Description
The three pBiT3.1 HiBiT Vectors are configured to append the 11 amino acid HiBiT peptide tag to the amino or carboxy terminus of the target protein. These vector contain a multiple cloning region to generate an in-frame HiBiT fusion protein. The pBiT3.1 Vectors can be used for both stable and transient gene expression and encode kanamycin resistance for bacterial selection and blasticidin resistance for mammalian selection. The flexible linker between the protein of interest and the HiBiT tag will vary in length, depending on the restriction enzyme used.
The pBiT3.1-N [CMV/HiBiT/Blast] Vector appends the HiBiT tag to the N terminus of the gene of interest. The insert should contain a stop codon at the 3´ end for termination of the translation.
The pBiT3.1-C [CMV/HiBiT/Blast] Vector adds the HiBiT tag to the C terminus of the gene of interest. Note: The insert should not encode a stop codon, and the gene of interest should contain proper translation initiation sequences, including an N-terminal ATG codon or Kozak sequence.
The pBiT3.1-secN [CMV/HiBiT/Blast] Vector attaches the HiBiT tag to the N terminus of the mature form of transmembrane or secreted proteins. 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 or secretion. Note: The insert should also contain a stop codon at the 3´ end for termination of the translation.
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 .
pBiT3.1-N [CMV/HiBiT/Blast] Vector sequence text file .
pBiT3.1-C [CMV/HiBiT/Blast] Vector sequence text file .
pBiT3.1-secN [CMV/HiBiT/Blast] Vector sequence text file .
