We offer two basic vector constructs that allow the AviTag to be placed on either the C (pAC series) or N (pAN series) terminus of a desired protein, in E. coli. These third generation vectors have been engineered to substantially enhance biotinylation rates of the AviTag

 
 
 
 
 
 
 
 
 
 
N-terminus AviTag vectors

pAN Series (N terminus vectors)
pAN series of vectors are used to express N-terminal AviTag-protein fusions. The AviTag peptide sequence used is the #85 sequence described in Schatz (1993). In April of 1999, Avidity improved the N-terminal AviTag vectors to include a glutamic acid residue that has been shown to greatly enhance biotinylation rates of the AviTag. (Beckett et al., 1999)

Three versions of the pAN series vector exist: pAN-4 pAN-5 pAN-6 allowing any gene to be placed in frame with the AviTag. Expression of the AviTagged gene is under the control of the tightly repressed Trc promoter and is IPTG-inducible. (induction protocol). The vectors confer ampicillin resistance

Note: All genes inserted into the multiple cloning site MUST contain a stop codon to allow for desired protein. It is recommended that the start codon of the protein of interest be removed.

Below are the vector maps and sequences of each of the three pAN vectors in either Vector NTI format or text. Please use diagrams in planning your cloning strategies

pAN 4
pan4.txt
 
pAN 5
pan5.txt
 
pAN 6
pan6.txt
 
RE Cut Sites
RE pan.pdf

 

 



Printable file of map