Difference between revisions of "Part:BBa K2340000"
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<b><font size="+1"> Usage and Biology </font></b>
 
<b><font size="+1"> Usage and Biology </font></b>
  
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'dead' CAS13a with a double HEPN nuclease 1 and 2 inactive mutation, linked to a green fluorescent protein (wtGFP) via a linker sequence (ggctcctccggc). On both ends of the construct there are nuclear localization sequences (NLS; cccaagaaaaaacgcaaggtg) that will reduce background noise when expressed and used in mammalian cell lines.
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'dead' CAS13a with a double HEPN nuclease 1 and 2 inactive mutation, linked to a green fluorescent protein (wtGFP) via a linker sequence (ggctcctccggc). On the N-terminal and the C-terminal of the construct there are nuclear localization sequences (NLS; DNA sequence: cccaagaaaaaacgcaaggtg. Amino acid sequence: PKKKRKV) that will reduce background noise when expressed and used in mammalian cell lines.
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When supplied with a guide RNA (gRNA), this enzyme will process it into a CRISPR RNA (crRNA). The crRNA contains a sequence that is complementary to the RNA it is targetting.
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Revision as of 16:39, 28 October 2017


dCAS13a linked to GFP with NLS


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1112
    Illegal BglII site found at 2048
    Illegal BglII site found at 2312
    Illegal BglII site found at 2816
    Illegal BglII site found at 3302
    Illegal BglII site found at 3410
    Illegal BglII site found at 3740
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 662
    Illegal SapI.rc site found at 3421


Usage and Biology

'dead' CAS13a with a double HEPN nuclease 1 and 2 inactive mutation, linked to a green fluorescent protein (wtGFP) via a linker sequence (ggctcctccggc). On the N-terminal and the C-terminal of the construct there are nuclear localization sequences (NLS; DNA sequence: cccaagaaaaaacgcaaggtg. Amino acid sequence: PKKKRKV) that will reduce background noise when expressed and used in mammalian cell lines.

When supplied with a guide RNA (gRNA), this enzyme will process it into a CRISPR RNA (crRNA). The crRNA contains a sequence that is complementary to the RNA it is targetting.