The universal multiple cloning site
The universal multiple cloning site is designed to allow the versatile and easy cloning of gene fragments in one identical step with protein being directed into 5 different subcellular localizations with or without tags for identication and purification.

The multiple cloning site contains an NcoI site for clean in-frame cloning and BamHI as an alternative if NcoI is not available. Preceding the NcoI site there is an XbaI site which allows the use of the natural ATG of a cloned gene. For directional cloning making the best use of all the features of the vector family the available 3'-end sites are NotI or BglII followed by the vector-derived stopcodon with the options of a KDEL retention signal, and/or the identication and purification tag. Choosing this option will give you maximal flexibility and creates only three extra amino acids at the C-terminus of your protein (Asp-Leu-Gln). It requires that you subclone the gene in frame without its own stopcodon. Most proteins are not altered in their functional properties by the addition of such a short stretch of additional amino acids at the C-terminus. If only the N-terminal targeting options are of interest and tags are not desired than additional use can be made of the available SacI site.

A useful feature of the vector is the fact that BamHI and BglII (and BclI) produce compatible cohesive ends, so that cloning using these sites is completely interchangeable. Ready expression cassettes must be transferred to a plant expression vector such as pBINPLUS. Use can be made of the unique 8 cutters AscI and PacI, which occur very infrequently in genes.