SBB09 Construct17
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Construction of {<ECOHLY>} Basic Part M10022 (Bdvs001) into pBca9495AK
PCR Odvs001F/Odvs002rR on VH (379bp, gp = A)
PCR Odvs002rF/Odvs001R on VH ( 89bp, gp = B)
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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 {<ECOHLY>} (3491bp)
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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 {<ECOHLY>} Basic Part M10022 (Bdvs001) into pBca9495AK
PCR Odvs0007F/ca1383 on CFT073 gen. (304bp, gp = A)
PCR ca1382/Odvs0007R on CFT073 gen. (152bp, gp = B)
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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 {<Beta Roll>} (3491bp)
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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
