Difference between revisions of "Part:BBa K3645016"
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Template of sgRNA. Use Golden Gate (BsaⅠ) to replace the template sequence with a ~20bp guide sequence. Add 'tagc' on the 5'-terminal of your guide sequence and 'aaac' on the reverse 5'-terminal. | Template of sgRNA. Use Golden Gate (BsaⅠ) to replace the template sequence with a ~20bp guide sequence. Add 'tagc' on the 5'-terminal of your guide sequence and 'aaac' on the reverse 5'-terminal. | ||
| − | 5'-tagc-20bp-****-3' | + | 5'-tagc-20bp-****-3'<br> |
3'-****-20bp-caaa-5' | 3'-****-20bp-caaa-5' | ||
Latest revision as of 07:17, 27 October 2020
sgRNA scaffold
Template of sgRNA. Use Golden Gate (BsaⅠ) to replace the template sequence with a ~20bp guide sequence. Add 'tagc' on the 5'-terminal of your guide sequence and 'aaac' on the reverse 5'-terminal.
5'-tagc-20bp-****-3'
3'-****-20bp-caaa-5'
Transcription product is a single strain sgRNA for dCas9.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 135
Illegal BsaI.rc site found at 37