LOCUS       sequence-203695-        9187 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 9187)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 9187)
  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"
COMMENT     Sequence Label: pHBS1109 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43  ∆...
FEATURES             Location/Qualifiers
     source          1..9187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    325..380
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     intron          577..1149
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    1179..1203
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1487..1506)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1756..1774
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3396..3405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        3562..3865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3866..4069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4020..4040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4131..4140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4137..4838
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    4839..4904
                     /label=MCS
                     /note="multiple cloning site"
     CDS             4929..4985
                     /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(5775..5799)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          5853..6425
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6622..6677
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7068..7085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7239..7827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7319..7338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7998..8789,8790..8858))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8621..8640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8859..8963)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9031..9049
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgagttcata agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat
       61 ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa
      121 caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca
      181 gcagggggct gtttcatata ctgatgacct ctttatagcc acctttgttc atggcagcca
      241 gcatatggca tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt
      301 tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttcccacaca ctattgcaat
      361 gaaaataaat ttcctttatt acccagaagt cagatgctca aggggcttca tgatgtcccc
      421 ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg cattggccac
      481 accagccacc accttctgat aggcagcctg cacctgagga gtgaattctt tgccaaaatg
      541 atgagacagc acaacaacca gcacgttgcc caggagctgt aggaaaaaga agaaggcatg
      601 aacatggtta gcagaggggc ccggtttgga ctcagagtat tttatcctca tctcaaacag
      661 tgtatatcat tgtaaccata aagagaaagg caggatgatt accaggatgt agttgtttct
      721 accaataaga atatttccac gccagccaga atttatatgc agaaatattc taccttatca
      781 tttaattata acaattgttc tctaaaactg tgctgaagta caatataatt accctgattg
      841 ccttgaaaaa aaagtgatta gagaaagtac ttacaatctg acaaataaac gaaagtgaat
      901 ttaaaaattc gttacaaatg caagctaaag tttaacgaaa aaagttacag aaaatgaaaa
      961 gaaaataaga ggagacaatg gttgtcaaca gagtagaaag tgaaagaaac aaaattatca
     1021 tgagggtcca tggtgataca agggacatct tcccattcta aacaacaccc tgaaaacttt
     1081 gccccctcca tataacatga atttttcaat agcgaaaaag aaagaacaat caagggtccc
     1141 caaactcacc ctgaagttct cagtctagag gaggatcaac cactttgtac aagaaagctg
     1201 aagctagttt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     1261 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     1321 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     1381 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     1441 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     1501 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     1561 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     1621 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     1681 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     1741 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     1801 ccctgaggaa aaaaaaggaa agggcatgct cgccatgtgc aagatacagt gttgaatgtg
     1861 gtgcaaaata taaagatgtg gaagaaaaag ccactgaaaa taatatgagg aaataaaaaa
     1921 ctagagtaaa ttatcagtgg aacaatttct attatacaag acactacacc catacattct
     1981 cctaatgctc atgtaagaat ttctccaaaa ataccttcca gcactacaaa ctagaaaaaa
     2041 aaagagacat ggacaccaaa ttaagttctt aatagtgaag aacaaaagaa ctaccttgcc
     2101 tgctccagtg gatccagcaa ccgccaacaa ctgtcctctt tctatcttga aattaatatc
     2161 tttcaggaca ggagtaccaa gaagtgagaa attactgaag aagaggctgt catcaccatt
     2221 agaagttttt ctattgttat tgttttgttt tgctttctca aattattccc caaatccctg
     2281 ttaaaaaaaa acacacacac acacacacac acacacatca aaaataaaag atgagtttgt
     2341 cagatatttt caatagatgc ttgttttatg taccatactc acctcgccct cgccctcgat
     2401 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     2461 gatgatggcc atgttatcct cctcgccctt gctggttccg gtgcctccac tttcatcttc
     2521 acctttagtg ataccggctg ccgtcacgaa ctccttcagg accatgtgat cgcgcttctc
     2581 gttcgggtct ttgctcagcg tggtctgggt gctcagatag tggttgtccg gcaggcgcac
     2641 cgggccgtcg ccgatcgggg tgttctgctg tttgtggtcg gcttcctgct ggctgccgtc
     2701 ctcaacgttg tggcggatct tgaagttagc cttgatgccg ttcttctgct tgtcggcttc
     2761 gatacggacg ttgtggctgt tgtggttgta ctccagcttg tgacccagga tgttgccgtc
     2821 ctccttgaag tcgttgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2881 cacctccgcg cgggtcttgt aggtgccgtc gtccttgaac tcgatggtgc gctcctggac
     2941 gtagccttcc ggcatggcgg acttgaagaa gtcgtgacgc ttcatgtggt ccgggtagcg
     3001 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtcggccacg gcaccggcag
     3061 cttgccggtg gtgcagatga acttcagggt cagcttgccc tcggtggcat cgccctcgcc
     3121 ctcgccacgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga ctttgatagg
     3181 cacagtacct gtaaatagtt cctctccttt actggtacca tcatcaggga atgagcttac
     3241 atttctcctc ttcagcatct ggtcatcctt cttagctttc ctcagctcca cattgacctc
     3301 tattctgcga cgcctcattt ctgtactgtc ttttcccttg ttcttgaatc tgtgaagacg
     3361 ggcagctggt gtattagcat tctcgttggt ggatcccatg gtggcaccgg ccggtgaatt
     3421 ctccaggcga tctgacggtt cactaaacga gctctgctta tataggcctc ccaccgtaca
