BiP-mGFP P495L
(Plasmid #62232)

• Purpose: Mutant of hamster BiP-mGFP unable to bind clients.
• Depositing Lab: Erik Snapp
• Publication: Lai et al Mol Biol Cell. 2010 Jun 15;21(12):1909-21. doi: 10.1091/mbc.E09-12-1066. Epub 2010 Apr 21.

Backbone

• Vector backbone: clontech N1
• Backbone manufacturer: clontech
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 6700
• Vector type: Mammalian Expression
• Selectable markers: Gentamicin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hamster BiP-mGFP-KDEL
• Species: Synthetic; hamster
• Insert Size (bp): 2702
• Promoter: cmv
• Tag / Fusion Protein:
  • mGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: notI (not destroyed)
• 5′ sequencing primer: tggtttagtgaaccgt
• 3′ sequencing primer: GTTCAGGGGGAGGTGTG

Terms and Licenses

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

Depositor Comments

Avoid overexpression. Low expressing cells are best for imaging. High expression leads to protein aggregation in the nuclear envelope. See supplementary data for original paper.
This mutant can not bind client proteins.
Do NOT use this as an ER marker. BiP is a functional protein and significantly impacts the ER environment increasing stress resistance. Use ER-GFP if you simply want a marker for the ER when imaging cells.