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EcoRI: 5' GAATTC 3' CTTAAG XbaI: 5' T*CTAGA 3' AGATC*T SpeI: 5' A*CTAGT 3' TGATC*A PstI: 5' CTGCAG 3' GACGTC
First primer Extended Start codon Stop codon Restriction site Mutagenesis primer 1-FFT: