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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 | 11941 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneEGFP-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 3970
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 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)990
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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 EGFP-N (CGTCGCCGTCCAGCTCGACCAG) (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.
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:
Ub-G76V-GFP was a gift from Nico Dantuma (Addgene plasmid # 11941 ; http://n2t.net/addgene:11941 ; RRID:Addgene_11941) -
For your References section:
Short-lived green fluorescent proteins for quantifying ubiquitin/proteasome-dependent proteolysis in living cells. Dantuma NP, Lindsten K, Glas R, Jellne M, Masucci MG. Nat Biotechnol. 2000 May . 18(5):538-43. 10.1038/75406 PubMed 10802622