LOCUS       sequence-203720-        9340 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 9340)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 9340)
  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: pHBS1138 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43  ∆...
FEATURES             Location/Qualifiers
     source          1..9340
                     /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(5928..5952)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          6006..6578
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    6775..6830
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7221..7238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7392..7980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7472..7491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8151..8942,8943..9011))
                     /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     8774..8793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9012..9116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9184..9202
                     /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 gacctatgac atccgattta atagtgttgg gtctcccatg gaaaacaacc gaacaggacc
     5041 tgaaagagta ttttagtacc tttggagaag ttcttatggt gcaggtcaag aaagatctta
     5101 agactggtca ttcaaagggg tttggctttg ttcgttttac ggaatatgaa acacaagtga
     5161 aagtaatgtc acagcgacat atgatagatg gacgatggtg tgactgcaaa cttcctaatt
     5221 ctaagcaaag ccaagatgag cctttgagaa gcagaaaagt gtttgtgggg cgctgtacag
     5281 aggacatgac tgaggatgag ctgcgggagt tcttctctca gtacggggat gtgatggatg
     5341 tcttcatccc caagccattc agggcctttg cctttgttac atttgcagat gatcagattg
     5401 cgcagtctct ttgtggagag gacttgatca ttaaaggaat cagcgttcat atatccaatg
     5461 ccgaacctaa gcacaatagc aatagacagt tagaaagaag tggaagattt ggtggtaatc
     5521 caggtggctt tgggaatcag ggtggatttg gtaatagcag agggggtgga gctggtttgg
     5581 gaaacaatca aggtagtaat atgggtggtg ggatgaactt tggtgcgttc agcattaatc
     5641 cagccatgat ggctgccgcc caggcagcac tacagagcag ttggggtatg atgggcatgt
     5701 tagccagcca gcagaaccag tcaggcccat cgggtaataa ccaaaaccaa ggcaacatgc
     5761 agagggagcc aaaccaggcc ttcggttctg gaaataactc ttatagtggc tctaattctg
     5821 gtgcagcaat tggttgggga tcagcatcca atgcagggtc gggcagtggt tttaatggag
     5881 gctttggctc aagcatggat tctaagtctt ctggctgggg aatgtagacc cagctttctt
     5941 gtacaaagtg gttgatcctc tactagtaag cttgcatgcg atatctctag actgagaact
     6001 tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgaa aaattcatgt
     6061 tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc
     6121 accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt
     6181 ctcctcttat tttcttttca ttttctgtaa cttttttcgt taaactttag cttgcatttg
     6241 taacgaattt ttaaattcac tttcgtttat ttgtcagatt gtaagtactt tctctaatca
     6301 cttttttttc aaggcaatca gggtaattat attgtacttc agcacagttt tagagaacaa
     6361 ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa
     6421 atattcttat tggtagaaac aactacatcc tggtaatcat cctgcctttc tctttatggt
     6481 tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct
     6541 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggttg
     6601 ttgtgctgtc tcatcatttt ggcaaagaat tcactcctca ggtgcaggct gcctatcaga
     6661 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     6721 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct gggtaataaa
     6781 ggaaatttat tttcattgca atagtgtgtg ggaatttttt gtgtctctca ctcggaagga
     6841 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     6901 catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca tcagtatatg
     6961 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     7021 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     7081 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     7141 ctcttatgaa ctcgactgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7201 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7561 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7801 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8281 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9121 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     9181 aaaaataggc gtatcacgag gccctttcgt cttcactcga cgctggtcga gctgatactt
     9241 cccgtccgcc aggggacatg ccggcgatgc tgaaggtcgc gcgcattccc gatgaagagg
     9301 ccggttaccg cctgttgacc tggtgggacg ggcagggcgc
//
