pGEX6P1-optimizedNedd4.2F823D
(Plasmid
#165105)
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Purposebacterial expression of a GST fusion of Human Nedd4.2d F823D
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 165105 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepGEX6P1
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Backbone manufacturerGE Healthcare
- Backbone size w/o insert (bp) 4900
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Vector typeBacterial Expression
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 nameNedd4.2d F823D
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Alt nameNEDD4L
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SpeciesH. sapiens (human)
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Insert Size (bp)2868
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Mutationcodon optimized for E. coli expression; F823D; mutation for destabilization of trimer formation
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Entrez GeneNEDD4L (a.k.a. NEDD4-2, NEDD4.2, PVNH7, RSP5, hNEDD4-2)
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Tag
/ Fusion Protein
- GST from plasmid. (N terminal on insert)
Cloning Information
- Cloning method Ligation Independent Cloning
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
Best yields for protein purification were found in BL21 (DE) cells using an overnight induction at 16 degrees C.
5' Cloning Site: BamHI (not destroyed), 3' Cloning Site: EcoRI (not destroyed).
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:
pGEX6P1-optimizedNedd4.2F823D was a gift from Arthur Haas (Addgene plasmid # 165105 ; http://n2t.net/addgene:165105 ; RRID:Addgene_165105) -
For your References section:
Oligomerization of the HECT ubiquitin ligase NEDD4-2/NEDD4L is essential for polyubiquitin chain assembly. Todaro DR, Augustus-Wallace AC, Klein JM, Haas AL. J Biol Chem. 2018 Nov 23;293(47):18192-18206. doi: 10.1074/jbc.RA118.003716. Epub 2018 Oct 4. 10.1074/jbc.RA118.003716 PubMed 30287686