DNA
stgRNA 1

Part:BBa_K4630000:Design

Designed by: Zhejun Qin   Group: iGEM23_WHU-China   (2023-10-06)


self-targeting guide RNA 1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1642
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3921
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 6510
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4863
    Illegal BsaI.rc site found at 4840


Design Notes

Based on a previous study, we adopted a one-plasmid system which we called pCas (pRed-Cas9-poxb300) that combines Cas9 and Lambda-Red recombinases in E. coli, which reported to achieve 100% genome editing efficiency in 6 hours of induction.1 We constructed this new composite part from the plasmid pCas , which we obtained from Genescript. We employed Golden Gate Assembly and Gibson Assembly to remove the gRNA sequence and the homologous arm sequence.


Source

Cas9 comes from Streptococcus pyogenes. Lambda-Red comes from Lambda phage.

References