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Generation of Different Donor Templates and px459-mRFP Target
(A) One kilobase of double-stranded DNA (dsDNA) donor templates was generated with varying homology sequence overlaps on the 5′ and 3′ side of the mutation site. (B) Donor templates were generated as plasmid, linearized plasmid with 5′ or 3′ backbone overhang, or PCR product. (C) mRFP gene sequence was cloned into the px459 expression vector instead of the puromycin resistance gene.
