Device

Part:BBa_K1150047

Designed by: Natalie Louis and Lisa Schmunk   Group: iGEM13_Freiburg   (2013-09-17)
Revision as of 15:42, 24 September 2013 by NatalieL. (Talk | contribs)

Truncated CMV dCas9 Device #1

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. Truncation 1: here 365bp at the end 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]


[edit]
Categories
Parameters
None