Description
The three HiBiT CMV-neo Flexi® Vectors are configured to facilitate simple, efficient transfer of the gene of interest into a vector designed for genetic attachment of the HiBiT peptide tag to the amino or carboxy terminus of the protein of interest using the Flexi® Cloning System ( Cat.# C8640 ). The vectors can be used for both stable and transient gene expression and encode kanamycin resistance for bacterial selection and neomycin resistance for mammalian selection.
The pFC37K HiBiT CMV-neo Flexi® Vector appends the HiBiT to the C terminus of the gene of interest.
The pFN38K HiBiT CMV-neo Flexi® Vector adds the HiBiT tag to the N terminus of the gene of interest.
The pFN39K secHiBiT CMV-neo Flexi® Vector attaches the HiBiT tag to the N terminus of the gene of interest. This vector encodes the IL-6 secretion signal peptide N-terminal to the HiBiT tag for direct trafficking of HiBiT-tagged proteins to the plasma membrane of mammalian cells for cell surface expression and secreted proteins. Note: We recommend removing naturally encoded secretion signal sequences from the gene of interest for efficient cell-surface expression of the HiBiT-tagged protein.
The HiBiT peptide tag, in combination with the Nano-Glo® HiBiT detection systems, offers bioluminescent detection of the protein of interest. This results in quantitation of proteins using bioluminescence with no need for antibodies. Notes:
Expression of the HiBiT-tagged protein will only occur when the proper reading frame is maintained between the HiBiT tag and the gene of interest.
The HiBiT peptide sequence is provided for reference only. To obtain rights to synthesize the HiBiT tag, please see the Terms and Conditions of Use .
About the Flexi® Vector System
The Flexi® Vector System is a simple, yet powerful, 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. Directly use recombinant clones and minimize time wasted screening background colonies. The versatile cloning of the Flexi® system means you can choose from a variety of expression systems and fusion tag orientations and then transfer to others as required. Direct transfers can only occur between two N-terminal tagged vectors or from an N-terminal to a C-terminal vector.
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.
Ready-to-Use HaloTag® Flexi® Vector Clones
For access to 20,000 human and 15,000 mouse ORF clones with HaloTag® Technology in multiple mammalian expression vectors and custom cloning options, visit: www.kasuza.or.jp.kop/dsearch-e
pFC37K HiBiT CMV-neo Flexi® Vector sequence text file .
pFN38K HiBiT CMV-neo Flexi® Vector sequence text file .
pFN39K secHiBiT CMV-neo Flexi® Vector sequence text file .
