Difference between revisions of "Part:BBa K1150050"
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===Usage and Biology=== | ===Usage and Biology=== | ||
This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of Cas9. | This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of Cas9. | ||
| − | Truncation 4: | + | Truncation 4: 1928pb in the middle of Cas9 are missing |
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Revision as of 19:38, 3 October 2013
Truncated CMV dCas9 Device #4 Behind a strong CMV promoter a HA-tag, a NLS and Cas9 in a shortened version followed by another NLS and a BGH terminator were assembled via the iGEM cloning method. The truncation of the huge dCas9 gene was performed by PCR over the backbone (=dCas9 in pSB1C3). The part between the primer pair is not amplified. One of the primers has an overlap compatible to the first bases of the other one, so that after this PCR a one fragment Gibson assembly could be performed.
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Usage and Biology
This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of Cas9. Truncation 4: 1928pb in the middle of Cas9 are missing
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 576
Illegal BglII site found at 900 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Functional Parameters
To verify the expression of the truncated dCas9 version we performed westernblot with anti-HA antibody.
