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PurposeImproved excitation-ratiometric biosensor for monitoring Akt/Protein Kinase B activity in living cells.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 184047 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3
- Backbone size w/o insert (bp) 5517
- Total vector size (bp) 6759
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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 nameExRai-AktAR2
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Alt nameAKTsubstrate-cpEGFP-FHA1
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Insert Size (bp)1242
- Promoter CMV
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Tag
/ Fusion Protein
- 6xHIS - T7 tag (gene 10 leader) - Xpress (TM) tag (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer CMV forward
- 3′ sequencing primer BGH reverse (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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-ExRai-AktAR2 was a gift from Jin Zhang (Addgene plasmid # 184047 ; http://n2t.net/addgene:184047 ; RRID:Addgene_184047) -
For your References section:
A Highly Sensitive Fluorescent Akt Biosensor Reveals Lysosome-Selective Regulation of Lipid Second Messengers and Kinase Activity. Chen M, Sun T, Zhong Y, Zhou X, Zhang J. ACS Cent Sci. 2021 Dec 22;7(12):2009-2020. doi: 10.1021/acscentsci.1c00919. Epub 2021 Dec 3. 10.1021/acscentsci.1c00919 PubMed 34963894