LOCUS       sequence-231210-        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: pHBS1399 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43
            G3...
FEATURES             Location/Qualifiers
     source          1..9667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             630..1331
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    1332..1397
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1422..1478
                     /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(2724..2748)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          2803..3375
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    3572..3627
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4025..4042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4196..4784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4276..4295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4955..5746,5747..5815))
                     /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     5578..5597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5816..5920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5988..6006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    6481..6536
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          6733..7305
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7651..7670)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7920..7938
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9560..9569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagg gatcctctag
      601 tcagctgacg cgttaccacc ggtgccacca tgggaagcga gctgattaag gagaacatgc
      661 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
      721 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
      781 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
      841 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
      901 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
      961 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     1021 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     1081 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     1141 tgatcgcaaa cgccaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     1201 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     1261 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     1321 acaagcttaa ttccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     1381 ccgcgggccc gggatccacc ggatctagag gaggaagcgg agctactaac ttcagcctgc
     1441 tgaagcaggc tggagacgtg gaggagaacc ctggacctat gtctgaatat attcgggtaa
     1501 ccgaagatga gaacgatgag cccattgaaa taccatcgga agacgatggg acggtgctgc
     1561 tctccacggt tacagcccag tttccagggg cgtgtgggct tcgctacagg aatccagtgt
     1621 ctcagtgtat gagaggtgtc cggctggtag aaggaattct gcatgcccca gatgctggct
     1681 ggggaaatct ggtgtatgtt gtcaactatc caaaagataa caaaagaaaa atggatgaga
     1741 cagatgcttc atcagcagtg aaagtgaaaa gagcagtcca gaaaacatcc gatttaatag
     1801 tgttgggtct cccatggaaa acaaccgaac aggacctgaa agagtatttt agtacctttg
     1861 gagaagttct tatggtgcag gtcaagaaag atcttaagac tggtcattca aaggggtttg
     1921 gctttgttcg ttttacggaa tatgaaacac aagtgaaagt aatgtcacag cgacatatga
     1981 tagatggacg atggtgtgac tgcaaacttc ctaattctaa gcaaagccaa gatgagcctt
     2041 tgagaagcag aaaagtgttt gtggggcgct gtacagagga catgactgag gatgagctgc
     2101 gggagttctt ctctcagtac ggggatgtga tggatgtctt catccccaag ccattcaggg
     2161 cctttgcctt tgttacattt gcagatgatc agattgcgca gtctctttgt ggagaggact
     2221 tgatcattaa aggaatcagc gttcatatat ccaatgccga acctaagcac aatagcaata
     2281 gacagttaga aagaagtgga agatttggtg gtaatccagg tggctttggg aatcagggtg
     2341 gatttggtaa tagcagaggg ggtggagctg gtttgggaaa caatcaaggt agtaatatgg
     2401 gtggtgggat gaactttggt gcgttcagca ttaatccagc catgatggct gccgcccagg
     2461 cagcactaca gagcagttgg ggtatgatgg gcatgttagc cagccagcag aaccagtcag
     2521 gcccatcggg taataaccaa aaccaaggca acatgcagag ggagccaaac caggccttct
     2581 ggtctggaaa taactcttat agtggctcta attctggtgc agcaattggt gggggatcag
     2641 catccaatgc agggtcgggc agtggtttta atggaggctt tggctcaagc atggattcta
     2701 agtcttctgg ctggggaatg tagacccagc tttcttgtac aaagtggttg atcctctact
     2761 agtaagcttg catgcgatat ctctagagct gagaacttca gggtgagttt ggggaccctt
     2821 gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggaggggg caaagttttc
     2881 agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc atgataattt
     2941 tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt cttttcattt
     3001 tctgtaactt tttcgttaaa ctttagcttg catttgtaac gaatttttaa attcactttt
     3061 gtttatttgt cagattgtaa gtactttctc taatcacttt tttttcaagg caatcagggt
     3121 atattatatt gtacttcagc acagttttag agaacaattg ttataattaa atgataaggt
     3181 agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg tagaaacaac
     3241 tacaccctgg tcatcatcct gcctttctct ttatggttac aatgatatac actgtttgag
     3301 atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc
     3361 ttctctttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc
