SBB09 Construct5

Construction of {a~Int-native>} basic part PCR Bhs001F/Bhs002R on E. coli strain 0157:H7    (145 bp, gp = A) PCR Bhs002F/Bhs003R on E. coli strain 0157:H7     (1888 bp, gp = B)

PCR Bhs001F/Bhs003R on A+B                      (2008 bp, EcoRI/BamHI) Digest pBca9495CA-Bca1144#5                     (EcoRI/BamHI, 3039+910, L) Product is pBca9495CA-M10024                     {a~Int-native>}

Bhs001F Forward EcoRI for {a~Int-native>}           cccaaGAATTCatgAGATCTtaacATGATTACTCATGGTTG Bhs002F Removing the EcoRI site from {a~Int-native>}   GTTAATCAGAAcTCATTTGCAAATG Bhs002R Removing the EcoRI site from {a~Int-native>}   CATTTGCAAATGAgTTCTGATTAAC Bhs003R Reverse BamHI for {a~Int-native>}           GCAAAggatccGCCTTGGTTTGATCAAAAAATATAACCGCAC

JCA: There is a frame error on Bhs003R. Revised: Construction of {a~Int-native>} basic part PCR Bhs001F/Bhs002R on E. coli strain 0157:H7    (146 bp, gp = A) PCR Bhs002F/Bhs003R on E. coli strain 0157:H7     (1889 bp, gp = B)

PCR Bhs001F/Bhs003R on A+B                      (2009 bp, EcoRI/BamHI) Digest pBca9495CA-Bca1144#5                     (EcoRI/BamHI, 3039+910, L) Product is pBca9495CA-M10024                     {a~Int-native>}

Bhs001F Forward EcoRI for {a~Int-native>}          cccaaGAATTCatgAGATCTtaacATGATTACTCATGGTTG Bhs002F Removing the EcoRI site from {a~Int-native>}   GTTAATCAGAACTCATTTGCAAATGG Bhs002R Removing the EcoRI site from {a~Int-native>}  CCATTTGCAAATGAGTTCTGATTAAC Bhs003R Reverse BamHI for {a~Int-native>}          GCAAAggatccGGCCTTGGTTTGATCAAAAAATATAACCGCAC