Composite
His-FluA-S

Part:BBa_K243010:Design

Designed by: Freiburg Bioware09   Group: iGEM09_Freiburg_bioware   (2009-10-14)
Revision as of 17:04, 21 October 2009 by Timo (Talk | contribs) (→‎Design Notes)

His-FluA-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 cloning steps were planed theoretically before we started the work in the wet lab. The combiantion of HisTag linked with a FluA Tag connect by the Split Linker approved as a good way to express the inactive protein domain of our universal endonuclease.

Freiburg09 Hisfluasplfoki.jpg

part of universal restriction enzyme. Blue: DNA strand; Red: 16bp long Oligos, tag as indicated in the picture.
Ochre: Fluorescein A binding lipocalin;light Blue: inactive FokI cleavage domain;

We used the split linker, becaues it is an proved part of the team Freiburg08 and it is a suitable linker for fusion proteins. The properties of the split linker are an good compromise between for the stability and the distance for the connection between protein domain Fok_a and the anticalin tag. The parts are fused with RFC 25.
Commented GenBank file

Source

Combined the parts by serial cloning steps.

References