     3421 aaagaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     3481 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     3541 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     3601 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3661 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     3721 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     3781 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     3841 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     3901 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3961 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4021 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     5581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5821 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5881 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5941 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6001 cgaggccctt tcgtcttcac tcgacgcggc cagcttggtc gagctggata cttcccgtcc
     6061 gccaggggga catgccggcg atgctgaagg tcgcgcgcat tcccgatgaa gaggccggtt
     6121 accgcctgtt gacctggtgg gacgggcagg gcgccgagtt cataagagaa gagggacagc
     6181 tatgactggg agtagtcagg agaggaggaa aaatctggct agtaaaacat gtaaggaaaa
     6241 ttttagggat gttaaagaaa aaaataacac aaaacaaaat ataaaaaaaa tctaacctca
     6301 agtcaaggct tttctatgga ataaggaatg gacagcaggg ggctgtttca tatactgatg
     6361 acctctttat agccaacctt tgttcatggc agccagcata tgggcatatg ttgccaaact
     6421 ctaaaccaaa tactcattct gatgttttaa atgatttgcc ctcccatatg tccttccgag
     6481 tgagagacac aaaaaattcc aacacactat tgcaatgaaa ataaatttcc tttattagcc
     6541 agaagtcaga tgctcaaggg gcttcatgat gtccccataa tttttggcag agggaaaaag
     6601 atctcagtgg tatttgtgag ccagggcatt ggccacacca gccaccacct tctgataggc
     6661 agcctgcacc tgaggagtga attctttgcc aaaatgatga gacagcacaa caaccagcac
     6721 gttgcccagg agctgtagga aagagaagaa ggcatgaaca tggttagcag aggggcccgg
     6781 tttggactca gagtatttta tcctcatctc aaacagtgta tatcattgta accataaaga
     6841 gaaaggcagg atgatgacca gggtgtagtt gtttctacca ataagaatat ttccacgcca
     6901 gccagaattt atatgcagaa atattctacc ttatcattta attataacaa ttgttctcta
     6961 aaactgtgct gaagtacaat ataatatacc ctgattgcct tgaaaaaaaa gtgattagag
     7021 aaagtactta caatctgaca aataaacaaa agtgaattta aaaattcgtt acaaatgcaa
     7081 gctaaagttt aacgaaaaag ttacagaaaa tgaaaagaaa ataagaggag acaatggttg
     7141 tcaacagagt agaaagtgaa agaaacaaaa ttatcatgag ggtccatggt gatacaaggg
     7201 acatcttccc attctaaaca acaccctgaa aactttgccc cctccatata acatgaattt
     7261 tacaatagcg aaaaagaaag aacaatcaag ggtccccaaa ctcaccctga agttctcagt
     7321 ctagagtcga gaggatcaat cactttgtac aagaaagctg ggtctagata gcttacttgt
     7381 acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga
     7441 tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg
     7501 gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc
     7561 cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca
     7621 tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg
     7681 ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc agggaggagt
     7741 cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc
     7801 cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct
     7861 tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg gggccaccct
     7921 tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctga ggaaaaaaaa
     7981 ggaaagggca tgctcgccat gtgcaagata cagtgttgaa tgtggtgcaa aatataaaga
     8041 tgtggaagaa aaagccactg aaaataatat gaggaaataa aaaactagag taaattatca
     8101 gtggaacaat ttctattata caagacacta cacccataca ttctcctaat gctcatgtaa
     8161 gaatttctcc aaaaatacct tccagcacta caaactagaa aaaaaaagag acatggacac
     8221 caaattaagt tcttaatagt gaagaacaaa agaactacct tgcctgctcc agtggatcca
     8281 gcaaccgcca acaactgtcc tctttctatc ttgaaattaa tatctttcag gacaggagta
     8341 ccaagaagtg agaaattact gaagaagagg ctgtcatcac cattagaagt ttttctattg
     8401 ttattgtttt gttttgcttt ctcaaataat tccccaaatc cctgttaaaa aaaaacacac
     8461 acacacacac acacacacac atcaaaaata aaagatgagt ttgtcagata ttttcaatag
     8521 atgcttgttt tatgtaccat actcacctcg ccctcgccct cgatctcgaa ctcgtggccg
     8581 ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgtta
     8641 tcctcctcgc ccttgctggt tccggtgcct ccactttcat cttcaccttt agtgataccg
     8701 gctgccgtca cgaactcctt caggaccatg tgatcgcgct tctcgttcgg gtctttgctc
     8761 agcgtggtct gggtgctcag atagtggttg tccggcaggc gcaccgggcc gtcgccgatc
     8821 ggggtgttct gctgtttgtg gtcggcttcc tgctggctgc cgtcctcaac gttgtggcgg
     8881 atcttgaagt tagccttgat gccgttcttc tgcttgtcgg cttcgatacg gacgttgtgg
     8941 ctgttgtggt tgtactccag cttgtgaccc aggatgttgc cgtcctcctt gaagtcgttg
     9001 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc cgcgcgggtc
     9061 ttgtaggtgc cgtcgtcctt gaactcgatg gtgcgctcct ggacgtagcc ttccggcatg
     9121 gcggacttga agaagtcgtg acgcttcatg tggtccgggt agcggctgaa gcactgcacg
     9181 ccgtaggtca gggtggtcac gagggtcggc cacggcaccg gcagcttgcc ggtggtgcag
     9241 atgaacttca gggtcagctt gccctcggtg gcatcgccct cgccctcgcc acgcacgctg
     9301 aacttgtggc cgtttacgtc gccgtccagc tcgactttga taggcacagt acctgtaaat
     9361 agttcctctc ctttactggt accatcatca gggaatgagc ttacatttct cctcttcagc
     9421 atctggtcat ccttcttagc tttcctcagc tccacattga cctctattct gcgacgcctc
     9481 atttctgtac tgtcttttcc cttgttcttg aatctgtgaa gacgggcagc tggtgtatta
     9541 gcattctcgt tggtggatcc catggtggca ccggtgaatt ctccaggcga tctgacggtt
     9601 cactaaacga gctctgctta tataggcctc ccaccgtaca cgccacctcg acatactcga
     9661 gtagtta
//
