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Part:BBa_K5423002

Designed by: Kyle Cavanagh   Group: iGEM24_ANU   (2024-10-02)


mScarlet3 dCas9 gRNA

This part codes for a mScarlet3 targetting gRNA insert with BsaI sites. The gRNA insert was designed with chop chop to target mScarlet3. The BsaI sites allow it to be inserted into the dCas9 system. Despite confirmation of successful gRNA primer insertion, via enzyme digest, and sanger sequencing, we were unable to replicate the levels of repression observed in the source paper1, with initial testing showing less than 20% percent reduction. Following this, we attempted to optimize for [anhydrotetracycline] and [L-arabinose], but this was also unsuccessful, with changing concentrations resulting in decreased activity relative to the source paper1conditions

highara-hist-final.png rha-atc-altered-hist-final.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5
    Illegal BsaI.rc site found at 41


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Categories
Parameters
None