Part:BBa_K243015:Design
Strep-DigA-Split Linker-Fok_i
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 278
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We applied the Streptavidine tag to enable a simultaneous purification of constructs with a His tag. Strep tag also shows a higher affinity towards Strep-Tactin than His tag. For that the purification with Strep-tag is more specific. The used DigA tag allows the coupling to an Digoxigenin linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligo and the construct containing the protein domain Fok_i is not as efficient as the use of a combination of FluA tagged oligo with a construct containing Fok_i. To avoid interactions between the DigA tag with the connected protein domain Fok_i we applied the Split Linker. This linker is an improved part of the team Freiburg08 and a suitable linker for fusion proteins. Commented GenBank file
Source
Combined the parts by serial cloning steps.