Coding

Part:BBa_K2130008

Designed by: Ooi Kean Hean   Group: iGEM16_NTU-Singapore   (2016-10-19)
Revision as of 06:54, 19 October 2016 by OOIK0003 (Talk | contribs)

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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]


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Categories
Parameters
None