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Depositing Lab
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Depositing OrganizationUniversity of Basel
Located in Switzerland -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 41152 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 41152-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 41152-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCJW206 (modified pEGFP-C1)
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Backbone manufacturerModified from Clontech
- Backbone size w/o insert (bp) 4110
- Total vector size (bp) 9800
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Modifications to backboneFseI- and NotI-site-containing linker (AAT TCG GCC GGC CTA GCG GCC GCT GCA) inserted between EcoRI and PstI sites. Myc-tag-encoding fragment (aag ctt gct tac att tgc ttc tga cac aac tgt gtt cac tag caa cct caa aca gac acc atg gag cag aag ctg atc tcc gag gag gac ctg aac atg aat tc) inserted between HindIII and EcoRI sites. GFP-encoding sequence removed by excision with BspEI and AgeI and religation
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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 nameCep215
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Alt nameCDK5RAP2
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Alt nameCDK5 regulatory subunit associated protein 2
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Alt nameMCPH3
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SpeciesH. sapiens (human)
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Insert Size (bp)5700
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GenBank IDAB046853
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Entrez GeneCDK5RAP2 (a.k.a. C48, Cep215, MCPH3)
- Promoter CMV
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Tag
/ Fusion Protein
- Myc (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site FseI (unknown if destroyed)
- 3′ cloning site NotI (unknown if destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer SV40-pA-R
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byCep215 cDNA sequence from KIAA1633 clone (from Kazusa DNA Research Institute). 5' coding information from Marathon cDNA library (Clontech).
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
The Cep215 cDNA sequence was obtained from KIAA1633 clone (from Kazusa DNA Research Institute). Using PCR, a frameshift within the sequence was corrected and missing 5' coding information was obtained by PCR amplification from Marathon cDNA library (Clontech). Constructs were fused to yield the complete coding sequence of Cep215, and subcloned into a mammalian expression vector providing a Myc epitope-tag. All constructs were confirmed by sequencing.
DNA (Catalog # 41152-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 # 41152-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:
pRcCMV Cep215 (Nigg CW493) was a gift from Erich Nigg (Addgene plasmid # 41152 ; http://n2t.net/addgene:41152 ; RRID:Addgene_41152) -
For your References section:
Cep68 and Cep215 (Cdk5rap2) are required for centrosome cohesion. Graser S, Stierhof YD, Nigg EA. J Cell Sci. 2007 Dec 15;120(Pt 24):4321-31. Epub 2007 Nov 27. 10.1242/jcs.020248 PubMed 18042621