Difference between revisions of "Part:BBa K2130008"

Line 8: Line 8:
 
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'          NNNNNNNNNNNNNNNNAAAA 5'

Revision as of 06:53, 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]