LOCUS       sequence-432292-        5997 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPRcharm expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5997)
  AUTHORS   Neumann EN, Bertozzi TM, Wu E, Serack F, Harvey JW, Brauer PP, 
            Pirtle CP, Coffey A, Howard M, Kamath N, Lenz K, Guzman K, Raymond 
            MH, Khalil AS, Deverman BE, Minikel EV, Vallabh SM, Weissman JS
  TITLE     Brainwide silencing of prion protein by AAV-mediated delivery of an 
            engineered compact epigenetic editor.
  JOURNAL   Science. 2024 Jun 28;384(6703):ado7082. doi: 
            10.1126/science.ado7082. Epub 2024 Jun 28.
  PUBMED    38935715
REFERENCE   2  (bases 1 to 5997)
  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: "Science."; 
            date: "2024-06-28"; volume: "384(6703):ado7082. doi"; pages: " 
            10.1126/science.ado7082. Epub 2024 Jun 28"
COMMENT     Sequence Label: pENN226_CRISPRcharm
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1093..1113
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1114..1140,1141..1143,1144..3630,3631..3633,
                     3634..5217)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1114..5217
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5218..5238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5239..5295
                     /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"
ORIGIN
        1 atggccagga ccaaacaaac tgccagaaag agcaccgggg ggaaagcgcc ccgaaaacag
       61 ctggcgacca aagcagcccg caaaagcgca ccgggaggtg caagcagtgg ggcaggaagt
      121 tcctcaggag gaagtgcggc gggatccgga agcggtggcg catcaagtgg cgccgcaagt
      181 agcagtgccg ggtccgcagc tggctccggg tcctcccctg ttgagatcta caagaccgtg
      241 tccgcctgga agcggcagcc aatcagagtg ctgagcctgt tcggcaatat tgacaaggaa
      301 ctgaagtctc tgggcttcct ggaaagcggc agccacagcg agggcggcac actgaagtac
      361 gtggaagatg tgaccaacgt ggtgcggaga gatgtggaaa agtggggacc tttcgacctg
      421 ctgtacggca gcacccagcc tctgggcgga agctgcgaca ggtgtcctgg ctggtacatg
      481 ttccaatttc atagaatcct gcagtacgcc agacctagac aggagtctca gcccttcttc
      541 tggatcttca tggataacct gctgctcaca gaggacgacc agatgaccac cgtgcggttc
      601 ctgcaaacag aggccgtcac cctgcgcgac gtgagatcta gagtgctgca gaacgccgtc
      661 cgggtgtgga gcaacatccc cggtcttaaa agcaagcacg aggctctgtc cccaaaagag
      721 gaagagagcc tgcagggcca cgtgcggacc agatctaagc tggccgctcc taaagtggac
      781 cccctggtga agaactgcct gctgcctctg agagagtact ttaagtattt cagccagagc
      841 agcctgcccc tgggcggccc ttctagcggt gctccacctc caagcggcgg ctcgccagcc
      901 ggcagcccta ccagcaccga ggagggcaca agcgagagcg caacacctga gagcggcccc
      961 ggcacaagca ccgagccttc tgaaggaagc gcccctggga gccctgccgg cagccccacc
     1021 agcacagaag agggcacctc taccgagccc agcgagggta gcgcccctgg cacaagcaca
     1081 gaaccctctg agcctaagaa aaagcggaag gtgatggaca agaagtacag catcggcctg
     1141 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1201 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1261 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1321 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1381 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1441 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1501 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     1561 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     1621 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     1681 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     1741 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     1801 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     1861 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     1921 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     1981 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     2041 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     2101 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     2161 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2221 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2281 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     2341 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     2401 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2461 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     2521 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     2581 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     2641 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     2701 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     2761 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     2821 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     2881 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     2941 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     3001 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     3061 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     3121 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3181 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3241 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3301 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3361 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3421 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3481 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     3541 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     3601 atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac
     3661 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     3721 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     3781 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     3841 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     3901 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     3961 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     4021 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     4081 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     4141 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4201 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4261 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4321 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4381 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4441 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4501 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     4561 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     4621 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     4681 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     4741 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     4801 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     4861 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     4921 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     4981 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     5041 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     5101 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     5161 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggtgacccc
     5221 aagaagaagc ggaaggtggc cacaaatttc agcctgctga agcaggccgg cgacgtggaa
     5281 gagaatcctg gccccagcga gctgatcaaa gagaacatgc acatgaagct gtacatggaa
     5341 ggcaccgtgg acaaccatca cttcaagtgc accagcgagg gcgagggcaa gccctacgag
     5401 ggcacacaga ccatgagaat caaggtggtg gaaggaggcc cactgccctt cgccttcgac
     5461 atcctggcca ccagctttct gtacggctca aagaccttca tcaaccacac ccagggcatc
     5521 cctgatttct tcaagcagag cttccctgag ggatttacct gggagagagt gacaacttat
     5581 gaagatggcg gagtgctgac cgccacacaa gataccagcc tgcaggacgg ctgtctgatc
     5641 tacaacgtga agatcagagg cgtgaacttc accagcaacg gccctgtgat gcagaaaaaa
     5701 accctgggct gggaggcctt tacagagaca ctgtaccccg ctgatggagg cctggaaggt
     5761 agaaacgaca tggccctgaa actggtcggc ggcagccacc tgattgccaa tatcaagacc
     5821 acataccgga gcaagaaacc agctaaaaac ctgaagatgc ctggcgttta ctacgtggat
     5881 tacagactgg agcggatcaa ggaagccaac aacgagacat atgtggaaca gcacgaggtg
     5941 gccgtagcta gatactgcga cctgccttct aagcttggcc acaagctgaa ctgataa
//
