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Yeplac181

Manufactured by Thermo Fisher Scientific
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YEplac181 is a yeast expression plasmid that can be used for the expression of recombinant proteins in Saccharomyces cerevisiae. It contains a 2-micron origin of replication, the URA3 selectable marker, and a multi-cloning site for the insertion of target genes.

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Lab products found in correlation

Pqe30, by Qiagen (2 mentions) Pyes2, by Thermo Fisher Scientific (2 mentions)

2 protocols using yeplac181

1

Ubiquitin Constructs for Yeast and E. coli

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For expression in yeast, mycUBB+1, mycUbG76V20aa and mycUbG76V20aa were amplified by PCR from plasmids provided by N. Dantuma and were cloned using Sac1 and Kpn1 into GAL4 or ADH promoter-carrying vectors, pYES2 (Invitrogen) and YEplac181 respectively. mycUb20, mycUb25, mycUbV70D,G76V25aa constructs were created by side-directed mutagenesis of plasmids described in preceding sentence.
For E. coli expression plasmids, open reading frames (ORFs) of UBB+1, UbG76V20aa, UbG76V20aa, Ub20, Ub25 were amplified by PCR from the corresponding yeast vectors described above, and ORFs of Rpn10, Rpn11, and Ubp6 were amplified from S. cerevisiae genomic DNA. All PCR products were cloned into pQE30 (QIAGEN) using Sac1 and Pst1.
Krutauz D., Reis N., Nakasone M.A., Siman P., Zhang D., Kirkpatrick D.S., Gygi S.P., Brik A., Fushman D, & Glickman M.H. (2014). Extended ubiquitin species are protein-based DUB inhibitors. Nature chemical biology, 10(8), 664-670.
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2

Ubiquitin Constructs for Yeast and E. coli

Check if the same lab product or an alternative is used in the 5 most similar protocols
For expression in yeast, mycUBB+1, mycUbG76V20aa and mycUbG76V20aa were amplified by PCR from plasmids provided by N. Dantuma and were cloned using Sac1 and Kpn1 into GAL4 or ADH promoter-carrying vectors, pYES2 (Invitrogen) and YEplac181 respectively. mycUb20, mycUb25, mycUbV70D,G76V25aa constructs were created by side-directed mutagenesis of plasmids described in preceding sentence.
For E. coli expression plasmids, open reading frames (ORFs) of UBB+1, UbG76V20aa, UbG76V20aa, Ub20, Ub25 were amplified by PCR from the corresponding yeast vectors described above, and ORFs of Rpn10, Rpn11, and Ubp6 were amplified from S. cerevisiae genomic DNA. All PCR products were cloned into pQE30 (QIAGEN) using Sac1 and Pst1.
Krutauz D., Reis N., Nakasone M.A., Siman P., Zhang D., Kirkpatrick D.S., Gygi S.P., Brik A., Fushman D, & Glickman M.H. (2014). Extended ubiquitin species are protein-based DUB inhibitors. Nature chemical biology, 10(8), 664-670.
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