LOCUS       sequence-203702-        9617 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 9617)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 9617)
  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: pHBS1285 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 G3...
FEATURES             Location/Qualifiers
     source          1..9617
                     /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(6231..6255)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          6283..6855
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    7052..7107
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7498..7515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7669..8257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7749..7768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8428..9219,9220..9288))
                     /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     9051..9070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9289..9393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9461..9479
                     /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 taagcacaat agcaatagac agttagaaag aagtggaaga tttggtggta
     5821 atccaggtgg ctttgggaat cagggtggat ttggtaatag cagagggggt ggagctggtt
     5881 tgggaaacaa tcaaggtagt aatatgggtt ttgggatgaa ctttggtgcg ttcagcatta
     5941 atccagccat gatggctgcc gcccaggcag cactacagag cagttggggt atgatgggca
     6001 tgttagccag ccagcagaac cagtcaggcc catcgggtaa taaccaaaac caaggcaaca
     6061 tgcagaggga gccaaaccag gccttcggtt ctggaaataa ctcttatagt ggctctaatt
     6121 ctggtgcagc aattggttgg ggatcagcat ccaatgcagg gtcgggcagt ggttttaatg
     6181 gaggctttgg ctcaagcatg gattctaagt cttctggctg gggaatgtag acccagcttt
     6241 cttgtacaaa gtggttgatc ctctagagct gagaacttca gggtgagttt ggggaccctt
     6301 gattgttctt tctttttcgc tattgaaaaa ttcatgttat atggaggggg caaagttttc
     6361 agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc atgataattt
     6421 tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt cttttcattt
     6481 tctgtaactt ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
     6541 cgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
     6601 taattatatt gtacttcagc acagttttag agaacaattg ttataattaa atgataaggt
     6661 agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg tagaaacaac
     6721 tacatcctgg taatcatcct gcctttctct ttatggttac aatgatatac actgtttgag
     6781 atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc
     6841 ttctttttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc
     6901 aaagaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     6961 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     7021 atgaagcccc ttgagcatct gacttctggg taataaagga aatttatttt cattgcaata
     7081 gtgtgtggga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     7141 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca
     7201 tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     7261 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     7321 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     7381 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaactc gactgcatta
     7441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7861 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8101 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8461 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8521 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8581 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8641 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8701 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8761 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8821 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8881 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8941 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9001 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9061 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9121 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9181 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9241 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9301 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9361 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9421 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9481 ctttcgtctt cactcgacgc tggtcgagct gatacttccc gtccgccagg ggacatgccg
     9541 gcgatgctga aggtcgcgcg cattcccgat gaagaggccg gttaccgcct gttgacctgg
     9601 tgggacgggc agggcgc
//
