LOCUS       sequence-208223-        8422 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 8422)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 8422)
  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: pHBS1330 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 WT]
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(279..298)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     548..566
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2188..2197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        2350..2653
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2654..2857
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2808..2828
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2919..2928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2925..3626
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     misc_feature    3627..3692
                     /label=MCS
                     /note="multiple cloning site"
     CDS             3717..3773
                     /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(5019..5043)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          5071..5643
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(5775..5794)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5840..5895
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5894..5913)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6293..6310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6544..6563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7223..8014,8015..8083))
                     /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     7846..7865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8084..8188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8256..8274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttacttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg
       61 ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta
      121 ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta
      181 gtggccgccg tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc
      241 ggggtacatc cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc
      301 gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag
      361 ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca
      421 cttgaagccc tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac
      481 gtaggccttg gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg
      541 gccacccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctgagg
      601 aaaaaaaagg aaagggcatg ctcgccatgt gcaagataca gtgttgaatg tggtgcaaaa
      661 tataaagatg tggaagaaaa agccactgaa aataatatga ggaaataaaa aactagagta
      721 aattatcagt ggaacaattt ctattataca agacactaca cccatacatt ctcctaatgc
      781 tcatgtaaga atttctccaa aaataccttc cagcactaca aactagaaaa aaaaagagac
      841 atggacacca aattaagttc ttaatagtga agaacaaaag aactaccttg cctgctccag
      901 tggatccagc aaccgccaac aactgtcctc tttctatctt gaaattaata tctttcagga
      961 caggagtacc aagaagtgag aaattactga agaagaggct gtcatcacca ttagaagttt
     1021 ttctattgtt attgttttgt tttgctttct caaattattc cccaaatccc tgttaaaaaa
     1081 aaacacacac acacacacac acacacacat caaaaataaa agatgagttt gtcagatatt
     1141 ttcaatagat gcttgtttta tgtaccatac tcacctcgcc ctcgccctcg atctcgaact
     1201 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
     1261 ccatgttatc ctcctcgccc ttgctggttc cggtgcctcc actttcatct tcacctttag
     1321 tgataccggc tgccgtcacg aactccttca ggaccatgtg atcgcgcttc tcgttcgggt
     1381 ctttgctcag cgtggtctgg gtgctcagat agtggttgtc cggcaggcgc accgggccgt
     1441 cgccgatcgg ggtgttctgc tgtttgtggt cggcttcctg ctggctgccg tcctcaacgt
     1501 tgtggcggat cttgaagtta gccttgatgc cgttcttctg cttgtcggct tcgatacgga
     1561 cgttgtggct gttgtggttg tactccagct tgtgacccag gatgttgccg tcctccttga
     1621 agtcgttgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctccg
     1681 cgcgggtctt gtaggtgccg tcgtccttga actcgatggt gcgctcctgg acgtagcctt
     1741 ccggcatggc ggacttgaag aagtcgtgac gcttcatgtg gtccgggtag cggctgaagc
     1801 actgcacgcc gtaggtcagg gtggtcacga gggtcggcca cggcaccggc agcttgccgg
     1861 tggtgcagat gaacttcagg gtcagcttgc cctcggtggc atcgccctcg ccctcgccac
     1921 gcacgctgaa cttgtggccg tttacgtcgc cgtccagctc gactttgata ggcacagtac
     1981 ctgtaaatag ttcctctcct ttactggtac catcatcagg gaatgagctt acatttctcc
     2041 tcttcagcat ctggtcatcc ttcttagctt tcctcagctc cacattgacc tctattctgc
     2101 gacgcctcat ttctgtactg tcttttccct tgttcttgaa tctgtgaaga cgggcagctg
     2161 gtgtattagc attctcgttg gtggatccca tggtggcacc ggtgaattct ccaggcgatc
     2221 tgacggttca ctaaacgagc tctgcttata taggcctccc accgtacacg ccacctcgac
