SBB09 Construct19

Construction of Cellulose-binding Knottin {} basic part M10046 (Bdvs003) PCR Odvs003F/Odvs003R on CBK                       (160bp, EcoRI/BamHI) Sub into pBca9495AK-Bca1144#5                      (EcoRI/BamHI, 3171+910, L) Product is pBca9495AK-M10046          {}       (3315bp)

Odvs003F   Forward construction of CBK basic part       ccataGAATTCATGAGATCTGGTTCTGGTCCGACCCAGTCTCACTACGG Odvs003R   Reverse construction of CBK basic part       ctgatGGATCCACCAGAACCCAGGCACTGAGAGTAGTACGG

JCA: There is no template for knottin, it needs to be synthesis. See revised below:

Combine 10pmol each of Odvs003,Odvs004,Odvs005,Odvs006 Run 55 program Use product as template for:

PCR Odvs003/Odvs006 on assembly product (160bp, EcoRI/BamHI) Product is pBca9495AK-M10046         {} -- Odvs003	Gene Synthesis of CBK basic part	CCATAGAATTCATGAGATCTGGTTCTGGTCCGACCCAGTCTCACTACGGTCAGTGCG Odvs004	Gene Synthesis of CBK basic part	CGCAAACGGTCGGACCAGAGTAACCGATACCACCGCACTGACCGTAGTGAGACTG Odvs005	Gene Synthesis of CBK basic part	CTCTGGTCCGACCGTTTGCGCTTCTGGTACCACCTGCCAGGTTCTGAACCCGTACTAC Odvs006	Gene Synthesis of CBK basic part	GCTTAGGATCCACCAGAACCCAGGCACTGAGAGTAGTACGGGTTCAGAACCTGGC