LOCUS       sequence-203696-        9643 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 9643)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 9643)
  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: pHBS1203 [IBB-GFP-mCherry3E]-[BFP-P2A-TDP43 2x-Phi]
FEATURES             Location/Qualifiers
     source          1..9643
                     /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          6309..6881
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    7078..7133
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7524..7541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7695..8283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7775..7794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8454..9245,9246..9314))
                     /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     9077..9096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9315..9419)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9487..9505
                     /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 taaacataat agcaaccgac aactcgagag atctggtaga ggtaatccga
     5821 atcagaattc ccgaggagga tttgggggag ggtttggagg cgggttcggc cttgggatgg
     5881 gcttcggttt ttctggagcg ggcaacaatc agggctcaaa cggtggtatg aacgcaataa
     5941 acccggctat gatggctgcg gctcaggccg cactacagag cagttgggga atgatgggca
     6001 tgctggctag tcagcagaat cagagtggtc cgtcagggaa caaccagaac cagggaaata
     6061 tgcagcgcga gcctaatcag gctagcggaa acaattcatt tggttattca atcggatggg
     6121 ggggctttgg gggatttgga gggtggggca tgtcaggcag taatagcgga gccgcgagcg
     6181 cttccaatgc agggtcaggt tcgaattcta tggacagcaa aagttcgtag acccagcttt
     6241 cttgtacaaa gtggttgatc ctctactagt aagcttgcat gcgatatctc tagactgaga
     6301 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gaaaaattca
     6361 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     6421 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     6481 tgtctcctct tattttcttt tcattttctg taactttttt cgttaaactt tagcttgcat
     6541 ttgtaacgaa tttttaaatt cactttcgtt tatttgtcag attgtaagta ctttctctaa
     6601 tcactttttt ttcaaggcaa tcagggtaat tatattgtac ttcagcacag ttttagagaa
     6661 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     6721 gaaatattct tattggtaga aacaactaca tcctggtaat catcctgcct ttctctttat
     6781 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     6841 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     6901 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     6961 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     7021 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctgggtaat
     7081 aaaggaaatt tattttcatt gcaatagtgt gtgggaattt tttgtgtctc tcactcggaa
     7141 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     7201 caacatatgc catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat
     7261 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     7321 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     7381 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     7441 cttctcttat gaactcgact gcattaatga atcggccaac gcgcggggag aggcggtttg
     7501 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7561 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7861 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8101 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8401 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8461 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8521 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8581 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8641 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8701 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8761 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8821 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8881 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8941 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     9001 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     9061 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9301 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9421 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     9481 tataaaaata ggcgtatcac gaggcccttt cgtcttcact cgacgctggt cgagctgata
     9541 cttcccgtcc gccaggggac atgccggcga tgctgaaggt cgcgcgcatt cccgatgaag
     9601 aggccggtta ccgcctgttg acctggtggg acgggcaggg cgc
//
