagt gaatttaaaa attcgttaca aatgcaagct aaagtttaac
     7081 gaaaaagtta cagaaaatga aaagaaaata agaggagaca atggttgtca acagagtaga
     7141 aagtgaaaga aacaaaatta tcatgagggt ccatggtgat acaagggaca tcttcccatt
     7201 ctaaacaaca ccctgaaaac tttgccccct ccatataaca tgaattttac aatagcgaaa
     7261 aagaaagaac aatcaagggt ccccaaactc accctgaagt tctcagtcta gagtcgagag
     7321 gatcaatcac tttgtacaag aaagctgggt ctagatagct tacttgtaca gctcgtccat
     7381 gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc
     7441 gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg
     7501 cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt
     7561 cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga
     7621 ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc
     7681 gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt
     7741 caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga
     7801 cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat
     7861 gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag
     7921 cttggcggtc tgggtgccct cgtaggggcg gccctgagga aaaaaaagga aagggcatgc
     7981 tcgccatgtg caagatacag tgttgaatgt ggtgcaaaat ataaagatgt ggaagaaaaa
     8041 gccactgaaa ataatatgag gaaataaaaa actagagtaa attatcagtg gaacaatttc
     8101 tattatacaa gacactacac ccatacattc tcctaatgct catgtaagaa tttctccaaa
     8161 aataccttcc agcactacaa actagaaaaa aaaagagaca tggacaccaa attaagttct
     8221 taatagtgaa gaacaaaaga actaccttgc ctgctccagt ggatccagca accgccaaca
     8281 actgtcctct ttctatcttg aaattaatat ctttcaggac aggagtacca agaagtgaga
     8341 aattactgaa gaagaggctg tcatcaccat tagaagtttt tctattgtta ttgttttgtt
     8401 ttgctttctc aaataattcc ccaaatccct gttaaaaaaa aacacacaca cacacacaca
     8461 cacacacatc aaaaataaaa gatgagtttg tcagatattt tcaatagatg cttgttttat
     8521 gtaccatact cacctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     8581 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
     8641 tgctggttcc ggtgcctcca ctttcatctt cacctttagt gataccggct gccgtcacga
     8701 actccttcag gaccatgtga tcgcgcttct cgttcgggtc tttgctcagc gtggtctggg
     8761 tgctcagata gtggttgtcc ggcaggcgca ccgggccgtc gccgatcggg gtgttctgct
     8821 gtttgtggtc ggcttcctgc tggctgccgt cctcaacgtt gtggcggatc ttgaagttag
     8881 ccttgatgcc gttcttctgc ttgtcggctt cgatacggac gttgtggctg ttgtggttgt
     8941 actccagctt gtgacccagg atgttgccgt cctccttgaa gtcgttgccc ttcagctcga
     9001 tgcggttcac cagggtgtcg ccctcgaact tcacctccgc gcgggtcttg taggtgccgt
     9061 cgtccttgaa ctcgatggtg cgctcctgga cgtagccttc cggcatggcg gacttgaaga
     9121 agtcgtgacg cttcatgtgg tccgggtagc ggctgaagca ctgcacgccg taggtcaggg
     9181 tggtcacgag ggtcggccac ggcaccggca gcttgccggt ggtgcagatg aacttcaggg
     9241 tcagcttgcc ctcggtggca tcgccctcgc cctcgccacg cacgctgaac ttgtggccgt
     9301 ttacgtcgcc gtccagctcg actttgatag gcacagtacc tgtaaatagt tcctctcctt
     9361 tactggtacc atcatcaggg aatgagctta catttctcct cttcagcatc tggtcatcct
     9421 tcttagcttt cctcagctcc acattgacct ctattctgcg acgcctcatt tctgtactgt
     9481 cttttccctt gttcttgaat ctgtgaagac gggcagctgg tgtattagca ttctcgttgg
     9541 tggatcccat ggtggcaccg gtgaattctc caggcgatct gacggttcac taaacgagct
     9601 ctgcttatat aggcctccca ccgtacacgc cacctcgaca tactcgagta gttattaata
     9661 gtaatca
//
