Part:BBa_K3791003

Spacer gRNA Kanamycin

This part contains the spacer sequence used to target a specific DNA fragment of a certain kanamycin-resistant gene. It must be used in combination with the proper repeat in order to conform the whole gRNA functionality [1]. More specifically, we placed it in a plasmid after the repeat sequence, already cataloged in the Parts registry. In our case, we used it for activating LbCas12a protein, so the repeat sequence appropriate for our design was BBa_K2927006. The whole gRNA construct containing both spacer and repeat is available in BBa_K3791008 and the same one but with the required promoter and terminator is available in BBa_K3791013.

Usage, Biology and Characterization

As this part is a simplification of our final construct, usage, biology and characterization is explained on both the preliminary gRNA construct part BBa_K3791013 and the Efficient gRNA construct part BBa_K3791023

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
  COMPATIBLE WITH RFC[1000]