LOCUS       sequence-208308-        8367 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 splicing activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8367)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 8367)
  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: pHBS1333 (IBB-GFP-mCherry 3E splicer)-(BFP-P2A...
FEATURES             Location/Qualifiers
     source          1..8367
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(224..243)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     493..511
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2133..2142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        2295..2598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2599..2802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2753..2773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2864..2873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2870..3571
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    3572..3637
                     /label=MCS
                     /note="multiple cloning site"
     CDS             3662..3718
                     /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(4964..4988)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          5016..5588
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(5720..5739)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5785..5840
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5839..5858)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6238..6255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6409..6997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6489..6508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7168..7959,7960..8028))
                     /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     7791..7810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8029..8133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8201..8219
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
       61 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
      121 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
      181 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
      241 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
      301 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
      361 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
      421 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
      481 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
      541 tgaggaaaaa aaaggaaagg gcatgctcgc catgtgcaag atacagtgtt gaatgtggtg
      601 caaaatataa agatgtggaa gaaaaagcca ctgaaaataa tatgaggaaa taaaaaacta
      661 gagtaaatta tcagtggaac aatttctatt atacaagaca ctacacccat acattctcct
      721 aatgctcatg taagaatttc tccaaaaata ccttccagca ctacaaacta gaaaaaaaaa
      781 gagacatgga caccaaatta agttcttaat agtgaagaac aaaagaacta ccttgcctgc
      841 tccagtggat ccagcaaccg ccaacaactg tcctctttct atcttgaaat taatatcttt
      901 caggacagga gtaccaagaa gtgagaaatt actgaagaag aggctgtcat caccattaga
      961 agtttttcta ttgttattgt tttgttttgc tttctcaaat tattccccaa atccctgtta
     1021 aaaaaaaaca cacacacaca cacacacaca cacatcaaaa ataaaagatg agtttgtcag
     1081 atattttcaa tagatgcttg ttttatgtac catactcacc tcgccctcgc cctcgatctc
     1141 gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat
     1201 gatggccatg ttatcctcct cgcccttgct ggttccggtg cctccacttt catcttcacc
     1261 tttagtgata ccggctgccg tcacgaactc cttcaggacc atgtgatcgc gcttctcgtt
     1321 cgggtctttg ctcagcgtgg tctgggtgct cagatagtgg ttgtccggca ggcgcaccgg
     1381 gccgtcgccg atcggggtgt tctgctgttt gtggtcggct tcctgctggc tgccgtcctc
     1441 aacgttgtgg cggatcttga agttagcctt gatgccgttc ttctgcttgt cggcttcgat
     1501 acggacgttg tggctgttgt ggttgtactc cagcttgtga cccaggatgt tgccgtcctc
     1561 cttgaagtcg ttgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     1621 ctccgcgcgg gtcttgtagg tgccgtcgtc cttgaactcg atggtgcgct cctggacgta
     1681 gccttccggc atggcggact tgaagaagtc gtgacgcttc atgtggtccg ggtagcggct
     1741 gaagcactgc acgccgtagg tcagggtggt cacgagggtc ggccacggca ccggcagctt
     1801 gccggtggtg cagatgaact tcagggtcag cttgccctcg gtggcatcgc cctcgccctc
     1861 gccacgcacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgactt tgataggcac
     1921 agtacctgta aatagttcct ctcctttact ggtaccatca tcagggaatg agcttacatt
     1981 tctcctcttc agcatctggt catccttctt agctttcctc agctccacat tgacctctat
     2041 tctgcgacgc ctcatttctg tactgtcttt tcccttgttc ttgaatctgt gaagacgggc
     2101 agctggtgta ttagcattct cgttggtgga tcccatggtg gcaccggtga attctccagg
     2161 cgatctgacg gttcactaaa cgagctctgc ttatataggc ctcccaccgt acacgccacc
     2221 tcgacatact cgagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     2281 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2341 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2401 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2461 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2521 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     2581 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     2641 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2701 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2761 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     2821 atccgctagg gatcctctag tcagctgacg cgttaccacc ggtgccacca tgggaagcga
     2881 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca
     2941 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     3001 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct
     3061 ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc
