Stop codon at the last codon of Cas9 followed with SV40-NLS - Cloning Specialist.

[Anan Tarabeih]

Greetings everyone!

I recently modified Cas9 and aim to incorporate it into another plasmid for nucleus expression, necessitating Cas9 to be followed by SV40-NLS. However, the presence of stop codons at the end of both Cas9 and NLS is impeding Cas9 expression and NLS signal inclusion (see attached screenshot). While I can potentially modify the stop codon, I'm uncertain if this could compromise the activity of Cas9.

Any insights on how to clone and express Cas9 effectively in the nucleus would be highly appreciated.

Best,

Anan Tarabeih