pcDNA3-PKCsub-YPet-NES
(Plasmid
#138207)
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PurposeEncodes C-terminal (substrate) fragment of FRET-based bimolecular PKC activity reporter (bimCKAR); cytosol targeted. Use in conjunction with pcDNA3-Cerulean-FHA1-NES
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 138207 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5472
- Total vector size (bp) 6294
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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 namePKCsub-YPet-NES
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SpeciesSynthetic
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Insert Size (bp)822
- Promoter CMV
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Tags
/ Fusion Proteins
- YPet (C terminal on insert)
- Nuclear export signal (NES) (C terminal on insert)
- 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 XbaI (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-PKCsub-YPet-NES was a gift from Jin Zhang (Addgene plasmid # 138207 ; http://n2t.net/addgene:138207 ; RRID:Addgene_138207) -
For your References section:
Luminescent kinase activity biosensors based on a versatile bimolecular switch. Herbst KJ, Allen MD, Zhang J. J Am Chem Soc. 2011 Apr 20;133(15):5676-9. doi: 10.1021/ja1117396. Epub 2011 Mar 25. 10.1021/ja1117396 PubMed 21438554