SBB09 Construct17

Construction of {} Basic Part M10022 (Bdvs001) into pBca9495AK PCR Odvs001F/Odvs002rR on VH             (379bp, gp = A) PCR Odvs002rF/Odvs001R on VH              ( 89bp, gp = B) --- PCR Odvs001F/Odvs001R on A+B to make Bdvs001       (436bp, EcoRI/BamHI) Sub M10022 into pBca9495KA-Bca1144#5               (EcoRI/BamHI, 3171+910, L) Product is pBca9495AK-M10022   {}           (3491bp) --- Odvs0007F  Forward EcoRI/BglII for Bglbrick of ECOHLY   ccataGAATTCATGAGATCTtattccgtggaagaacttattgggacc Odvs0007R  Reverse BamHI for Bglbrick of ECOHLY         ctgatGGATCCataaatatcattaccatatccacc Odvs0008rF Removing the BglII site in ECOHLY (Forward)  ggcagtgagggagcagaCctgcttgatggcgg Odvs0008rR Removing the BglII site in ECOHLY (Reverse)  ccgccatcaagcagGtctgctccctcactgcc

JCA: revised 1
Construction of beta roll intermediate PCR Odvs0007F/Odvs0008rR on CFT073 gen.        (379bp, gp = A) PCR Odvs0008rF/Odvs0007R on CFT073 gen.         ( 89bp, gp = B) --- PCR Odvs0007F/Odvs0007R on A+B to make Bdvs001       (436bp, EcoRI/BamHI) TA kit, product is pCR-betaroll --- Odvs0007F  Forward EcoRI/BglII for Bglbrick of ECOHLY   ccataGAATTCATGAGATCTtattccgtggaagaacttattgggacc Odvs0007R  Reverse BamHI for Bglbrick of ECOHLY         ctgatGGATCCataaatatcattaccatatccacc Odvs0008rF Removing the BglII site in ECOHLY (Forward)  ggcagtgagggagcagaCctgcttgatggcgg Odvs0008rR Removing the BglII site in ECOHLY (Reverse)  ccgccatcaagcagGtctgctccctcactgcc

Construction of {} Basic Part M10022 (Bdvs001) into pBca9495AK PCR Odvs0007F/ca1383 on CFT073 gen.        (304bp, gp = A) PCR ca1382/Odvs0007R on CFT073 gen.         (152bp, gp = B) --- PCR Odvs0007F/Odvs0007R on A+B to make Bdvs001       (436bp, EcoRI/BamHI) Sub M10022 into pBca9495KA-Bca1144#5               (EcoRI/BamHI, 3171+910, L) Product is pBca9495AK-M10022   {}           (3491bp) --- Odvs0007F  Forward EcoRI/BglII for Bglbrick of ECOHLY   ccataGAATTCATGAGATCTtattccgtggaagaacttattgggacc Odvs0007R  Reverse BamHI for Bglbrick of ECOHLY         ctgatGGATCCataaatatcattaccatatccacc ca1382	Forward oligo to remove ecoRI site in beta roll	GTTCAGGGcAATTCTCTTGC ca1383	Reverse oligo to remove ecoRI site in beta roll	GCAAGAGAATTgCCCTGAAC

JCA: revised 2
PCR Odvs0007F/ca1383 on CFT073 gen.          (304bp, gp = A) PCR ca1382/Odvs0008rR  on CFT073 gen.         (96bp, gp = B) PCR Odvs0008rF/Odvs0007R on CFT073 gen.       (89 bp, gp = C) --- PCR Odvs0007F/Odvs0007R on A+B+C          (436bp, EcoRI/BamHI) sub into pBca9495????? ..... --- Odvs0007F  Forward EcoRI/BglII for Bglbrick of ECOHLY   ccataGAATTCATGAGATCTtattccgtggaagaacttattgggacc Odvs0007R  Reverse BamHI for Bglbrick of ECOHLY         ctgatGGATCCataaatatcattaccatatccacc Odvs0008rF Removing the BglII site in ECOHLY (Forward)  ggcagtgagggagcagaCctgcttgatggcgg Odvs0008rR Removing the BglII site in ECOHLY (Reverse)  ccgccatcaagcagGtctgctccctcactgcc ca1382	Forward oligo to remove ecoRI site in beta roll	GTTCAGGGcAATTCTCTTGC ca1383	Reverse oligo to remove ecoRI site in beta roll	GCAAGAGAATTgCCCTGAAC