Difference between revisions of "Part:BBa K1150055"
Line 4: Line 4:
 
Behind a strong SV40 promoter a HA-tag, a NLS and dCas9 followed by another NLS and a BGH terminator were assembled via the iGEM cloning method.  
 
Behind a strong SV40 promoter a HA-tag, a NLS and dCas9 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 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.  
 
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.  
−
 
  
  

Revision as of 10:37, 4 October 2013

Truncated SV40 dCas9 Device #4

Behind a strong SV40 promoter a HA-tag, a NLS and dCas9 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 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. Truncation 4: 1928pb in the middle 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 664
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]