     3481 cgccacctcg acatactcga gtagttatta atagtaatca attacggggt cattagttca
     3541 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3601 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3661 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3721 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     3781 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     3841 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     3901 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3961 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4021 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     4081 ccgtcagatc cgctagggat cctctagtca gctgacgcgt taccaccggt gccaccatgg
     4141 gaagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     4201 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     4261 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     4321 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     4381 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     4441 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     4501 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     4561 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     4621 ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca tatagatcca
     4681 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     4741 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     4801 actgcgacct ccctagcaaa ctggggcaca agcttaattc cggactcaga tctcgagctc
     4861 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagaggag
     4921 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     4981 gacctatgtc tgaatatatt cgggtaaccg aagatgagaa cgatgagccc attgaaatac
     5041 catcggaaga cgatgggacg gtgctgctct ccacggttac agcccagttt ccaggggcgt
     5101 gtgggcttcg ctacaggaat ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag
     5161 gaattctgca tgccccagat gctggctggg gaaatctggt gtatgttgtc aactatccaa
     5221 aagataacaa aagaaaaatg gatgagacag atgcttcatc agcagtgaaa gtgaaaagag
     5281 cagtccagaa aacatccgat ttaatagtgt tgggtctccc atggaaaaca accgaacagg
     5341 acctgaaaga gtattttagt acctttggag aagttcttat ggtgcaggtc aagaaagatc
     5401 ttaagactgg tcattcaaag gggtttggct ttgttcgttt tacggaatat gaaacacaag
     5461 tgaaagtaat gtcacagcga catatgatag atggacgatg gtgtgactgc aaacttccta
     5521 attctaagca aagccaagat gagcctttga gaagcagaaa agtgtttgtg gggcgctgta
     5581 cagaggacat gactgaggat gagctgcggg agttcttctc tcagtacggg gatgtgatgg
     5641 atgtcttcat ccccaagcca ttcagggcct ttgcctttgt tacatttgca gatgatcaga
     5701 ttgcgcagtc tctttgtgga gaggacttga tcattaaagg aatcagcgtt catatatcca
     5761 atgccgaacc ttagacccag ctttcttgta caaagtggtt gatcctctac tagtaagctt
     5821 gcatgcgata tctctagact gagaacttca gggtgagttt ggggaccctt gattgttctt
     5881 tctttttcgc tattgaaaaa ttcatgttat atggaggggg caaagttttc agggtgttgt
     5941 ttagaatggg aagatgtccc ttgtatcacc atggaccctc atgataattt tgtttctttc
     6001 actttctact ctgttgacaa ccattgtctc ctcttatttt cttttcattt tctgtaactt
     6061 ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt cgtttatttg
     6121 tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg taattatatt
     6181 gtacttcagc acagttttag agaacaattg ttataattaa atgataaggt agaatatttc
     6241 tgcatataaa ttctggctgg cgtggaaata ttcttattgg tagaaacaac tacatcctgg
     6301 taatcatcct gcctttctct ttatggttac aatgatatac actgtttgag atgaggataa
     6361 aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc ttctttttcc
     6421 tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaagaattca
     6481 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     6541 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     6601 ttgagcatct gacttctggg taataaagga aatttatttt cattgcaata gtgtgtggga
     6661 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     6721 atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca tgaacaaagg
     6781 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     6841 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     6901 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     6961 ctactcccag tcatagctgt ccctcttctc ttatgaactc gactgcatta atgaatcggc
     7021 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7081 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7141 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7201 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7261 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7321 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7381 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     7441 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7501 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7561 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7621 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7681 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7741 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7801 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7861 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7921 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7981 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8041 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8101 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8161 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8221 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8281 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8341 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8401 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8461 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8521 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8581 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8641 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8701 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8761 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8821 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8881 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8941 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     9001 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     9061 cactcgacgc tggtcgagct gatacttccc gtccgccagg ggacatgccg gcgatgctga
     9121 aggtcgcgcg cattcccgat gaagaggccg gttaccgcct gttgacctgg tgggacgggc
     9181 agggcgc
//
