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PurposePlasma membrane-targeted EKAR4.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 174440 | 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) 7518
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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 namepmEKAR4
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Alt nameEKAR4-Kras
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Insert Size (bp)2046
- Promoter CMV
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Tags
/ Fusion Proteins
- 6xHIS - T7 tag (gene 10 leader) - Xpress (TM) tag
- C-terminal targeting sequence from Kras (C terminal on insert)
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.
Depositor Comments
Please note there is a single ambiguous base in the insert that we could not resolve via NGS. Depositor has confirmed this to be a C as expected via sanger sequencing.
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-pmEKAR4 was a gift from Jin Zhang (Addgene plasmid # 174440 ; http://n2t.net/addgene:174440 ; RRID:Addgene_174440) -
For your References section:
Signaling diversity enabled by Rap1-regulated plasma membrane ERK with distinct temporal dynamics. Keyes J, Ganesan A, Molinar-Inglis O, Hamidzadeh A, Zhang J, Ling M, Trejo J, Levchenko A, Zhang J. eLife. 2020 May 26;9. pii: 57410. doi: 10.7554/eLife.57410. 10.7554/eLife.57410 PubMed 32452765