LOCUS       sequence-231310-        9639 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 9639)
  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 9639)
  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: pHBS1405 [IBB-GFP-mCherry 3E]-[BFP-P2A-TDP43 V...
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             634..1335
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1336..1401
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1426..1482
                     /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(2728..2752)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2779..3351
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3548..3603
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4001..4018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4172..4760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4252..4271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4931..5722,5723..5791))
                     /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     5554..5573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5792..5896)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5964..5982
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6457..6512
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6709..7281
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7627..7646)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7896..7914
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9536..9545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagggatcct
      601 ctagtcagct gacgcgttac caccggtgcc accatgggaa gcgagctgat taaggagaac
      661 atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
      721 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
      781 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
      841 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
      901 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
      961 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
     1021 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
     1081 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
     1141 catctgatcg caaacgccaa gaccacatat agatccaaga aacccgctaa gaacctcaag
     1201 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
     1261 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
     1321 gggcacaagc ttaattccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac
     1381 ggtaccgcgg gcccgggatc caccggatct agaggaggaa gcggagctac taacttcagc
     1441 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatgtctga atatattcgg
     1501 gtaaccgaag atgagaacga tgagcccatt gaaataccat cggaagacga tgggacggtg
     1561 ctgctctcca cggttacagc ccagtttcca ggggcgtgtg ggcttcgcta caggaatcca
     1621 gtgtctcagt gtatgagagg tgtccggctg gtagaaggaa ttctgcatgc cccagatgct
     1681 ggctggggaa atctggtgta tgttgtcaac tatccaaaag ataacaaaag aaaaatggat
     1741 gagacagatg cttcatcagc agtgaaagtg aaaagagcag tccagaaaac atccgattta
     1801 atagtgttgg gtctcccatg gaaaacaacc gaacaggacc tgaaagagta ttttagtacc
     1861 tttggagaag ttcttatggt gcaggtcaag aaagatctta agactggtca ttcaaagggg
     1921 tttggctttg ttcgttttac ggaatatgaa acacaagtga aagtaatgtc acagcgacat
     1981 atgatagatg gacgatggtg tgactgcaaa cttcctaatt ctaagcaaag ccaagatgag
     2041 cctttgagaa gcagaaaagt gtttgtgggg cgctgtacag aggacatgac tgaggatgag
     2101 ctgcgggagt tcttctctca gtacggggat gtgatggatg tcttcatccc caagccattc
     2161 agggcctttg cctttgttac atttgcagat gatcagattg cgcagtctct ttgtggagag
     2221 gacttgatca ttaaaggaat cagcgttcat atatccaatg ccgaacctaa gcataacagt
     2281 aatcggcaat ttgagcgatc cggcagattt ggagggaacc ctggaggatt cggcaaccag
     2341 gggggctttg gaaattcaag aggagggggt gcgggattcg gtaacaacca ggggtccaat
     2401 ttcggtgggg ggttcaactt tggggcgttt tctttcaatc ccgctatgat ggctgcggct
     2461 caagcggctc tgcaaagtag ctggggcatg atgggtatgc tcgcgtcaca acagaatcag
     2521 agtggaccaa gtggaaataa tcagaatcaa gggaattttc aacgagaacc taaccaagca
     2581 ttcggatctg gtaataatag ctactccggg agtaactccg gagcagcttt cgggtggggc
     2641 tccgcgtcaa acgctggcag cggctctgga ttcaacgggg gattcggaag cagtttcgat
     2701 tcaaaaagca gtggctgggg gttctagacc cagctttctt gtacaaagtg gttgatcctc
     2761 tagactgaga acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt
     2821 gtaaaattca tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga
     2881 tgtcccttgt atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt
     2941 tgacaaccat tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt
     3001 agcttgcatt tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac
     3061 tttctctaat cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag
     3121 ttttagagaa caattgttat aattaaatga taaggtagaa tatttctgca tataaattct
     3181 ggctggcgtg gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct
     3241 ttctctttat ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc
     3301 aaaccgggcc cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc
     3361 aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag
