Coding

Part:BBa_K3738021

Designed by: Emily Hagens and Mark Lea   Group: iGEM21_Lethbridge   (2021-10-19)
Revision as of 03:08, 20 October 2021 by Marklea (Talk | contribs)


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None