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pcDNA3 mHIF-1α MYC (P402A/P577A/N813A)
(Plasmid #44028)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 44028 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5446
  • Total vector size (bp) 7950
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    mHIF-1α (P402A/P577A/N813A)
  • Alt name
    HIF-1α
  • Alt name
    Hypoxia inducible factor 1, alpha subunit
  • Alt name
    HIF1alpha
  • Species
    M. musculus (mouse)
  • Insert Size (bp)
    2500
  • Mutation
    P402A, P577A, N813A
  • GenBank ID
    NM_010431.2 NP_034561.2
  • Entrez Gene
    Hif1a (a.k.a. AA959795, HIF-1-alpha, HIF1-alpha, HIF1alpha, MOP1, bHLHe78)
  • Promoter CMV
  • Tag / Fusion Protein
    • Myc (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site Unknown (unknown if destroyed)
  • 5′ sequencing primer CMV-F; T7
  • 3′ sequencing primer BGH-rev; Sp6
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

This plasmid encodes an oxygen-regulation insensitive (normoxia-stable and active) full-length murine HIF-1α.

Full-length murine HIF-1α 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-1α cDNAs by using a Pfu-PCR-based protocol. The same Pfu-PCR protocol was then used to change residues Proline 402, Proline 577 and Asparagine 813 to alanines. The newly constructed plasmid was sequenced to confirm encoding of the correct protein.

Alternate plasmid names: HIF-1α triple mutant (TM); pcDNA3 HIF-1α TM

How to cite this plasmid ( Back to top)

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-1α MYC (P402A/P577A/N813A) was a gift from Celeste Simon (Addgene plasmid # 44028)
  • 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