resources
expression vectors
plasmids are available from addgene and MolecularCloud
These vectors are optimized for gene insertion by Gibson assembly, and all proteins will be purified using immobilized-metal affinity chromatography.
vectors for standard protein expression
pCDB736: N-terminal 10-His SUMO AviTag 5xGGS TEV-peptide fusion
the AviTag enables enzymatic ligation of biotin to the protein
scarless removal of the 10-His SUMO tag is accomplished using SUMO protease, leaving the AviTag attached to the protein N-terminus by a long, flexible linker
scarless removal of the entire fusion domain is accomplished using TEV protease
pCDB24 or pCDB179: N-terminal 10-His SUMO fusion
scarless removal of the His tag is accomplished using SUMO protease
a SUMO fusion can promote protein folding and solubility
pCDB347: N-terminal 10-His TEV-peptide fusion
scarless removal of the His tag is accomplished using TEV protease
pCDB97: N-terminal 6-His fusion
this His tag is not cleavable
pCDB99 or pCDB348: C-terminal 6-His fusion
this His tag is not cleavable
pCDB256: N-terminal 10-His fusion
this His tag is not cleavable
pCDB98: C-terminal 10-His fusion
this His tag is not cleavable
vectors for expression of proteins with one or more disulfide bonds
pCDB26 or pCDB180: N-terminal OsmY (for secretion) 10-His SUMO fusion
scar-less removal of the His tag is accomplished using SUMO protease
pCDB28 or pCDB380: N-terminal OsmY (for secretion) 10-His TEV-peptide fusion
pCDB364: N-terminal 10-His DsbC TEV-peptide fusion
this is a fusion to the mature form (i.e. the secretion signal peptide has been removed) of E. coli disulfide bond isomerase, which promotes expression and folding of proteins containing multiple disulfide bonds from the cytoplasm
scarless removal of the fusion domain is accomplished using TEV protease
pCDB424: N-terminal 10-His DsbC SUMO fusion
this is a fusion to the mature form of E. coli disulfide bond isomerase
scarless removal of the fusion domain is accomplished using SUMO protease
pCDB435: N-terminal 10-His DsbC TEV-peptide AviTag fusion
the AviTag enables enzymatic ligation of biotin to the protein, and this will remain attached to the inserted gene following digestion with protease and removal of the fusion domain
pCDB441: N-terminal 10-His DsbC SUMO AviTag fusion
miscellaneous expression tools