OtsBA-I716303

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PCR otsBF/otsBIR on otsBA (485 bp, gp = A) PCR otsBIF/otsAIR on otsBA (720 bp, gp = B) PCR otsAR/otsAIF on otsBA (413 bp, gp = C)

PCR otsBF/otsAR on A+B+C (2200 bp, BglII/XhoI) Digest Pbca9145-Bca1144 (BglII/XhoI, 2054+910, L) Product is Pbca9145- BBa I716303

otsBF Forward BglII site anneals to otsB cgtaaAGATCTATGACAGAACCGTTAACCGA otsBIF Removes internal BglII site GCAACGTATTACTCAGATtTGGCCACAAATGGCG otsBIR Removes internal BglII site CGCCATTTGTGGCCAaATCTGAGTAATACGTTGC otsAIF Removing BamHI site in otsA CGATGACATTATCTGGATtCACGATTATCACCTG otsAIR Removing BamHI site in otsA CAGGTGATAATCGTGaATCCAGATAATGTCATCG otsaR reverse XhoI site anneals to otsA cgtatCTCGAGttaGGATCCTTACGCAAGCTTTGGAAAGGTAGC

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