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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 11954 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepYES2
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5830
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Vector typeYeast Expression
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Selectable markersURA3
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameUb-G76V-GFP
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Alt nameUbiquitin
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Alt nameUb
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Insert Size (bp)998
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MutationUbiquitin fused to N-terminus of GFP. Glycine 76 mutated to Valine.
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer GAL1 primer (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor 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'-GCGGGATCCTGTCGACCAAGCTTC
CCCACCACACCTCTGAGACGGAGTAC-3'. The PCR product was cloned into the EcoRI and BamHI sites of the EGFP-N1 vector from Clontech.
The Ub-G76V-GFP insert was then excised with EcoRI and NotI and cloned into pYES2.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pYES2-Ub-G76V-GFP was a gift from Nico Dantuma (Addgene plasmid # 11954 ; http://n2t.net/addgene:11954 ; RRID:Addgene_11954) -
For your References section:
Inhibition of ubiquitin/proteasome-dependent proteolysis in Saccharomyces cerevisiae by a Gly-Ala repeat. Heessen S, Dantuma NP, Tessarz P, Jellne M, Masucci MG. FEBS Lett. 2003 Dec 4. 555(2):397-404. 10.1016/S0014-5793(03)01296-1 PubMed 14644450