Difference between revisions of "Part:BBa K243010:Design"
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DNA strand; Red: 16bp long Oligos, tag as indicated in the picture.<br> Ochre: | DNA strand; Red: 16bp long Oligos, tag as indicated in the picture.<br> Ochre: | ||
Fluorescein A binding lipocalin;light Blue: inactive FokI cleavage domain;</p> | Fluorescein A binding lipocalin;light Blue: inactive FokI cleavage domain;</p> | ||
| − | We used the split linker, becaue 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 | + | We used the [https://parts.igem.org/Part:BBa_K157009 split linker], becaue it is an proved part of the team [http://2008.igem.org/Team:Freiburg Freiburg08] and it is a suitable linker for fusion proteins. The properties of the split linker are an good compromise between the stability and the distance for the connection between protein domain Fok_a and the anticalin tag. |
The parts are fused with [https://parts.igem.org/Assembly_standard_25 RFC 25].<br> | The parts are fused with [https://parts.igem.org/Assembly_standard_25 RFC 25].<br> | ||
[https://static.igem.org/mediawiki/parts/6/6a/HisFluASplitFoki.txt Commented GenBank file] | [https://static.igem.org/mediawiki/parts/6/6a/HisFluASplitFoki.txt Commented GenBank file] | ||
Revision as of 17:10, 21 October 2009
His-FluA-Split Linker-Fok_i
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 272
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
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, becaue it is an proved part of the team [http://2008.igem.org/Team:Freiburg Freiburg08] and it is a suitable linker for fusion proteins. The properties of the split linker are an good compromise between 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.