pRcCMV Cep215 (Nigg CW493)
(Plasmid #41152)

• Depositing Lab: Erich Nigg
• Publication: Graser et al J Cell Sci. 2007 Dec 15;120(Pt 24):4321-31. Epub 2007 Nov 27.

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

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).
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.