pcDNA 3.3 ss-V5-hFZD4 (aa 37-537)
PurposeExpresses human FZD4 with N-terminal V5-tag in mammalian cells, contains artificial signal sequence upstream of tag.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||115787||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepcDNA3.3 TOPO TA
- Backbone size w/o insert (bp) 5407
- Total vector size (bp) 7047
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
SpeciesH. sapiens (human)
Insert Size (bp)1640
Entrez GeneFZD4 (a.k.a. CD344, EVR1, FEVR, FZD4S, Fz-4, Fz4, FzE4, GPCR, hFz4)
- Promoter CMV
/ Fusion Protein
- V5 (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI (not destroyed)
- 3′ cloning site AgeI (not destroyed)
- 5′ sequencing primer CGCAAATGGGCGGTAGGCGTG
- 3′ sequencing primer CTTCCGTGTTTCAGTTAGC (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byImage clone 5199771
Terms and Licenses
N-terminal V5 tag has no substantial effect in TOPFlash reporter assays induced with Norrin, compared to untagged control. TOPFlash reporter assay can be inhibited by transfection of too much of this plasmid (see Lai et al., 2017 for detailed description of transfection conditions).
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:pcDNA 3.3 ss-V5-hFZD4 (aa 37-537) was a gift from Harald Junge (Addgene plasmid # 115787 ; http://n2t.net/addgene:115787 ; RRID:Addgene_115787)
For your References section:TSPAN12 Is a Norrin Co-receptor that Amplifies Frizzled4 Ligand Selectivity and Signaling. Lai MB, Zhang C, Shi J, Johnson V, Khandan L, McVey J, Klymkowsky MW, Chen Z, Junge HJ. Cell Rep. 2017 Jun 27;19(13):2809-2822. doi: 10.1016/j.celrep.2017.06.004. 10.1016/j.celrep.2017.06.004 PubMed 28658627