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