SBB09 Construct5
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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)
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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>}
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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
