This set of protein tags and destination vectors has been constructed to specifically work together, so that all elements always stay in frame. There are other tags in the collection, that were created for other combinations, that might work here as well, but always check first in silico if everything stays in frame! That will save you a lot of work!

Entry Vectors
A-B B-C C-D D-E E-F
pGG-A-GST-B pGG-B-GST-C pGG-C-GST-D pGG-D-GST-E pGG-E-GST-F
pGG-A-3xMYC-B pGG-B-3xMYC-C pGG-C-3xMYC-D pGG-D-3xMYC-E pGG-E-3xMYC-F
pGG-A-6xHIS-B pGG-B-6xHIS-C pGG-C-6xHIS-D pGG-D-6xHIS-E pGG-E-6xHIS-F
pGG-A-3xFLAG-B pGG-B-3xFLAG-C pGG-C-3xFLAG-D pGG-D-3xFLAG-E pGG-E-3xFLAG-F
pGG-A-3xHA-B pGG-B-3xHA-C pGG-C-3xHA-D pGG-D-3xHA-E pGG-E-3xHA-F
pGG-A-MBP-B pGG-B-MBP-C pGG-C-MBP-D pGG-D-MBP-E pGG-E-MBP-F

 

Entry Vectors - Others
Name Remarks
pGG-B-6xHIS-TEV-C Contains a TEV cleavage site
pGG-B-MBP-TEV-C Contains a TEV cleavage site
pGG-B-TEV-C Contains in-frame TEV protease cleavage site
pGG-D-TEV-E Contains in-frame TEV protease cleavage site
pGG-E-STOP-RBS-sfGFP-F The sequence of this fluorophore is preceded by a Stop codon and a ribosome binding site (RBS). This enables the construct to report translation from the same RNA of the construct of interest, serving as a proxy for expression.
pGG-E-STOP-RBS-mCherry-F The sequence of this fluorophore is preceded by a Stop codon and a ribosome binding site (RBS). This enables the construct to report translation from the same RNA of the construct of interest, serving as a proxy for expression.

 

Destination Vectors
Name Remarks
pDEST-RC-AG Golden gate adapted recombinant protein plasmid
pDEST-RC-BG Golden gate adapted recombinant protein plasmid
pDEST-RC-CG Golden gate adapted recombinant protein plasmid
pPICZ-AG This vector allows methanol inducible expression of the gene of interest in the cytoplasm of Pichia pastoris
pPICZα-AG This vector allows methanol inducible expression and protein secretion of the gene of interest using Pichia pastoris

 

Iterative Cloning
Name Remarks
pGG-F-T7-pT7-RBS-A-SacB-linker-G-G To be used in combination with pDEST-RC-AG, pDEST-RC-BG or pDEST-RC-CG
pGG-F-T7-pT7-RBS-B-SacB-linker-G-G To be used in combination with pDEST-RC-AG, pDEST-RC-BG or pDEST-RC-CG
pGG-F-T7-pT7-RBS-C-SacB-linker-G-G To be used in combination with pDEST-RC-AG, pDEST-RC-BG or pDEST-RC-CG