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Purposeexpresses human cofilin R21Q mutant fused to mRFP in mammalian cells
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Depositing Lab
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Depositing OrganizationColorado State University, Fort Collins
Located in United States of America -
Citations
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 51279 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 51279-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 51279-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepmRFP-N1
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Backbone manufacturerR.Y. Tsien, PNAS 2002 v99 pp7877–7882
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Vector typeMammalian Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert namecofilin 1
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Alt nameCFL1
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SpeciesH. sapiens (human)
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Insert Size (bp)500
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MutationR21Q, non-rod-forming mutation that can be used to track cofilin-actin rods in vivo without causing cofilin overexpression artifacts
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Entrez GeneCFL1 (a.k.a. CFL, HEL-S-15, cofilin)
- Promoter CMV
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Tag
/ Fusion Protein
- mRFP (C terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site XmaI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer mRFP1-R
- (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.
Depositor Comments
This cofilin plasmid contains some silent mutations to make it resistant to an siRNA used in the Bamburg lab. The mutations change the wildtype sequence (AAG TCT TCA ACG CCA GAG GAG) into AAG AGC TCA ACC CCA GAG GAG.
DNA (Catalog # 51279-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 51279-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
pmRFP-N1 human cofilin R21Q was a gift from James Bamburg (Addgene plasmid # 51279 ; http://n2t.net/addgene:51279 ; RRID:Addgene_51279) -
For your References section:
A genetically encoded reporter for real-time imaging of cofilin-actin rods in living neurons. Mi J, Shaw AE, Pak CW, Walsh KP, Minamide LS, Bernstein BW, Kuhn TB, Bamburg JR. PLoS One. 2013 Dec 31;8(12):e83609. doi: 10.1371/journal.pone.0083609. PubMed 24391794