LOCUS       sequence-371894-        3469 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  vector for encoding and AAV packaging a human codon-optimized 
            High-fidelity RfxCas13d (hfCas13d) driven by EFS promoter, 
            Pcsk9-target guide RNAs compatible with RfxCas13d driven by hU6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3469)
  AUTHORS   Tong H, Huang J, Xiao Q, He B, Dong X, Liu Y, Yang X, Han D, Wang Z,
            Wang X, Ying W, Zhang R, Wei Y, Xu C, Zhou Y, Li Y, Cai M, Wang Q, 
            Xue M, Li G, Fang K, Zhang H, Yang H
  TITLE     High-fidelity Cas13 variants for targeted RNA degradation with 
            minimal collateral effects.
  JOURNAL   Nat Biotechnol. 2022 Aug 11. pii: 10.1038/s41587-022-01419-7. doi: 
            10.1038/s41587-022-01419-7.
  PUBMED    35953673
REFERENCE   2  (bases 1 to 3469)
  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: "Nat 
            Biotechnol. 2022 Aug 11. pii: 10.1038/s41587-022-01419-7. doi: 
            10.1038/s41587-022-01419-7."
COMMENT     Sequence Label: 
            AAV2-ITR-pEFS-NLS_hfCas13d(RfxCas13d_N2V8)_FLAG_NLS-pA-U6-DR-Pcsk9-s
            g5-DR-Pcsk9-sg6-DR-AAV2-ITR
FEATURES             Location/Qualifiers
     source          1..3469
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        44..255
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3200..3223
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3224..3244
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3245..3469
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
       61 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg
      121 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
      181 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
      241 gccgccagaa cacaggaccg gtgccaccat gcctaaaaag aaaagaaagg tgggttctgg
      301 tatcgagaag aagaagagct tcgccaaggg catgggagtg aagagcaccc tggtgtccgg
      361 ctctaaggtg tacatgacca catttgctga gggaagcgac gccaggctgg agaagatcgt
      421 ggagggcgat agcatcagat ccgtgaacga gggagaggct ttcagcgccg agatggctga
      481 caagaacgct ggctacaaga tcggaaacgc caagttttcc cacccaaagg gctacgccgt
      541 ggtggctaac aacccactgt acaccggacc agtgcagcag gacatgctgg gactgaagga
      601 gacactggag aagaggtact tcggcgagtc cgccgacgga aacgataaca tctgcatcca
      661 ggtcatccac aacatcctgg atatcgagaa gatcctggtg gagtacatca caaacgtggt
      721 gtacgtggtg aacaacatct ccggcctgga caaggatatc atcggcttcg gaaagttttc
      781 taccgtgtac acatacgacg agttcaagga tccagagcac caccgggccg cttttaacaa
      841 caacgacaag ctgatcaacg ccatcaaggc tcagtacgac gagttcgata actttctgga
      901 taaccccagg ctgggctact tcggacaggc tttcttttct aaggagggca gaaactacat
      961 catcaactac ggaaacgagt gttacgacat cctggccctg ctgagcggac tgaggcactg
     1021 ggtggtgcac aacaacgagg aggagtctcg gatcagccgc acctggctgt acaacctgga
     1081 caagaacctg gataacgagt acatctccac actgaactac ctgtacgaca ggatcaccaa
     1141 cgagctgaca aacagcttct ccaagaactc tgccgctaac gtgaactaca tcgctgagac
     1201 cctgggcatc aacccagctg agttcgctga gcagtacttc agattttcca tcatgaagga
     1261 gcagaagaac ctgggcttca acatcacaaa gctgagagaa gtgatgctgg acagaaagga
     1321 tatgtccgag atcaggaaga accacaaggt gttcgattct atcagaacca aggtgtacac
     1381 aatgatggac tttgtgatct acaggtacta catcgaggag gatgccaagg tggccgctgc
     1441 caacaagagc ctgcccgaca acgagaagtc tctgagcgag aaggatatct tcgtgatcaa
     1501 cctgagaggc tcctttaacg acgatcagaa ggacgctctg tactacgatg aggccaacag
     1561 gatctggaga aagctggaga acatcatgca caacatcaag gagttccggg gaaacaagac
     1621 ccgcgagtac aagaagaagg acgctccaag gctgcctagg atcctgcctg ctggaaggga
     1681 cgtgagcgcc ttcagcaagc tgatgtacgc cctgacaatg tttctggacg gaaaggagat
     1741 caacgatctg ctgaccacac tgatcaacaa gttcgacaac atccagtctt ttctgaaagt
     1801 gatgcctctg atcggcgtga acgctaagtt cgtggaggag tacgccttct ttaaggacag
     1861 cgccaagatc gctgatgagc tgcggctgat caagtccttt gccaggatgg gagagccaat
     1921 cgctgacgct aggagagcta tgtacatcga tgccatccgg atcctgggaa ccaacctgtc
     1981 ttacgacgag ctgaaggctc tggccgacac cttcagcctg gatgagaacg gcaacaagct
     2041 gaagaagggc aagcacggaa tgcgcaactt catcatcaac aacgtgatca gcaacaagcg
     2101 gtttcactac ctgatcagat acggcgaccc agctcacctg cacgagatcg ctaagaacga
     2161 ggccgtggtg aagttcgtgc tgggacggat cgccgatatc cagaagaagc agggccagaa
     2221 cggaaagaac cagatcgacc gctactacga gacctgcatc ggcaaggata agggaaagtc
     2281 cgtgtctgag aaggtggacg ctctgaccaa gatcatcaca ggcatgaact acgaccagtt
     2341 cgataagaag agatctgtga tcgaggacac cggaagggag aacgccgaga gagagaagtt
     2401 taagaagatc atcagcctgt acctgacagt gatctaccac atcctgaaga acatcgtgaa
     2461 catcaacgct agatacgtga tcggcttcca ctgcgtggag cgcgatgccc agctgtacaa
     2521 ggagaaggga tacgacatca acctgaagaa gctggaggag aagggcttta gctccgtgac
     2581 caagctgtgc gctggaatcg acgagacagc ccccgacaag aggaaggatg tggagaagga
     2641 gatggccgag agagctaagg agagcatcga ctccctggag tctgctaacc ctaagctgta
     2701 cgccaactac atcaagtact ccgatgagaa gaaggccgag gagttcacca ggcagatcaa
     2761 cagagagaag gccaagaccg ctctgaacgc ctacctgagg aacacaaagt ggaacgtgat
     2821 catccgggag gacctgctgc gcatcgataa caagacctgt acactgttcc ggaacaaggc
     2881 tgtgcacctg gaggtggctc gctacgtgca cgcctacatc aacgacatcg ccgaggtgaa
     2941 ctcctacttt cagctgtacc actacatcat gcagaggatc atcatgaacg agagatacga
     3001 gaagtctagc ggcaaggtgt ctgagtactt cgacgccgtg aacgatgaga agaagtacaa
     3061 cgatagactg ctgaagctgc tgtgcgtgcc tttcggatac tgtatcccac ggtttaagaa
     3121 cctgagcatc gaggccctgt tcgaccgcaa cgaggctgcc aagtttgata aggagaagaa
     3181 gaaggtgagc ggcaactccg attacaaaga cgatgacgat aagcccaaga agaagaggaa
     3241 agtcctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3301 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3361 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3421 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatgg
//
