Plasmid pBS7

Plasmid pBS7 is built from pDH6 except YFP has been mutated to the Venus variant. Those mutations are described in "A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications" by Nagai, Ibata, Park, Kubota, Mikoshiba and Miyawaki in Nat Biotechnol 2002 Jan;20(1):87-90. In addition we have included Q69M, a mutation briefly mentioned in the Nagai et al. paper as potentially beneficial, although not necessary for the enhanced maturation.

Forward and reverse primers refer to the sequence of the primers described on the protocol page "Fusing GFP to the Carboxy Terminus of Your Favorite Protein". Some common restriction enzyme sites are shown. Unique sites are in red.

Map:

Mutations: F46L, F64L, S65G, V68L, Q69M, S72A, Q80R, M153T, V163A, S175G, T203Y
Q80R is believed to be a neutral mutation.

The DNA sequence of pBS7 is shown below with the Venus sequence in green.

Back to Plasmids & Protocols