LOCUS       sequence-261752-        9498 bp    DNA     circular SYN 15-APR-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 Apr 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: pHBS1507 [IBB-GFP-mCherry 3E]-[BFP-TDP43 VLIM-S]
FEATURES             Location/Qualifiers
     source          1..9498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(880..899)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1149..1167
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2789..2798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        2951..3254
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3255..3458
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3409..3429
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3520..3529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3526..4227
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     protein_bind    complement(5479..5503)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          5530..6102
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6299..6354
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6752..6769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6923..7511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7003..7022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7682..8473,8474..8542))
                     /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     8305..8324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8543..8647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8715..8733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    9208..9263
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          join(9460..9498,1..534)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 ggggcccggt ttggactcag agtattttat cctcatctca aacagtgtat atcattgtaa
       61 ccataaagag aaaggcagga tgatgaccag ggtgtagttg tttctaccaa taagaatatt
      121 tccacgccag ccagaattta tatgcagaaa tattctacct tatcatttaa ttataacaat
      181 tgttctctaa aactgtgctg aagtacaata taatataccc tgattgcctt gaaaaaaaag
      241 tgattagaga aagtacttac aatctgacaa ataaacaaaa gtgaatttaa aaattcgtta
      301 caaatgcaag ctaaagttta acgaaaaagt tacagaaaat gaaaagaaaa taagaggaga
      361 caatggttgt caacagagta gaaagtgaaa gaaacaaaat tatcatgagg gtccatggtg
      421 atacaaggga catcttccca ttctaaacaa caccctgaaa actttgcccc ctccatataa
      481 catgaatttt acaatagcga aaaagaaaga acaatcaagg gtccccaaac tcaccctgaa
      541 gttctcagtc tagagtcgag aggatcaatc actttgtaca agaaagctgg gtctagatag
      601 cttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact
      661 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt
      721 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt
      781 agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct
      841 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
      901 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
      961 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
     1021 acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca
     1081 cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg
     1141 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctgag
     1201 gaaaaaaaag gaaagggcat gctcgccatg tgcaagatac agtgttgaat gtggtgcaaa
     1261 atataaagat gtggaagaaa aagccactga aaataatatg aggaaataaa aaactagagt
     1321 aaattatcag tggaacaatt tctattatac aagacactac acccatacat tctcctaatg
     1381 ctcatgtaag aatttctcca aaaatacctt ccagcactac aaactagaaa aaaaaagaga
     1441 catggacacc aaattaagtt cttaatagtg aagaacaaaa gaactacctt gcctgctcca
     1501 gtggatccag caaccgccaa caactgtcct ctttctatct tgaaattaat atctttcagg
     1561 acaggagtac caagaagtga gaaattactg aagaagaggc tgtcatcacc attagaagtt
     1621 tttctattgt tattgttttg ttttgctttc tcaaataatt ccccaaatcc ctgttaaaaa
     1681 aaaacacaca cacacacaca cacacacaca tcaaaaataa aagatgagtt tgtcagatat
     1741 tttcaataga tgcttgtttt atgtaccata ctcacctcgc cctcgccctc gatctcgaac
     1801 tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg
     1861 gccatgttat cctcctcgcc cttgctggtt ccggtgcctc cactttcatc ttcaccttta
     1921 gtgataccgg ctgccgtcac gaactccttc aggaccatgt gatcgcgctt ctcgttcggg
     1981 tctttgctca gcgtggtctg ggtgctcaga tagtggttgt ccggcaggcg caccgggccg
     2041 tcgccgatcg gggtgttctg ctgtttgtgg tcggcttcct gctggctgcc gtcctcaacg
     2101 ttgtggcgga tcttgaagtt agccttgatg ccgttcttct gcttgtcggc ttcgatacgg
     2161 acgttgtggc tgttgtggtt gtactccagc ttgtgaccca ggatgttgcc gtcctccttg
     2221 aagtcgttgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcc
     2281 gcgcgggtct tgtaggtgcc gtcgtccttg aactcgatgg tgcgctcctg gacgtagcct
     2341 tccggcatgg cggacttgaa gaagtcgtga cgcttcatgt ggtccgggta gcggctgaag
     2401 cactgcacgc cgtaggtcag ggtggtcacg agggtcggcc acggcaccgg cagcttgccg
     2461 gtggtgcaga tgaacttcag ggtcagcttg ccctcggtgg catcgccctc gccctcgcca
     2521 cgcacgctga acttgtggcc gtttacgtcg ccgtccagct cgactttgat aggcacagta
     2581 cctgtaaata gttcctctcc tttactggta ccatcatcag ggaatgagct tacatttctc
     2641 ctcttcagca tctggtcatc cttcttagct ttcctcagct ccacattgac ctctattctg
     2701 cgacgcctca tttctgtact gtcttttccc ttgttcttga atctgtgaag acgggcagct
     2761 ggtgtattag cattctcgtt ggtggatccc atggtggcac cggtgaattc tccaggcgat
     2821 ctgacggttc actaaacgag ctctgcttat ataggcctcc caccgtacac gccacctcga
