Coding
Part:BBa_K2019003:Design
Designed by: John Lazar Group: iGEM16_Northwestern (2016-10-12)
Periplasm Directed saCas9 via TorA Signaling Sequence (TorA-saCas9)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 753
Illegal BglII site found at 906
Illegal BglII site found at 1052 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 326
Illegal AgeI site found at 2882 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Part BBa_K1012002 was ordered as a gblock with homology to the BBa_K2019000-BBa_R0040 expression system. BBa_K2019000-BBa_R0040 was linearized at the 5' end of saCas9. A 2 piece gibson assembly was then performed with the gblock and Cas9 expression backbone.
Source
The TorA sequence is from part BBa_K1012002 of the 2013 Dundee Team. The saCas9 sequence is from another Northwestern 2016 iGEM part, BBa_K2019000.