5HRE/GFP
(Plasmid #46926)

• Purpose: hypoxia-responsive enhanced green fluorescent protein (EGFP)-based system
• Depositing Labs: Martin Brown, Thomas Foster
• Publication: Vordermark et al Neoplasia. 2001 Nov-Dec;3(6):527-34.

Backbone

• Vector backbone: pEF/myc/cyto
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• 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: Unknown

Gene/Insert

• Gene/Insert name: 5X HRE of VEGF
• Alt name: VEGFA
• Alt name: VEGF
• Species: H. sapiens (human)
• Mutation: five copies of a 35-bp fragment from the hypoxia-responsive element (HRE) of the human VEGF gene and a human cytomegalovirus minimal promoter
• Entrez Gene: VEGFA (a.k.a. L-VEGF, MVCD1, VEGF, VPF)
• Promoter: EF-1α promoter
• Tag / Fusion Protein:
  • d2EGFP (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: EF1a-F
• 3′ sequencing primer: BGHrev

Resource Information

• Articles Citing this Plasmid:
  • 20 References

Terms and Licenses

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

Depositor Comments

The 35-bp human VEGF HRE sequence is: CCACAGTGCATACGTGGGCTCCAACAGGTCCTCTT

The d2EGFP fragment was amplified from a Clontech (Palo Alto, CA) vector and encodes a destabilized red- shifted variant of wild-type GFP with an excitation maximum of 488 nm, an emission maximum of 507 nm, and a half-life of approximately 2 hours.