LOCUS       sequence-261767-        9072 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 9072)
  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 9072)
  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: pHBS1501 [IBB-GFP-mCherry3E]-[BFP-TDP43 deltaRRM]
FEATURES             Location/Qualifiers
     source          1..9072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    63..118
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(516..533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(687..1275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(767..786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1446..2237,2238..2306))
                     /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     2069..2088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2307..2411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2479..2497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    2972..3027
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          3224..3796
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4142..4161)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4411..4429
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6051..6060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        6213..6516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6517..6720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6671..6691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6782..6791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6788..7489
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(8287..8311)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          8366..8938
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
       61 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
      121 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
      181 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
      241 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
      301 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
      361 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
      421 tgtccctctt ctcttatgga gatccctcga ctgcattaat gaatcggcca acgcgcgggg
      481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     1921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2041 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     2101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2401 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2461 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca ctcgacgcgg
     2521 ccagcttggt cgagctggat acttcccgtc cgccaggggg acatgccggc gatgctgaag
     2581 gtcgcgcgca ttcccgatga agaggccggt taccgcctgt tgacctggtg ggacgggcag
     2641 ggcgccgagt tcataagaga agagggacag ctatgactgg gagtagtcag gagaggagga
     2701 aaaatctggc tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca
     2761 caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat
     2821 ggacagcagg gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg
     2881 cagccagcat atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta
     2941 aatgatttgc cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta
     3001 ttgcaatgaa aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga
     3061 tgtccccata atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat
     3121 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc
     3181 caaaatgatg agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga
     3241 aggcatgaac atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct
     3301 caaacagtgt atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt
     3361 tgtttctacc aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac
     3421 cttatcattt aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac
     3481 cctgattgcc ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa
     3541 aagtgaattt aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa
     3601 atgaaaagaa aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa
     3661 attatcatga gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga
     3721 aaactttgcc ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa
     3781 gggtccccaa actcaccctg aagttctcag tctagagtcg agaggatcaa tcactttgta
     3841 caagaaagct gggtctagat agcttacttg tacagctcgt ccatgccgcc ggtggagtgg
     3901 cggccctcgg cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg
     3961 tccaacttga tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag
     4021 gtggtcttga cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc
     4081 tcgcccttca gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg
     4141 gtcttcttct gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg
     4201 tagatgaact cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc
     4261 tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg
     4321 gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg
     4381 atgtcccagg cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg
     4441 ccctcgtagg ggcggccctg aggaaaaaaa aggaaagggc atgctcgcca tgtgcaagat
     4501 acagtgttga atgtggtgca aaatataaag atgtggaaga aaaagccact gaaaataata
     4561 tgaggaaata aaaaactaga gtaaattatc agtggaacaa tttctattat acaagacact
