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Purposemammalian expression of GFP-Kif5c
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Depositing Lab
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Depositing OrganizationUniversity of Leeds
Located in United Kingdom of Great Britain and Northern Ireland -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 71853 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 71853-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 71853-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFPC3
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4700
- Total vector size (bp) 7700
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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)Kanamycin, 50 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 nameKif5c
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Alt namekinesin family member 5C
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SpeciesR. norvegicus (rat)
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Insert Size (bp)3000
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MutationKif5c starts at AA3
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Entrez GeneKif5c
- Promoter CMV
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Tag
/ Fusion Protein
- EGFP (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer EGFP-C
- 3′ sequencing primer SV40pA-R
- (Common Sequencing Primers)
Resource Information
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
Following transfection, look at cells that have low levels of expression (for the wild type Kif5c protein) to investigate dynamic behaviour, as high levels of expression result in bundling of microtubules.
DNA (Catalog # 71853-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 # 71853-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:
pGFP-Kif5c was a gift from Michelle Peckham (Addgene plasmid # 71853 ; http://n2t.net/addgene:71853 ; RRID:Addgene_71853) -
For your References section:
Differential trafficking of Kif5c on tyrosinated and detyrosinated microtubules in live cells. Dunn S, Morrison EE, Liverpool TB, Molina-Paris C, Cross RA, Alonso MC, Peckham M. J Cell Sci. 2008 Apr 1;121(Pt 7):1085-95. doi: 10.1242/jcs.026492. Epub 2008 Mar 11. 10.1242/jcs.026492 PubMed 18334549
Map uploaded by the depositor.
