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Fig. 4 Schematics illustrating the construction of U6 sgRNA expression vector. All U6 vectors have a U6 promoter and a sgRNA scaffold. Digestion of BsmBI makes U6 vectors expose the overhangs recognizing the annealed sgRNA primer. Digestion of PstI and SalI can disrupt the vector undigested by BsmBI to improve positive cloning.
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