LOCUS       sequence-203693-        9184 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 9184)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 9184)
  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: pHBS1108 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 ∆RRM]
FEATURES             Location/Qualifiers
     source          1..9184
                     /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(5772..5796)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          5850..6422
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6619..6674
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7065..7082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7236..7824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7316..7335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7995..8786,8787..8855))
                     /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     8618..8637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8856..8960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9028..9046
                     /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 aacatcctcc aatgccgaac ctaagcacaa tagcaataga cagttagaaa
     5341 gaagtggaag atttggtggt aatccaggtg gctttgggaa tcagggtgga tttggtaata
     5401 gcagaggggg tggagctggt ttgggaaaca atcaaggtag taatatgggt ggtgggatga
     5461 actttggtgc gttcagcatt aatccagcca tgatggctgc cgcccaggca gcactacaga
     5521 gcagttgggg tatgatgggc atgttagcca gccagcagaa ccagtcaggc ccatcgggta
     5581 ataaccaaaa ccaaggcaac atgcagaggg agccaaacca ggccttcggt tctggaaata
     5641 actcttatag tggctctaat tctggtgcag caattggttg gggatcagca tccaatgcag
     5701 ggtcgggcag tggttttaat ggaggctttg gctcaagcat ggattctaag tcttctggct
     5761 ggggaatgta gacccagctt tcttgtacaa agtggttgat cctctactag taagcttgca
     5821 tgcgatatct ctagactgag aacttcaggg tgagtttggg gacccttgat tgttctttct
     5881 ttttcgctat tgaaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta
     5941 gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact
     6001 ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt
     6061 tcgttaaact ttagcttgca tttgtaacga atttttaaat tcactttcgt ttatttgtca
     6121 gattgtaagt actttctcta atcacttttt tttcaaggca atcagggtaa ttatattgta
     6181 cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc
     6241 atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac atcctggtaa
     6301 tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat
     6361 actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc tttttcctac
     6421 agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc
     6481 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     6541 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     6601 agcatctgac ttctgggtaa taaaggaaat ttattttcat tgcaatagtg tgtgggaatt
     6661 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     6721 agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg
     6781 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     6841 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     6901 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     6961 ctcccagtca tagctgtccc tcttctctta tgaactcgac tgcattaatg aatcggccaa
     7021 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     7441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7861 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7921 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7981 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8041 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8101 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8161 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8221 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8281 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8341 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8401 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8461 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8521 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8581 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8641 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8701 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8761 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8821 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8881 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8941 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     9001 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
     9061 tcgacgctgg tcgagctgat acttcccgtc cgccagggga catgccggcg atgctgaagg
     9121 tcgcgcgcat tcccgatgaa gaggccggtt accgcctgtt gacctggtgg gacgggcagg
     9181 gcgc
//
