Composite

Part:BBa_K1445001:Design

Designed by: Josephina Hendrix   Group: iGEM14_CU-Boulder   (2014-09-25)

Endogenous Type II CRISPR-Cas9 phagemid


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2179
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4458
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5400
    Illegal BsaI.rc site found at 5377


Design Notes

We have experienced some toxic effects by the pCas9 part so we suggest growing part-containing cells in lower concentrations of antibiotic, even if plasmid contains a high copy origin of replication.


Source

The M13 origin of replication from Litmus28i and part BBa_K1218011 from the registry.

References

Jiang, Wenyan, et al. 2013. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol.