LOCUS       sequence-239565-        7843 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  MFASS reporter with wild-type exon ENSE00001732647.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7843)
  TITLE     Many rare genetic variants have unrecognized large-effect 
            disruptions to exon recognition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7843)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: RCP175: single exon control
FEATURES             Location/Qualifiers
     source          1..7843
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     580..602
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     664..683
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      727..736
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2703..2759
                     /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"
     polyA_signal    3524..3748
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3927..4742
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3927..4742
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4000..4019)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4913..5501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5402..5421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5655..5672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5789..5810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5825..5830,5831..5848,5849..5855)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     5874..5896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5893..5909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5893..5909
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5958..6079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5969..5988)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6023..6042
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6210..6809)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6293..6313)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6790..6809
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    6816..6864
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     enhancer        6904..7207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     gap             7349..7371
                     /estimated_length=23
ORIGIN
        1 gcggtgcccc gcggtgcggg ggggactgcg aggggaacaa aggctgcgtg cggggtgtgt
       61 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
      121 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
      181 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
      241 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
      301 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
      361 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
      421 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
      481 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
      541 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
      601 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
      661 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg atttgatacc
      721 gcgggcgcca ccatggtgtc taagggcgaa gagctcttta ctggcgtggt gcccatcctg
      781 gttgaattgg atggagacgt taatggacac aaatttagcg tctctggaga gggcgaagga
      841 gatgctactt atggcaaact gacactgaaa tttatttgca ctaccggcaa actgcccgta
      901 ccgtggccca cactggtgac aacatttaca tacggagtgc aggtactggc tggattgggt
      961 tagggaaacc gaggcggttc gctgaatcgg gtcgagcact tggcggagac gcgcgggcca
     1021 actacttagg gacagtcatg aggggtaggc ccgccggctg cagcccttgc ccatgcccgc
     1081 ggtgatcccc atgctgtgcc agcctttgcc cagaggcgct ctagctggga gcaaagtccg
     1141 gtcactgggc agcaccaccc cccggacttg catgggtagc cgctgagatg gagcctgagc
     1201 acactgttaa cgcagtgttt ctctaacttt caccggtttg atcacttggt taaggtggta
     1261 tctgtcaggt ttctccactg aaaagttact atttttccag caatactttg agactagcat
     1321 gctctaattt ttcataggtg gttgttctta tgaatattga tctgcttcgg cagcaacagc
     1381 gctggaaaga tggattacaa gaattgagaa ctggcttagc aactgtagaa gcacaggtag
     1441 agtatgtttt attttgtgtg tatactttga aatatgacct tttcagaatg taagtacaat
     1501 atatgaattg ccgagaagtt cataaaagtt atctttgcta gcgcaagagt tccagccggg
     1561 ctatggccag tgagatccaa gaatcttcct gtctctgctg atccactgat aggattacaa
     1621 gtacatgcca ccaagcccag cttcctctta ccaggtgctg gggaccaaac ttaggccctc
     1681 attcctacac agtgaatact tgactttgtt atcacccaac cctaataaat aactcactat
     1741 ccaaacaagt tgaaaccctt agaattctgt gttgctccag catgatgttg tggtaaacgt
     1801 taatacaata agatgcacag gtcataagtg cacattagct aagtgttgac aaagacttag
     1861 acctacataa cttaacccta ttagccctcc agaaagttcc tcattctcca ttccaggcaa
     1921 ctttcatcac accacatcat gtacaactac tattgaagtt gttttccact atagatacaa
     1981 tgagatgtca catacggctt tgtgttttga tttgcaagta ccaatcgagt atgaaatatg
     2041 gagtggatat tggacattgg ccaccatcta aatactttgt gttaaaagaa ttggttttca
     2101 taatttgttt tgtactgact gctggctagt cagattacct gactagtatg gacaggattt
     2161 tgcaataatc ataattcttt tttcagtgct tcgccagata tcccgatcac atgaaacagc
     2221 acgatttctt taagagcgcc atgcctgagg gttacgtgca ggagcgaaca attttcttca
     2281 aagacgatgg caattacaaa actcgagcag aagtgaagtt tgaaggggac actttagtta
     2341 acagaataga actgaagggg attgatttca aagaagacgg gaatattcta ggtcataagc
     2401 tcgaatacaa ctacaatagc cacaaagtct atatcaccgc tgataagcag aagaacggga
     2461 tcaaagtgaa cttcaaaaca cgccataaca ttgaggacgg gagcgtgcaa ctagccgacc
     2521 attaccaaca aaatacacca atcggagatg gccctgtgtt gttgcctgac aaccactatt
     2581 tgagcacgca gtccgcactg agcaaagatc cgaatgaaaa gagggaccac atggtcctgc
