Difference between revisions of "Part:BBa K1150048"

Revision as of 09:11, 25 September 2013

Truncated CMV dCas9 Device #2

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 Cas9 gene was performed by PCR over the backbone (=Cas9 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.

Usage and Biology

This part can be used in the uniCAS Toolkit of team Freiburg as a shorter version of Cas9.

Truncation2: here 306bp of the anterior part of Cas9 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 576
Illegal BglII site found at 900
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K1150048 short</partinfo>
-.
+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 Cas9 gene was performed by PCR over the backbone (=Cas9 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.
-<!-- 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 Cas9.
+Truncation2: here 306bp of the anterior part of Cas9 are missing
 <!-- -->