![](https://parts.igem.org/images/partbypart/icon_primer.png)
Primer
Part:BBa_K4615000:Design
Designed by: Xinyi (Marry) Xuan Group: iGEM23_Toronto (2023-10-12)
Forward primer to insert 20 bp of sgRNA of tpiA into pTargetF
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 27
Illegal XbaI site found at 33
Illegal PstI site found at 45 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 27
Illegal PstI site found at 45 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 27
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 27
Illegal XbaI site found at 33
Illegal PstI site found at 45 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 27
Illegal XbaI site found at 33
Illegal PstI site found at 45 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We obtained this part by introducing 20 bp of sgRNA within the tpiA genome that guides the Cas9 protein to cut and create strand breaks.
Source
The 20bp of sgRNA was generated using CHOPCHOP and analyzed based on various factors such as GC content, self-complementarity and efficiency.