     2641 tggagttcgt tactgccgct ggcataacac ttggaatgga tgagctatat aaaggaagcg
     2701 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctg
     2761 tttccaaagg agaagaagat aacatggcga taatcaagga gttcatgaga ttcaaagtgc
     2821 acatggaagg aagcgttaac ggccatgagt tcgagattga aggcgaaggc gaagggcgac
     2881 catacgaggg gacacagaca gctaaattga aagtgaccaa aggaggaccc cttcccttcg
     2941 cttgggatat cctgagccca cagtttatgt atggaagcaa ggcatatgta aagcatcccg
     3001 cagacattcc agattacctt aagctctcct tccccgaggg attcaagtgg gagagagtca
     3061 tgaactttga ggatggcggc gtggttacag tcacacagga ctccagtctg caggatggag
     3121 aattcattta taaagtcaaa ctgaggggga ccaactttcc tagcgatggt ccagtgatgc
     3181 agaagaaaac aatgggctgg gaggcctctt cagagaggat gtatccagaa gatggggctc
     3241 ttaagggaga gatcaaacag agattaaagt tgaaagacgg tggtcattat gatgctgaag
     3301 tgaagactac atataaggcc aagaaaccag tccagctccc cggcgcatat aatgtcaaca
     3361 ttaaactcga cattacctcc cataacgaag attacaccat agtcgaacaa tatgagaggg
     3421 ctgagggcag gcacagtaca ggcgggatgg acgaattgta caagtaaggg cccgtttaaa
     3481 cccgctgatc agcctcgact gtgccttcta ggatccggcc cgactgtgcc ttctagttgc
     3541 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3601 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3661 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3721 catgctgggg atgcggtggg ctctatggcc agatatacgc gttgatagca gacactctat
     3781 gcctgtgtgg gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtatga gccatattca acgggaaacg tcttgctcta
     3961 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     4021 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     4081 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     4141 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     4201 atgatgcatg gttactcacc actgcgatcc ctgggaaaac agcattccag gtattagaag
     4261 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     4321 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     4381 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     4441 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     4501 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     4561 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     4621 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     4681 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     4741 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4801 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4861 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4921 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4981 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5041 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5101 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5161 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5221 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5281 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5341 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5401 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5461 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5521 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5581 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5641 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5701 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5761 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5821 aggctttata ctttatgctt ccggctcgta tgttgtgtgg aattgtgaag cttagcggat
     5881 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     5941 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     6001 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     6061 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     6121 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     6181 cggtggaatc accgggcttg gtagaattct caggcaccgg gcttgcgggt catgcaccag
     6241 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     6301 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
     6361 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     6421 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     6481 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6541 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6601 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6661 tgcagctcgg tgacccgctc gatgtggcgg tccgggtcga cggtgtggcg cgtggcgggg
     6721 tagtcggcga acgcggcggc gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg
     6781 aggcgcaccg tgggcttgta ctcggtcatg gtggcggccg gatgatcctg acgacggaga
     6841 ccgccgtcgt cgacaagccg gccggtaccg taggtagact atagggcgct ggcggccgct
     6901 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6961 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     7021 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7081 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     7141 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7201 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     7261 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     7321 ggggggcgcg cgccaggcgg ggcggggcnn nnnnnnnnnn nnnnnnnnnn naggcggaga
     7381 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     7441 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
     7501 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     7561 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
     7621 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
     7681 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
     7741 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     7801 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggg
//
