cDNA cloning,protein expression,vectors,cloning,ORF,flexivector,g826,g832,g833,g838,6his, polyhistidine,dualtag
These vectors are used for inducible expression of polyhistidine-HaloTag fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. Use the Flexi Cloning System vectors for C- or N-terminal fusions.
The following vectors are used for inducible expression of HaloTag fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. Expression levels depend highly on the nature of the protein, but in general the N-terminal HaloTag fusion protein can increase expression level, enhance refolding and boost solubility of the expressed protein. HaloTag vectors are supplied as Flexi System vectors. The Flexi Vector System is a simple, directional cloning method for protein-coding sequences. It is based on two rare-cutting restriction enzymes, SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi Vectors without the need to resequence. Direct transfers can only occur between two N-terminal tagged vectors or from an N-terminal to a C-terminal vector. The Flexi system vectors contain a His6-HaloTag dual tag. The dual tag enables protein purification with the reusable Ni-resin while retaining the HaloTag covalent labeling properties. pFN29A/K His6HaloTag T7 Flexi Vectors (Cat.# G8261, G8331) are designed for protein expression with an N-terminal His6-HaloTag dual tag in E. coli T7 cell-free expression systems. pFC30A/K His6HaloTag T7 Flexi Vectors (Cat.# G8321, G8381) are designed for protein expression with a C-terminal His6-HaloTag dual tag in E. coli T7 cell-free expression systems. Note: Flexi Vectors contain the lethal barnase gene to reduce background colonies without inserts during the subcloning procedure. Using the Flexi Vector Cloning System replaces the barnase gene with your insert. These vectors, as purchased, cannot be cultured in normal laboratory strains of E. coli without an insert.