Composite

Part:BBa_K243018:Design

Designed by: Freiburg Bioware09   Group: iGEM09_Freiburg_bioware   (2009-10-14)

His-DigA-Split Linker-Fok_i


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 272
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The choice of the linker, lipocalin tag and purification tag allows us several different combinations. We applied the His tag to enable a simultaneous purification of constructs with a Strep tag. The used DigoxigeninA tag allows the coupling to an fluorescein linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligos and the construct containing the protein domain Fok_i is not that efficient as the use of a combination of FluA tagged oligos 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. The linker itself has no influence of the connected parts. We decided to use the Split Linker for this construct to get a longer distance between DigA tag and Fok_i to guarantee the independent function of both parts. This linker is an improved part of the team Freiburg08 and it is a suitable linker for fusion proteins. Commented GenBank file

Source

Combined the parts by serial cloning steps.

References