     4621 acacccatac attctcctaa tgctcatgta agaatttctc caaaaatacc ttccagcact
     4681 acaaactaga aaaaaaaaga gacatggaca ccaaattaag ttcttaatag tgaagaacaa
     4741 aagaactacc ttgcctgctc cagtggatcc agcaaccgcc aacaactgtc ctctttctat
     4801 cttgaaatta atatctttca ggacaggagt accaagaagt gagaaattac tgaagaagag
     4861 gctgtcatca ccattagaag tttttctatt gttattgttt tgttttgctt tctcaaataa
     4921 ttccccaaat ccctgttaaa aaaaaacaca cacacacaca cacacacaca catcaaaaat
     4981 aaaagatgag tttgtcagat attttcaata gatgcttgtt ttatgtacca tactcacctc
     5041 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     5101 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctgg ttccggtgcc
     5161 tccactttca tcttcacctt tagtgatacc ggctgccgtc acgaactcct tcaggaccat
     5221 gtgatcgcgc ttctcgttcg ggtctttgct cagcgtggtc tgggtgctca gatagtggtt
     5281 gtccggcagg cgcaccgggc cgtcgccgat cggggtgttc tgctgtttgt ggtcggcttc
     5341 ctgctggctg ccgtcctcaa cgttgtggcg gatcttgaag ttagccttga tgccgttctt
     5401 ctgcttgtcg gcttcgatac ggacgttgtg gctgttgtgg ttgtactcca gcttgtgacc
     5461 caggatgttg ccgtcctcct tgaagtcgtt gcccttcagc tcgatgcggt tcaccagggt
     5521 gtcgccctcg aacttcacct ccgcgcgggt cttgtaggtg ccgtcgtcct tgaactcgat
     5581 ggtgcgctcc tggacgtagc cttccggcat ggcggacttg aagaagtcgt gacgcttcat
     5641 gtggtccggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtcgg
     5701 ccacggcacc ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccctcggt
     5761 ggcatcgccc tcgccctcgc cacgcacgct gaacttgtgg ccgtttacgt cgccgtccag
     5821 ctcgactttg ataggcacag tacctgtaaa tagttcctct cctttactgg taccatcatc
     5881 agggaatgag cttacatttc tcctcttcag catctggtca tccttcttag ctttcctcag
     5941 ctccacattg acctctattc tgcgacgcct catttctgta ctgtcttttc ccttgttctt
     6001 gaatctgtga agacgggcag ctggtgtatt agcattctcg ttggtggatc ccatggtggc
     6061 accggtgaat tctccaggcg atctgacggt tcactaaacg agctctgctt atataggcct
     6121 cccaccgtac acgccacctc gacatactcg agtagttatt aatagtaatc aattacgggg
     6181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     6241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     6421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     6481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     6541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     6601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     6661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     6721 ggtttagtga accgtcagat ccgctaggga tcctctagtc agctgacgcg ttaccaccgg
     6781 tgccaccatg ggaagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg
     6841 caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     6901 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
     6961 cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc
     7021 cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     7081 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     7141 caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac
     7201 actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag
     7261 aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac
     7321 atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta
     7381 cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc
     7441 agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatt ccggaatgtc
     7501 tgaatatatt cgggtaaccg aagatgagaa cgatgagccc attgaaatac catcggaaga
     7561 cgatgggacg gtgctgctct ccacggttac agcccagttt ccaggggcgt gtgggcttcg
     7621 ctacaggaat ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag gaattctgca
     7681 tgccccagat gctggctggg gaaatctggt gtatgttgtc aactatccaa aagataacaa
     7741 aagaaaaatg gatgagacag atgcttcatc agcagtgaaa gtgaaaagag cagtccagaa
     7801 aacatcctcc aatgccgaac ctaagcacaa tagcaataga cagttagaaa gaagtggaag
     7861 atttggtggt aatccaggtg gctttgggaa tcagggtgga tttggtaata gcagaggggg
     7921 tggagctggt ttgggaaaca atcaaggtag taatatgggt ggtgggatga actttggtgc
     7981 gttcagcatt aatccagcca tgatggctgc cgcccaggca gcactacaga gcagttgggg
     8041 tatgatgggc atgttagcca gccagcagaa ccagtcaggc ccatcgggta ataaccaaaa
     8101 ccaaggcaac atgcagaggg agccaaacca ggccttcggt tctggaaata actcttatag
     8161 tggctctaat tctggtgcag caattggttg gggatcagca tccaatgcag ggtcgggcag
     8221 tggttttaat ggaggctttg gctcaagcat ggattctaag tcttctggct ggggaatgta
     8281 gactagaccc agctttcttg tacaaagtgg ttgatcctct actagtaagc ttgcatgcga
     8341 tatctctaga gctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt
     8401 cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat
     8461 gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct
     8521 actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt
     8581 aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg
     8641 taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc
     8701 agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat
     8761 aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat
     8821 cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc
     8881 tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct
     8941 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcactcctca
     9001 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     9061 ccactgagat ct
//
