Difference between revisions of "Part:BBa K1150063"
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|'''RFC standard''' | |'''RFC standard''' | ||
| − | | | + | |[https://parts.igem.org/Help:Assembly_standard_25 RFC 25] |
|- | |- | ||
|'''Backbone''' | |'''Backbone''' | ||
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==Usage== | ==Usage== | ||
| − | + | At the front of this intermediate part you can ligate any favored promoter (Freiburg 2013 used SV40 and CMV promoters). At the back of this part you can fuse an effector domain without frameshift by using the RFC 25 standard. | |
<span class='h3bb'> | <span class='h3bb'> | ||
Revision as of 22:47, 4 October 2013
HA-NLS-Cas9-Linker 3 AA
.
| HA-NLS-Cas9-Linker | |
|---|---|
| Function | A new effector can be fused to
this Cas9 intermediate part |
| Use in | Mammalians |
| RFC standard | RFC 25 |
| Backbone | pSB1C3 |
| Submitted by | [http://2013.igem.org/Team:Freiburg Freiburg 2013] |
This intermediate part can be used to engineer your own dCas9 fusion proteins. [http://2013.igem.org/Team:Freiburg Freiburg 2013] already fused VP16, KRAB, G9a, UVR8, PIF6 and CRY2 to dCas9. With the HA tag you can verify via wester blot that your fusion protein is expressed or you can purify the protein with affinity chromatography. The NLS makes sure your fusion protein is transported into the nucleus of mammalian cells. The short 3 amino acids linker will make a connection beween dCas9 and your effector domain.
Usage
At the front of this intermediate part you can ligate any favored promoter (Freiburg 2013 used SV40 and CMV promoters). At the back of this part you can fuse an effector domain without frameshift by using the RFC 25 standard.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 312
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Literature