     2281 atactcgagt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2341 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2401 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2461 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2521 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2581 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2641 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     2701 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2761 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     2821 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
     2881 ctagggatcc tctagtcagc tgacgcgtta ccaccggtgc caccatggga agcgagctga
     2941 ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca
     3001 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
     3061 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg
     3121 gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc
     3181 ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
     3241 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
     3301 acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg
     3361 agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg
     3421 tgggcgggag ccatctgatc gcaaacgcca agaccacata tagatccaag aaacccgcta
     3481 agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg
     3541 ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc
     3601 ctagcaaact ggggcacaag cttaattccg gactcagatc tcgagctcaa gcttcgaatt
     3661 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagaggagga agcggagcta
     3721 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctatgtctg
     3781 aatatattcg ggtaaccgaa gatgagaacg atgagcccat tgaaatacca tcggaagacg
     3841 atgggacggt gctgctctcc acggttacag cccagtttcc aggggcgtgt gggcttcgct
     3901 acaggaatcc agtgtctcag tgtatgagag gtgtccggct ggtagaagga attctgcatg
     3961 ccccagatgc tggctgggga aatctggtgt atgttgtcaa ctatccaaaa gataacaaaa
     4021 gaaaaatgga tgagacagat gcttcatcag cagtgaaagt gaaaagagca gtccagaaaa
     4081 catccgattt aatagtgttg ggtctcccat ggaaaacaac cgaacaggac ctgaaagagt
     4141 attttagtac ctttggagaa gttcttatgg tgcaggtcaa gaaagatctt aagactggtc
     4201 attcaaaggg gtttggcttt gttcgtttta cggaatatga aacacaagtg aaagtaatgt
     4261 cacagcgaca tatgatagat ggacgatggt gtgactgcaa acttcctaat tctaagcaaa
     4321 gccaagatga gcctttgaga agcagaaaag tgtttgtggg gcgctgtaca gaggacatga
     4381 ctgaggatga gctgcgggag ttcttctctc agtacgggga tgtgatggat gtcttcatcc
     4441 ccaagccatt cagggccttt gcctttgtta catttgcaga tgatcagatt gcgcagtctc
     4501 tttgtggaga ggacttgatc attaaaggaa tcagcgttca tatatccaat gccgaaccta
     4561 agcacaatag caatagacag ttagaaagaa gtggaagatt tggtggtaat ccaggtggct
     4621 ttgggaatca gggtggattt ggtaatagca gagggggtgg agctggtttg ggaaacaatc
     4681 aaggtagtaa tatgggtggt gggatgaact ttggtgcgtt cagcattaat ccagccatga
     4741 tggctgccgc ccaggcagca ctacagagca gttggggtat gatgggcatg ttagccagcc
     4801 agcagaacca gtcaggccca tcgggtaata accaaaacca aggcaacatg cagagggagc
     4861 caaaccaggc cttcggttct ggaaataact cttatagtgg ctctaattct ggtgcagcaa
     4921 ttggttgggg atcagcatcc aatgcagggt cgggcagtgg ttttaatgga ggctttggct
     4981 caagcatgga ttctaagtct tctggctggg gaatgtagac ccagctttct tgtacaaagt
     5041 ggttgatcct ctagagctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc
     5101 tttttcgcta ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt
     5161 agaatgggaa gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac
     5221 tttctactct gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt
     5281 tcgttaaact ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca
     5341 gattgtaagt actttctcta atcacttttt tttcaaggca atcagggtat attatattgt
     5401 acttcagcac agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg
     5461 catataaatt ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc
     5521 atcatcctgc ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa
     5581 tactctgagt ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta
     5641 cagctcctgg gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcact
     5701 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     5761 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     5821 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     5881 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     5941 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     6001 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     6061 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     6121 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     6181 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgactg cattaatgaa
     6241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     7861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8281 gtcttcactc gacgcggcca gcttggtcga gctggatact tcccgtccgc cagggggaca
     8341 tgccggcgat gctgaaggtc gcgcgcattc ccgatgaaga ggccggttac cgcctgttga
     8401 cctggtggga cgggcagggc gc
//
