Gateway destination vector. Contains a synthetic core promoter followed by sequence encoding a human codon-optimized Cas9 RNA-guided nuclease that is tagged with nuclear localization signals at both the N-terminal and C-terminal ends of the open reading frame. A tissue-specific enhancer/promoter region can be introduced into this vector to generate a tissue-specific Cas9 expression construct which can then be inserted into the genome using phiC31:int integrase.