pcDNA3 mHIF-2α MYC (P405A/P530V/N851A)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||44027||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5446
- Total vector size (bp) 8070
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namemHIF-2α (P405A/P530V/N851A)
Alt nameHypoxia-inducible factor-2alpha
SpeciesM. musculus (mouse)
Insert Size (bp)2600
MutationP405A, P530V, N851A
GenBank IDAK158430.1 AAH57870.1
Entrez GeneEpas1 (a.k.a. HIF-2alpha, HIF2A, HLF, HRF, MOP2, bHLHe73)
- Promoter CMV
/ Fusion Protein
- Myc (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site KpnI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer CMV-F; T7
- 3′ sequencing primer BGH-rev; Sp6 (Common Sequencing Primers)
This plasmid encodes an oxygen-regulation insensitive (normoxia-stable and active) full-length murine HIF-2α.
Full-length murine HIF-2α cDNA was generated by reverse transcription-PCR and inserted into a pcDNA3 expression vector by using primers incorporated with restriction sites.
A 10-amino-acid c-myc tag was further inserted at the C terminus of the HIF-2α cDNAs by using a Pfu-PCR-based protocol. The same Pfu-PCR protocol was then used to change residues Proline 405, Proline 530 and Asparagine 851. The newly constructed plasmid was sequenced to confirm encoding of the correct protein.
This plasmid has an A94V polymorphism that does not affect protein function.
Alternate plasmid names: HIF-2α triple mutant (TM); pcDNA3 HIF-2α TM
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:pcDNA3 mHIF-2α MYC (P405A/P530V/N851A) was a gift from Celeste Simon (Addgene plasmid # 44027 ; http://n2t.net/addgene:44027 ; RRID:Addgene_44027)
For your References section:The N-terminal transactivation domain confers target gene specificity of hypoxia-inducible factors HIF-1alpha and HIF-2alpha. Hu CJ, Sataur A, Wang L, Chen H, Simon MC. Mol Biol Cell. 2007 Nov;18(11):4528-42. Epub 2007 Sep 5. 10.1091/mbc.e06-05-0419 PubMed 17804822