Coding

Part:BBa_K3799000

Designed by: Shubhamay Das   Group: iGEM21_IISER_Kolkata   (2021-09-30)


EiCsm6,A CRISPR system endoribonuclease

Description - EiCsm6 is obtained from Enterococcus italicus, and belongs to the type III CRISPR systems. Csm6 is mainly activated by cyclic oligoadenylate second messengers by binding to their CARF domain. This activates the HEPN domain of the Csm6 for collateral RNA degradation. The CARF domain then degrades the cyclic adenylate activator (cA6) which functions as an off switch to regulate the activity of the CRISPR enzyme. These mechanisms facilitate rapid invader clearance and ensures proper termination to limit self toxicity and suicide of the bacteria.

EiCsm6 is coupled with the enzyme LwaCas13a( BBa_K2323004) to create the detector system SHERLOCKv2. The overhangs that are produced by the cleavage of RNA by Cas13a produces 2’-3’ cyclic phosphate hex adenylate residues that activate Csm6. This activation results in the collateral cleavage of RNA by both Cas13a and Csm6. 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 314
    Illegal XhoI site found at 367
    Illegal XhoI site found at 1054
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//awards/basic_part
//cds
//function/crispr
Parameters
chassisE. Coli