Composite
Part:BBa_K3738026:Design
Designed by: Emily Hagens and Mark Lea Group: iGEM21_Lethbridge (2021-10-21)
Lbu-Cas13a with an N-terminal 6xHistidine Tag and C-Terminal Anionic Tag and crRNA
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1134
Illegal PstI site found at 1989
Illegal PstI site found at 2928 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1134
Illegal PstI site found at 1989
Illegal PstI site found at 2928 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 415
Illegal BglII site found at 1351
Illegal BglII site found at 1561
Illegal BglII site found at 1615
Illegal BglII site found at 1846
Illegal BglII site found at 2119
Illegal BglII site found at 2605 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1134
Illegal PstI site found at 1989
Illegal PstI site found at 2928 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1134
Illegal PstI site found at 1989
Illegal PstI site found at 2928
Illegal NgoMIV site found at 2067
Illegal NgoMIV site found at 2703 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part has been codon optimized for E.coli. The part must be in vitro transcribed to obtain the direct-repeat stem loop RNA structure and complex with Cas13a.
Source
The Mcy gene cluster comes from Microcystic Aeruginosa, however the crRNA originates from Leptrotrichia buccalis. The Cas13a protein comes from the Gram-negative bacteria Leptotrichia buccalis (Lbu).