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pRcCMV Cep215 (Nigg CW493)
(Plasmid #41152)

Ordering

This material is available to academics and nonprofits only.
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

  • Vector backbone
    pCJW206 (modified pEGFP-C1)
  • Backbone manufacturer
    Modified from Clontech
  • Backbone size w/o insert (bp) 4110
  • Total vector size (bp) 9800
  • Modifications to backbone
    FseI- 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
  • Vector type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Cep215
  • Alt name
    CDK5RAP2
  • Alt name
    CDK5 regulatory subunit associated protein 2
  • Alt name
    MCPH3
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    5700
  • GenBank ID
    AB046853
  • Entrez Gene
    CDK5RAP2 (a.k.a. C48, Cep215, MCPH3)
  • Promoter CMV
  • 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

  • A portion of this plasmid was derived from a plasmid made by
    Cep215 cDNA sequence from KIAA1633 clone (from Kazusa DNA Research Institute). 5' coding information from Marathon cDNA library (Clontech).

Terms and Licenses

  • Academic/Nonprofit Terms
  • 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.

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

  • 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.

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.

How to cite this plasmid ( Back to top)

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.

  • 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