     2881 catactcgag tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2941 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3001 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3061 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3121 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3181 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3241 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     3301 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     3361 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     3421 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     3481 gctagggatc ctctagtcag ctgacgcgtt accaccggtg ccaccatggg aagcgagctg
     3541 attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc
     3601 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     3661 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac
     3721 ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc
     3781 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     3841 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     3901 aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc
     3961 gagacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc
     4021 gtgggcggga gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct
     4081 aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag
     4141 gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc
     4201 cctagcaaac tggggcacaa gcttaattcc ggaatgtctg aatatattcg ggtaaccgaa
     4261 gatgagaacg atgagcccat tgaaatacca tcggaagacg atgggacggt gctgctctcc
     4321 acggttacag cccagtttcc aggggcgtgt gggcttcgct acaggaatcc agtgtctcag
     4381 tgtatgagag gtgtccggct ggtagaagga attctgcatg ccccagatgc tggctgggga
     4441 aatctggtgt atgttgtcaa ctatccaaaa gataacaaaa gaaaaatgga tgagacagat
     4501 gcttcatcag cagtgaaagt gaaaagagca gtccagaaaa catccgattt aatagtgttg
     4561 ggtctcccat ggaaaacaac cgaacaggac ctgaaagagt attttagtac ctttggagaa
     4621 gttcttatgg tgcaggtcaa gaaagatctt aagactggtc attcaaaggg gtttggcttt
     4681 gttcgtttta cggaatatga aacacaagtg aaagtaatgt cacagcgaca tatgatagat
     4741 ggacgatggt gtgactgcaa acttcctaat tctaagcaaa gccaagatga gcctttgaga
     4801 agcagaaaag tgtttgtggg gcgctgtaca gaggacatga ctgaggatga gctgcgggag
     4861 ttcttctctc agtacgggga tgtgatggat gtcttcatcc ccaagccatt cagggccttt
     4921 gcctttgtta catttgcaga tgatcagatt gcgcagtctc tttgtggaga ggacttgatc
     4981 attaaaggaa tcagcgttca tatatccaat gccgaaccta agcacaacag taatcgacag
     5041 tccgagagaa gcgggcggtt cggtggtaat ccaggggggt tcggaaacca aggaggtttt
     5101 ggtaactcta ggggaggagg cgctggttcc ggtaataacc aaggcagcaa cagtgggggg
     5161 ggcagtaatt tcggtgcttt ttcaagtaac ccagcaatga tggccgccgc acaagcagct
     5221 ttgcaaagtt cttggggcat gatgggcatg ctggcgtcac agcaaaatca aagcggaccc
     5281 tcaggaaata atcaaaacca aggcaatagc cagagggagc ctaatcaggc cttcggatca
     5341 ggcaataatt catacagtgg ttcaaactcc ggggcagcct ctgggtgggg cagcgcaagt
     5401 aatgccggaa gcggtagtgg attcaacgga gggttcgggt caagtagcga tagtaaaagt
     5461 tccggttggg gctcttagac ccagctttct tgtacaaagt ggttgatcct ctagactgag
     5521 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
     5581 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
     5641 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
     5701 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
     5761 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
     5821 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
     5881 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
     5941 ggaaatattc ttattggtag aaacaactac accctggtca tcatcctgcc tttctcttta
     6001 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
     6061 ccctctgcta accatgttca tgccttcttc tctttcctac agctcctggg caacgtgctg
     6121 gttgttgtgc tgtctcatca ttttggcaaa gaattcactc ctcaggtgca ggctgcctat
     6181 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     6241 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     6301 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     6361 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     6421 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     6481 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     6541 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     6601 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     6661 cctcttctct tatggagatc cctcgactgc attaatgaat cggccaacgc gcggggagag
     6721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8281 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactcg acgcggccag
     8761 cttggtcgag ctggatactt cccgtccgcc agggggacat gccggcgatg ctgaaggtcg
     8821 cgcgcattcc cgatgaagag gccggttacc gcctgttgac ctggtgggac gggcagggcg
     8881 ccgagttcat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     8941 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     9001 acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac
     9061 agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc
     9121 cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg
     9181 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc
     9241 aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc
     9301 cccataattt ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca gggcattggc
     9361 cacaccagcc accaccttct gataggcagc ctgcacctga ggagtgaatt ctttgccaaa
     9421 atgatgagac agcacaacaa ccagcacgtt gcccaggagc tgtaggaaag agaagaaggc
     9481 atgaacatgg ttagcaga
//
