Coding
Part:BBa_K3738021:Design
Designed by: Emily Hagens and Mark Lea Group: iGEM21_Lethbridge (2021-10-19)
Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6XHistidine Tag
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 398
Illegal BglII site found at 1334
Illegal BglII site found at 1544
Illegal BglII site found at 1598
Illegal BglII site found at 1829
Illegal BglII site found at 2102
Illegal BglII site found at 2588 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911
Illegal NgoMIV site found at 2050
Illegal NgoMIV site found at 2686 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The anionic tag is present to facilitate uptake into our delivery particle MS2 whereas the Histidine tag was added for the purpose of Nickel-Affinity chromatography in obtaining the purified protein.
Source
The Cas13a protein comes from the Gram-negative bacteria Leptotrichia buccalis (Lbu).