From 2007.igem.org

PCR MyoOF/BglIIR on Myoglobin ( 225bp=A) PCR BglIIF/XhoIR on Myoglobin (259bp=B) PCR XhoIF/MyoOR on Myoglobin ( 101bp=C)

PCR MyoOF/MyoOR on A+B+C (2566 bp,BglII/XhoI) Digest pBca9145-Bca1144 (BglII/XhoI,2054+910,L)

Places BglII site on edge of gene:Outside Forward cgttaAGATCTATGGTTCTGTCTGAAGGTGAATGG. Places XhoI site on edge of gene: Outside Reverse: ggtcatCTCGAGgttacGGATCCTCATCCCGAGCCACCCTGGTAACCC Mutates BglII site : BglII Forward: GAAAGCTTCTGAAGACCTGAAAAAACATGGTGTTAC Mutates BglII site: BglII Reverse: GTAACACCATGTTTTTTCAGGTCTTCAGAAGCTTTC Mutates XhoI site: XhoI Forward:GCTATGAACAAAGCTCTAGAGCTGTTCCGTAAAGA Mutates XhoI site:XhoI Reverse: TCTTTACGGAACAGCTCTAGAGCTTTGTTCATAGC