     3421 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     3481 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     3541 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     3601 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     3661 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     3721 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     3781 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     3841 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     3901 atagctgtcc ctcttctctt atggagatcc ctcgactgca ttaatgaatc ggccaacgcg
     3961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5521 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     5581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5821 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5881 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     5941 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcactcga
     6001 cgcggccagc ttggtcgagc tggatacttc ccgtccgcca gggggacatg ccggcgatgc
     6061 tgaaggtcgc gcgcattccc gatgaagagg ccggttaccg cctgttgacc tggtgggacg
     6121 ggcagggcgc cgagttcata agagaagagg gacagctatg actgggagta gtcaggagag
     6181 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     6241 taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa
     6301 ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc aacctttgtt
     6361 catggcagcc agcatatggg catatgttgc caaactctaa accaaatact cattctgatg
     6421 ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca
     6481 cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt
     6541 catgatgtcc ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag
     6601 ggcattggcc acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaattc
     6661 tttgccaaaa tgatgagaca gcacaacaac cagcacgttg cccaggagct gtaggaaaga
     6721 gaagaaggca tgaacatggt tagcagaggg gcccggtttg gactcagagt attttatcct
     6781 catctcaaac agtgtatatc attgtaacca taaagagaaa ggcaggatga tgaccagggt
     6841 gtagttgttt ctaccaataa gaatatttcc acgccagcca gaatttatat gcagaaatat
     6901 tctaccttat catttaatta taacaattgt tctctaaaac tgtgctgaag tacaatataa
     6961 tataccctga ttgccttgaa aaaaaagtga ttagagaaag tacttacaat ctgacaaata
     7021 aacaaaagtg aatttaaaaa ttcgttacaa atgcaagcta aagtttaacg aaaaagttac
     7081 agaaaatgaa aagaaaataa gaggagacaa tggttgtcaa cagagtagaa agtgaaagaa
     7141 acaaaattat catgagggtc catggtgata caagggacat cttcccattc taaacaacac
     7201 cctgaaaact ttgccccctc catataacat gaattttaca atagcgaaaa agaaagaaca
     7261 atcaagggtc cccaaactca ccctgaagtt ctcagtctag agtcgagagg atcaatcact
     7321 ttgtacaaga aagctgggtc tagatagctt acttgtacag ctcgtccatg ccgccggtgg
     7381 agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     7441 tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct
     7501 tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct
     7561 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc
     7621 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     7681 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     7741 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt
     7801 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg
     7861 acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     7921 gggtgccctc gtaggggcgg ccctgaggaa aaaaaaggaa agggcatgct cgccatgtgc
     7981 aagatacagt gttgaatgtg gtgcaaaata taaagatgtg gaagaaaaag ccactgaaaa
     8041 taatatgagg aaataaaaaa ctagagtaaa ttatcagtgg aacaatttct attatacaag
     8101 acactacacc catacattct cctaatgctc atgtaagaat ttctccaaaa ataccttcca
     8161 gcactacaaa ctagaaaaaa aaagagacat ggacaccaaa ttaagttctt aatagtgaag
     8221 aacaaaagaa ctaccttgcc tgctccagtg gatccagcaa ccgccaacaa ctgtcctctt
     8281 tctatcttga aattaatatc tttcaggaca ggagtaccaa gaagtgagaa attactgaag
     8341 aagaggctgt catcaccatt agaagttttt ctattgttat tgttttgttt tgctttctca
     8401 aataattccc caaatccctg ttaaaaaaaa acacacacac acacacacac acacacatca
     8461 aaaataaaag atgagtttgt cagatatttt caatagatgc ttgttttatg taccatactc
     8521 acctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     8581 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctggttccg
     8641 gtgcctccac tttcatcttc acctttagtg ataccggctg ccgtcacgaa ctccttcagg
     8701 accatgtgat cgcgcttctc gttcgggtct ttgctcagcg tggtctgggt gctcagatag
     8761 tggttgtccg gcaggcgcac cgggccgtcg ccgatcgggg tgttctgctg tttgtggtcg
     8821 gcttcctgct ggctgccgtc ctcaacgttg tggcggatct tgaagttagc cttgatgccg
     8881 ttcttctgct tgtcggcttc gatacggacg ttgtggctgt tgtggttgta ctccagcttg
     8941 tgacccagga tgttgccgtc ctccttgaag tcgttgccct tcagctcgat gcggttcacc
     9001 agggtgtcgc cctcgaactt cacctccgcg cgggtcttgt aggtgccgtc gtccttgaac
     9061 tcgatggtgc gctcctggac gtagccttcc ggcatggcgg acttgaagaa gtcgtgacgc
     9121 ttcatgtggt ccgggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     9181 gtcggccacg gcaccggcag cttgccggtg gtgcagatga acttcagggt cagcttgccc
     9241 tcggtggcat cgccctcgcc ctcgccacgc acgctgaact tgtggccgtt tacgtcgccg
     9301 tccagctcga ctttgatagg cacagtacct gtaaatagtt cctctccttt actggtacca
     9361 tcatcaggga atgagcttac atttctcctc ttcagcatct ggtcatcctt cttagctttc
     9421 ctcagctcca cattgacctc tattctgcga cgcctcattt ctgtactgtc ttttcccttg
     9481 ttcttgaatc tgtgaagacg ggcagctggt gtattagcat tctcgttggt ggatcccatg
     9541 gtggcaccgg tgaattctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     9601 ggcctcccac cgtacacgcc acctcgacat actcgagta
//
