-
Depositing Lab
-
Sequence Information
Full plasmid sequence is not available for this item.
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 41151 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backbonepEGFP-T7/C1 (modified pEGFP-C1)
-
Backbone manufacturerModified from Clontech
- Backbone size w/o insert (bp) 4700
- Total vector size (bp) 7400
-
Modifications to backboneFrom pEGFP-C1: T7 promoter inserted into the vector NheI site
-
Vector typeMammalian Expression
-
Selectable markersNeomycin (select with G418)
Growth in Bacteria
-
Bacterial Resistance(s)Kanamycin, 50 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert nameCep170
-
Alt nameCentrosomal protein 170kDa
-
Alt nameKIAA0470
-
Alt nameFAM68A
-
SpeciesH. sapiens (human)
-
Insert Size (bp)2100
-
MutationContains C-terminal fragment (aa 755-1460, from XmnI site to the end of cDNA),
-
GenBank IDAB007939
-
Entrez GeneCEP170 (a.k.a. FAM68A, KAB, KIAA0470)
- Promoter CMV, T7
-
Tag
/ Fusion Protein
- eGFP (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BspEI (destroyed during cloning)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer EGFP-C; CMV-F
- 3′ sequencing primer SV40-pA-R (Common Sequencing Primers)
Resource Information
-
A portion of this plasmid was derived from a plasmid made byKIAA0470 cDNA obtained from the Kazusa DNA Research Institute
-
Article Citing this Plasmid
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
The KIAA0470 cDNA was obtained from the Kazusa DNA Research Institute. It was subcloned into pT7-EGFP-C1 (modified from pEGFP-C1; BD Biosciences Clontech), after removing the ATG. Specifically, a 5kb SalI-SnaBI (Klenow blunted) fragment from pBS/KIAA0470Δ5'UTR was ligated into pT7-EGFP-C1 (Nigg COM81) digested with XhoI (Klenow blunted). The 3' XhoI site was recreated. This resulted in Addgene plasmid #41150: pEGFP Cep170 (Nigg PID200). The plasmid was then digested with BspEI and XmnI, blunted, and religated to recover a plasmid containing only the Cep170 C-terminal portion (aa 755-1460, from XmnI site to the end of cDNA).
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:
pEGFP Cep170 C-term (Nigg GG2) was a gift from Erich Nigg (Addgene plasmid # 41151 ; http://n2t.net/addgene:41151 ; RRID:Addgene_41151) -
For your References section:
The forkhead-associated domain protein Cep170 interacts with Polo-like kinase 1 and serves as a marker for mature centrioles. Guarguaglini G, Duncan PI, Stierhof YD, Holmstrom T, Duensing S, Nigg EA. Mol Biol Cell. 2005 Mar;16(3):1095-107. Epub 2004 Dec 22. 10.1091/mbc.e04-10-0939 PubMed 15616186
Map uploaded by the depositor.