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. Author manuscript; available in PMC: 2019 Mar 17.
Published in final edited form as: Nature. 2018 Sep 17;561(7724):561–564. doi: 10.1038/s41586-018-0526-z

Extended Data Table 3. E. coli expression constructs for C. thermophilum proteins used in this study.

Name of construct Method of cloning Comments/experiments
pGEX4T2 Vps29 Gene was amplified from synthetic template (GenScript) and ligated using BamHI/Xhol restriction digestion sites protein crystallization
pGEX4T2psp Vps29 Introduced additional cleavage site for PreScission protease co-expression and purification of complex
pET28aH6 Vps35 Gene was amplified from synthetic template (GenScript) and ligated using Ncol/Xhol restriction digestion sites, His-tag introduced with amplification primers co-expression and purification of complex
pET28aH6 Vps26 Gene was amplified from synthetic template (GenScript) and ligated using Ncol/Xhol, His-tag introduced with amplification primers co-expression and purification of complex
pGEX4T2 Vps5 Gene was amplified from synthetic template (GenScript) and ligated using BamHI/Xhol liposome pelleting
pRSFDuetH6 Vps5 Gene was amplified from synthetic template (GenScript), cloned into MCS1, Ncol dimerization, complex purification.