Addgene - pYES2-Ub-R-GFP Plasmid Data

Plasmid 11953: pYES2-Ub-R-GFP

Gene/insert name:
Ub-R-GFP
Alt name:
Ubiquitin
Alt name:
Ub
Insert size:
998
Mutation:
Ubiquitin fused to N-terminus of GFP. Arginine at position 1 between Ub and GFP (see article 10802622). N-end rule degradation signal.
Vector backbone:
pYES2
(Search Vector Database)
Backbone manufacturer:
Invitrogen
Vector type:
Yeast Expression
Backbone size w/o insert (bp):
5830
Cloning site 5':
EcoRI
Site destroyed during cloning:
No
Cloning site 3':
NotI
Site destroyed during cloning:
No
5' sequencing primer:
GAL1 primer List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
URA3
Sequence: View sequences (2)
Principal Investigator:
Nico Dantuma
Terms and Licenses:
MTA

Comments:

The ubiquitin open reading frame was amplified by PCR from the Ub-Pro-Gal plasmid with the sense primer 5'-GCG GAATTCACCATGCAGATCTTCGTGAAGACT-3' and the antisense primer 5'-GCG GGATCCTGTCGACCAAGCTTCCCXXX CCCACCTCTGAGACGGAGTAC-3' where the XXX leads to a arginine at position 1 (see article 10802622, Figure 1). The PCR product was cloned into the EcoRI and BamHI sites of the EGFP-N1 vector from Clontech.

The Ub-R-GFP insert was then excised with EcoRI and NotI and cloned into pYES2.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Inhibition of ubiquitin/proteasome-dependent proteolysis in Saccharomyces cerevisiae by a Gly-Ala repeat. Heessen et al (FEBS Lett. 2003 Dec 4. 555(2):397-404. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 11953" in your Materials and Methods section.

Price: $65.00

Available to academic and non-profits only

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This is commonly requested with

11952: pYES2-Ub-M-GFP

11954: pYES2-Ub-G76V-GFP

11939: Ub-R-GFP

11941: Ub-G76V-GFP

11938: Ub-M-GFP