Part:BBa_K3738020
Lbu-Cas13a with an N-Terminal 6xHistidine Tag and C-Terminal Anionic 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 1089
Illegal PstI site found at 1944
Illegal PstI site found at 2883 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1089
Illegal PstI site found at 1944
Illegal PstI site found at 2883 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 370
Illegal BglII site found at 1306
Illegal BglII site found at 1516
Illegal BglII site found at 1570
Illegal BglII site found at 1801
Illegal BglII site found at 2074
Illegal BglII site found at 2560 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1089
Illegal PstI site found at 1944
Illegal PstI site found at 2883 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1089
Illegal PstI site found at 1944
Illegal PstI site found at 2883
Illegal NgoMIV site found at 2022
Illegal NgoMIV site found at 2658 - 1000COMPATIBLE WITH RFC[1000]
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