EcpH-TM
(Plasmid #85389)

• Purpose: Expresses ecliptic pHluorin–ICAM-1 (intercellular adhesion molecule 1) in mammalian cells. Incorporates into viral membrane to label pseudoviruses
• Depositing Lab: Gregory Melikyan
• Publication: Miyauchi et al Biochem J. 2011 Mar 15;434(3):559-69. doi: 10.1042/BJ20101588.

Backbone

• Vector backbone: p3xFLAG CMV9
• Backbone manufacturer: Sigma
• Backbone size w/o insert (bp): 6373
• 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: ecliptic pHluorin (EcpH) fused to N-terminus of human ICAM-1 transmembrane domain
• Species: Synthetic
• Insert Size (bp): 900
• Promoter: CMV
• Tag / Fusion Protein:
  • 3x FLAG (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: hGH-pA-R

Terms and Licenses

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

Depositor Comments

A PCR product containing the transmembrane domain of human ICAM-1 was amplified from pCDM8 ICAM-1 (Addgene plasmid # 8632). ecliptic pHluorin (EcpH) was amplified from a Gag-EcpH-expression vector (Sherer et al., Traffic 2003). The EcpH fragment was cloned into p3xFLAG CMV9 using HindIII and EcoRI sites and the ICAM-1 TM fragment was cloned using EcoRI and BamHI.