Table 7.

Various epitope tagged functional constructs of GIV

Construct	Expression vector (mammalian/bacterial)	Tag/position	Important observations	Cited References
Full-length GIV	p3XFLAG-CMV-14 and pcDNA3.1 expression vectors for transient or stable intracellular C-terminal 3XFLAG or Myc tag, respectively.	C-terminal FLAG and Myc tags	N-terminal tags mislocalize protein in cells, alter protein functions. Bulky C-terminal tags (GFP etc.), even with the placement of flexible linkers, express poorly.	GIV-FLAG in p3XFLAG-CMV-14 (ADDGENE ID# 65947) (Beas et al, 2012; Bhandari et al, 2015; Denker et al, 1996; Garcia-Marcos et al, 2011; Garcia-Marcos et al, 2010; Garcia-Marcos et al, 2009; Ghosh et al, 2008; Lin et al, 2011; Lo et al, 2015; Lopez-Sanchez et al, 2013; Lopez-Sanchez et al, 2015a; Ma et al, 2015d) GIV-myc in pcDNA 3.1 (Bhandari et al, 2015)
GIV fragments	In the following vector backbones: pET28b (His-tagged, bacterial), pGEX-4T (GST-tagged, bacterial), pCEFL-GST (GST-tagged, mammalian), or pcDNA3.1-CFP (CFP-tagged, mammalian)	N-terminally placed His6-, GST- or CFP- tag.	Placement of each tag in the C-terminus significantly reduces binding of GIV-CT to G protein (reason unclear).	pER28b (ADDGENE ID# 69864) (Bhandari et al, 2015; Garcia-Marcos et al, 2011; Garcia-Marcos et al, 2010; Garcia-Marcos et al, 2009) pGEX-4T (ADDGENE ID# 69863) (Garcia-Marcos et al, 2011; Garcia-Marcos et al, 2010; Ogier-Denis et al, 1996) pCEFL (Bhandari et al, 2015; Lo et al, 2015; Midde et al, 2015) pcDNA3.1 CFP (ADDGENE ID# 69532) (Midde et , 2015)