Part:BBa_K3738021
Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6XHistidine Tag
Cas13a is an enzyme originating from Leptotrichia buccalis (Lbu) which functions to cleave single-stranded RNAs (ssRNAs); particularly mRNAs. This function is achieved following protein-RNA complex formation with CRISPR RNA (crRNA) via crRNA backbone contacts with residues from the Helical-2, HEPN1, and Linker domains of Cas13a. The crRNA contains a spacer region coding for a direct repeat stem loop as well as a region complementary to target ssRNAs. Once the enzyme complex interacts with a target ssRNA, a structural conformation change occurs within the domains of the protein that permits active site formation for non-discriminate ssRNA cleavage (O’Connel et al., 2019).
O'Connell MR. Molecular Mechanisms of RNA Targeting by Cas13-containing Type VI CRISPR–Cas Systems. Journal of Molecular Biology. 2019;431(1):66-87. doi: https://doi.org/10.1016/j.jmb.2018.06.029.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 398
Illegal BglII site found at 1334
Illegal BglII site found at 1544
Illegal BglII site found at 1598
Illegal BglII site found at 1829
Illegal BglII site found at 2102
Illegal BglII site found at 2588 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911
Illegal NgoMIV site found at 2050
Illegal NgoMIV site found at 2686 - 1000COMPATIBLE WITH RFC[1000]
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