Difference between revisions of "Part:BBa K1150061"

Latest revision as of 10:43, 4 October 2013

Truncated dCas9 VP16 Device #5

Behind a strong SV40 promoter a HA-tag, a NLS and dCas9 followed by another NLS, VP16 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 nucleotides 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.

Usage and Biology

This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of dCas9. with this device it can be testest if the truncated dCasß version can still activate genes via VP16. Truncation 5: 945bp at the end of dCas9 are missing

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 664
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K1150061 short</partinfo>
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+Behind a strong SV40 promoter a HA-tag, a NLS and dCas9 followed by another NLS, VP16 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 nucleotides 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.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of dCas9. with this device it can be testest if the truncated dCasß version can still activate genes via VP16.
+Truncation 5: 945bp at the end of dCas9 are missing
 <!-- -->