     3121 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     3181 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     3241 ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt
     3301 caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa
     3361 gctcgtgggc gggagccatc tgatcgcaaa cgccaagacc acatatagat ccaagaaacc
     3421 cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa
     3481 ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga
     3541 cctccctagc aaactggggc acaagcttaa ttccggactc agatctcgag ctcaagcttc
     3601 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagag gaggaagcgg
     3661 agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctat
     3721 gtctgaatat attcgggtaa ccgaagatga gaacgatgag cccattgaaa taccatcgga
     3781 agacgatggg acggtgctgc tctccacggt tacagcccag tttccagggg cgtgtgggct
     3841 tcgctacagg aatccagtgt ctcagtgtat gagaggtgtc cggctggtag aaggaattct
     3901 gcatgcccca gatgctggct ggggaaatct ggtgtatgtt gtcaactatc caaaagataa
     3961 caaaagaaaa atggatgaga cagatgcttc atcagcagtg aaagtgaaaa gagcagtcca
     4021 gaaaacatcc gatttaatag tgttgggtct cccatggaaa acaaccgaac aggacctgaa
     4081 agagtatttt agtacctttg gagaagttct tatggtgcag gtcaagaaag atcttaagac
     4141 tggtcattca aaggggtttg gctttgttcg ttttacggaa tatgaaacac aagtgaaagt
     4201 aatgtcacag cgacatatga tagatggacg atggtgtgac tgcaaacttc ctaattctaa
     4261 gcaaagccaa gatgagcctt tgagaagcag aaaagtgttt gtggggcgct gtacagagga
     4321 catgactgag gatgagctgc gggagttctt ctctcagtac ggggatgtga tggatgtctt
     4381 catccccaag ccattcaggg cctttgcctt tgttacattt gcagatgatc agattgcgca
     4441 gtctctttgt ggagaggact tgatcattaa aggaatcagc gttcatatat ccaatgccga
     4501 acctaagcac aatagcaata gacagttaga aagaagtgga agatttggtg gtaatccagg
     4561 tggctttggg aatcagggtg gatttggtaa tagcagaggg ggtggagctg gtttgggaaa
     4621 caatcaaggt agtaatatgg gtggtgggat gaactttggt gcgttcagca ttaatccagc
     4681 catgatggct gccgcccagg cagcactaca gagcagttgg ggtatgatgg gcatgttagc
     4741 cagccagcag aaccagtcag gcccatcggg taataaccaa aaccaaggca acatgcagag
     4801 ggagccaaac caggccttct ggtctggaaa taactcttat agtggctcta attctggtgc
     4861 agcaattggt gggggatcag catccaatgc agggtcgggc agtggtttta atggaggctt
     4921 tggctcaagc atggattcta agtcttctgg ctggggaatg tagacccagc tttcttgtac
     4981 aaagtggttg atcctctaga gctgagaact tcagggtgag tttggggacc cttgattgtt
     5041 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
     5101 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
     5161 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
     5221 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
     5281 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     5341 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     5401 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     5461 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     5521 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     5581 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     5641 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     5701 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     5761 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5821 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5881 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     5941 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     6001 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     6061 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     6121 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gactgcatta
     6181 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6241 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6301 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6361 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6421 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6481 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6541 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6601 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6661 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6721 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6781 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6841 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6901 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6961 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7021 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7081 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7141 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7201 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7261 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7321 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7381 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7441 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7501 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7561 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7621 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7681 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7741 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     7801 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7861 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7921 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7981 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8041 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8101 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8161 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8221 ctttcgtctt cactcgacgc ggccagcttg gtcgagctgg atacttcccg tccgccaggg
     8281 ggacatgccg gcgatgctga aggtcgcgcg cattcccgat gaagaggccg gttaccgcct
     8341 gttgacctgg tgggacgggc agggcgc
//
