Difference between revisions of "Part:BBa K2130008"
 
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Design Top strand of your spacer with 5' AGAT overhang, bottom strand with 5'AAAA overhang.
 
Design Top strand of your spacer with 5' AGAT overhang, bottom strand with 5'AAAA overhang.
  
−
Example:
+
Example:<br>
 
5' AGATNNNNNNNNNNNNNNNN          3'<br>
 
5' AGATNNNNNNNNNNNNNNNN          3'<br>
−
3'         NNNNNNNNNNNNNNNNAAAA 5'
+
3'&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;NNNNNNNNNNNNNNNNAAAA 5'
  
 
You can sequence you gRNA insertion with M13(-20)-R
 
You can sequence you gRNA insertion with M13(-20)-R

Latest revision as of 06:54, 19 October 2016


LbCpf1 gRNA expression cassette

The gRNA is expressed by the hU6 promoter. To insert gRNA, digest with BsmBI.

To ligate in gRNA: Design Top strand of your spacer with 5' AGAT overhang, bottom strand with 5'AAAA overhang.

Example:
5' AGATNNNNNNNNNNNNNNNN 3'
3'           NNNNNNNNNNNNNNNNAAAA 5'

You can sequence you gRNA insertion with M13(-20)-R



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 326
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]