Figure 4. Restriction map of molecular construct sgRNA/Cas9/pTREX-n derived from vector Cas9/pTREX-n by insertion of a specific sgRNA sequence through BamHI restriction site.

Rib. Prom, ribosomal promoter; HX1 and gapdh are trans-splicing regions previously described (Vazquez and Levin, 1999); AmpR, ampicillin resistance gene; Neo, neomycin resistance gene; Cas9-HA-2xNLS-GFP is a fusion gene encoding for Cas9 endonuclease as previously described ( Lander et al., 2015 ).