Difference between revisions of "Part:BBa K243010:Design"
(→Design Notes) |
|||
| (One intermediate revision by the same user not shown) | |||
| Line 12: | Line 12: | ||
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 [https://parts.igem.org/Part:BBa_K157009 split linker] because it is an improved part of the team [http://2008.igem.org/Team:Freiburg Freiburg08] and a suitable linker for fusion proteins. The properties of the split linker are a good compromise between the stability and the distance for the connection between protein domain Fok_a and the anticaline tag. | + | We used the [https://parts.igem.org/Part:BBa_K157009 split linker] because it is an improved part of the team [http://2008.igem.org/Team:Freiburg Freiburg08] and a suitable linker for fusion proteins. The properties of the split linker are a good compromise between the stability and the distance for the connection between protein domain Fok_a and the anticaline tag.The dimerization partner to this part needs another combination of anticaline and the protein domain Fok_a, because the labeled oligonucleotides had different anticalins. |
The parts are fused according to [https://parts.igem.org/Assembly_standard_25 RFC 25].<br> | The parts are fused according to [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] | ||
Latest revision as of 03:09, 22 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 planned theoretically before we started the work in the wet lab.
The combination of HisTag linked with a FluA tag connected 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 because it is an improved part of the team [http://2008.igem.org/Team:Freiburg Freiburg08] and a suitable linker for fusion proteins. The properties of the split linker are a good compromise between the stability and the distance for the connection between protein domain Fok_a and the anticaline tag.The dimerization partner to this part needs another combination of anticaline and the protein domain Fok_a, because the labeled oligonucleotides had different anticalins.
The parts are fused according to RFC 25.
Commented GenBank file
Source
Combined the parts by serial cloning steps.