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 EcoRI site found at 7
Illegal EcoRI site found at 868 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 7
Illegal EcoRI site found at 868 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 7
Illegal EcoRI site found at 868 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 7
Illegal EcoRI site found at 868 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 7
Illegal EcoRI site found at 868 - 1000COMPATIBLE WITH RFC[1000]
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