LOCUS       sequence-371897-        3322 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  vector for encoding a human codon-optimized dead Cas13X (dCas13X) 
            driven by CBh promoter, guide RNAs compatible with Cas13X driven by 
            hU6, EGFP driven by SV40 promoter, and mCherry driven by SV40 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3322)
  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 3322)
  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: 
            pCBh_NLS_dCas13X_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCher
            ry-pA
FEATURES             Location/Qualifiers
     source          1..3322
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        288..565
                     /label=chicken beta-actin promoter
     intron          566..794
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             815..835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3158..3178
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3188..3322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3225..3244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3279..3298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      121 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      181 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
      241 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      301 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      361 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
      421 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      481 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      541 taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg ctgccttcgc cccgtgcccc
      601 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      661 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt
      721 ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa
      781 tcactttttt tcaggttgga ccggtgccac catgcccaag aagaagcgga aggtggccca
      841 ggtgagcaag cagacctcca agaagaggga gctgagcatc gacgagtacc agggcgcccg
      901 gaagtggtgc ttcaccattg ccttcaacaa ggccctggtg aaccgggaca agaacgacgg
      961 cctgttcgtg gaaagcctgc tgagacacga gaagtacagc aagcacgact ggtacgacga
     1021 agatacccgg gccctgatca agtgcagcac ccaggccgcc aacgccaagg ctgaagccct
     1081 ggcgaactac ttcagtgctt accggcatag ccctggctgc ctgaccttca ccgccgagga
     1141 cgaactgcgg accatcatgg agagagccta tgagcgggcc atcttcgagt gcagaagaag
     1201 agagacagag gtgatcatcg agtttcccag cctgttcgag ggcgaccgga tcaccaccgc
     1261 cggcgtggtg tttttcgtga gctttttcgt ggaaagaaga gtgctggatc ggctgtatgg
     1321 agccgtgtcc ggcctgaaga agaatgaggg acagtacaag ctgacccgga aggccctgag
     1381 catgtactgc ctgaaggaca gcagattcac caaggcctgg gataagcggg tgctgctgtt
     1441 cagagacatc ctggcccagc tgggaagaat ccccgccgag gcctacgagt actaccacgg
     1501 cgagcagggt gataagaaga gagctaacga caatgagggc acaaatccca agcggcacaa
     1561 ggacaagttc atcgaatttg cactgcacta cctggaagcc cagcacagcg agatctgctt
     1621 cggcagacgc cacatcgtgc gggaagaggc cggcgccggc gatgagcaca agaagcaccg
     1681 gaccaaggga aaggtggtgg tggacttcag caagaaggac gaggaccaga gctactatat
     1741 ctccaagaac aacgtgatcg tgcggatcga caagaacgcc ggccctagaa gctaccggat
     1801 gggcctgaac gagctgaagt acctcgtgct gctgagcctg caggggaagg gcgacgatgc
     1861 catcgccaag ctgtacagat acagacagca cgtggagaac atcctggatg tggtgaaggt
     1921 gaccgataag gataaccacg tgttcctgcc ccgcttcgtg ctggagcagc acggcatcgg
     1981 cagaaaggcc ttcaagcagc ggatcgatgg acgggtgaag cacgtgcggg gcgtgtggga
     2041 gaagaagaag gccgccacca atgaaatgac cctgcacgag aaggccagag acatcctgca
     2101 gtacgtgaac gaaaactgca cccggtcctt caaccctggc gaatacaaca gactgctggt
     2161 gtgcctggtg ggcaaggacg tggagaactt tcaggccggc ctgaagcggc tgcagctggc
     2221 cgaaaggatc gatggccggg tgtactccat cttcgcccag accagcacca tcaatgagat
     2281 gcaccaggtg gtgtgcgacc agatcctgaa ccggctgtgc agaatcggcg accagaagct
     2341 gtacgattac gtgggactgg gcaagaagga cgaaatcgac tacaagcaga aggtggcctg
     2401 gttcaaggag cacatcagca tccggagagg attcctgaga aagaagttct ggtacgatag
     2461 caagaaggga ttcgcaaagc tggtggagga acacctggag tccggcggcg gccagcgcga
     2521 cgtgggcctg gacaagaagt actaccacat cgacgccatc ggcagattcg agggcgccaa
     2581 ccccgccctg tacgagaccc tggccagaga tcggctgtgc ctcatgatgg cccagtactt
     2641 cctgggcagc gtgagaaagg aactgggcaa caagattgtg tggagcaacg acagcatcga
     2701 actgcctgtg gaaggctctg tgggaaatga gaagagcatc gtgttctccg tgtctgacta
     2761 cggcaagctg tacgtgctgg acgatgccga attcctgggc cggatctgcg aatacttcat
     2821 gccccacgaa aagggcaaga tccggtacca cacagtgtac gaaaagggct ttagagcata
     2881 caacgacctg cagaagaagt gcgtggaggc cgtgctggct ttcgaagaga aggtggtgaa
     2941 ggccaagaag atgagcgaga aggaaggcgc ccactacatc gacttccggg agatcctggc
     3001 ccagaccatg tgcaaggagg ccgaaaagac cgcagtgaac aaggtggcgg ctgccttctt
     3061 cgctgcggcc ctgaagttcg tgattgacga gttcggcctg ttcagcgacg tgatgaagaa
     3121 gtacggcatc gagaaggaat ggaagttccc tgtcaagccc aagaagaagc ggaaggtgta
     3181 gaagcttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3301 tgtatcttat catgtctgga tc
//
