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Fig. 1 Generation of pS/MAR-USH2A vectors (A) pS/MAR-USH2A vectors were generated by inserting the full USH2A coding sequence (16 kb) into the backbone in 5 fragments. The vectors contained a CAG or CMV promoter and P. plumata GFP sequence 5′ of the USH2A sequence and an S/MAR sequence at the 3′ end. The P2A sequence encodes a self-cleaving peptide. (B) PCR was performed to amplify each USH2A fragment. (C) pS/MAR backbone digestion cut out the puromycin cassette and restriction enzyme digestion controls of the pS/MAR-USH2A vectors using SAPI.
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Full text @ Mol. Ther.