Extremely Efficient, Accurate and Flexible vector to clone your genes and express your Proteins of Interest
pOnebyOne™ are extremely efficient, accurate and flexible Bicistronic Mammalian Expression Kits that contains an expression cassette based in 2A sequence Breakthrough technology.
pOnebyOne™ FREE allows co-expression of two genes of interest driven by Human Cytomegalovirus early promoter. It is based in 2A sequence from Thosea asigna Virus (TaV) which is flanked by two multiple cloning sites. T2A sequence leads to the highest level of Protein Expression at the second gene position compared to IRES sequence.
The mechanism of 2A self-cleavage is a ribosome skipping process where the ribosome skips the formation of a glycil-prolyl peptide bond at the C-terminus of 2A. Stable mammalian cells could be selected by neomycin resistance.
Both genes of interest must be in frame with T2A and between them. Successful skipping results in translation of two proteins: 2A upstream protein is attached to the 2A sequence (GSGEGRGSLLTCGDVEENPG) at C-terminus while 2A downstream protein is attached to one Pro at the N-terminus.