pEF.myc.ER-E2-Crimson
(Plasmid #38770)

• Purpose: In vivo visualization of the endoplasmic reticulum (can be used for colocalization studies)
• Depositing Lab: Benjamin Glick
• Publication: Strack et al Biochemistry. 2009 Sep 8;48(35):8279-81.

Backbone

• Vector backbone: pEF/myc/ER
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5603
• Total vector size (bp): 6263
• 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): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: E2-Crimson
• Alt name: Modified DsRed-Express2
• Species: Synthetic
• Insert Size (bp): 675
• Promoter: EF-1a
• Tags / Fusion Proteins:
  • ER signal sequence (MGWSCIILFLVATATGAHS) (N terminal on backbone)
  • myc (C terminal on backbone)
  • ER retention signal (KDEL) (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: Unknown (unknown if destroyed)
• 5′ sequencing primer: EF1a_fwd
• 3′ sequencing primer: BGH_rev

Resource Information

• Supplemental Documents:
  • pEF.myc.ER-E2-Crimson Snapgene file
  • Annotated screen shot of N-term
• A portion of this plasmid was derived from a plasmid made by: Cloned by Birka Hein in the lab of Stefan Hell.
• Articles Citing this Plasmid:
  • 11 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Vector for targeting E2-Crimson to the ER in mammalian cells.

Substitutions in E2-Crimson relative to DsRed-Express2 are: E32V, Q66F, V71A, V73I, K83L, L85Q, F118L, L150N, I161N, K163M, V175C, Y193H, S197Y

IMPORTANT NOTE: There is an 82-bp intron after the ER signal sequence. The signal sequence will be in frame with the final product.