pcDNA3 mHIF-2α MYC (P405A/P530V/N851A)
(Plasmid #44027)

• Depositing Lab: Celeste Simon
• Publication: Hu et al Mol Biol Cell. 2007 Nov;18(11):4528-42. Epub 2007 Sep 5.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5446
• Total vector size (bp): 8070
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mHIF-2α (P405A/P530V/N851A)
• Alt name: HIF-2α
• Alt name: Hypoxia-inducible factor-2alpha
• Alt name: Epas1
• Species: M. musculus (mouse)
• Insert Size (bp): 2600
• Mutation: P405A, P530V, N851A
• GenBank ID: AK158430.1 AAH57870.1
• Entrez Gene: Epas1 (a.k.a. HIF-2alpha, HIF2A, HLF, HRF, MOP2, bHLHe73)
• Promoter: CMV
• Tag / Fusion Protein:
  • Myc (C terminal on insert)

Cloning Information

• 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

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

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