pcDNA3.1(+)-RasAR(R89L)-NES
(Plasmid
#205568)
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PurposeRasAR negative-control mutant plasmid. Targeted to cytosol.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 205568 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3.1(+)
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5363
- Total vector size (bp) 7151
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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)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 nameRasAR(R89L)-NES
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Alt nameCerulean3-RBD(Raf1 51-131, R89L)-pseudoligand-YPet-NES
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SpeciesH. sapiens (human), Synthetic
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Insert Size (bp)1788
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MutationArg 89 mutated to Leu in Raf1 RBD region.
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Entrez GeneRAF1 (a.k.a. CMD1NN, CRAF, NS5, Raf-1, c-Raf)
- Promoter CMV
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Tag
/ Fusion Protein
- Nuclear export signal (NES) (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer CMV forward
- 3′ sequencing primer BGH reverse (Common Sequencing Primers)
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.
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:
pcDNA3.1(+)-RasAR(R89L)-NES was a gift from Jin Zhang (Addgene plasmid # 205568 ; http://n2t.net/addgene:205568 ; RRID:Addgene_205568) -
For your References section:
Fluorescent Biosensor for Measuring Ras Activity in Living Cells. Weeks R, Zhou X, Yuan TL, Zhang J. J Am Chem Soc. 2022 Sep 28;144(38):17432-17440. doi: 10.1021/jacs.2c05203. Epub 2022 Sep 19. 10.1021/jacs.2c05203 PubMed 36122391