RGD-sfGFP-I27-CC
(Plasmid #85392)

• Purpose: MTFM sensor to measure integrin mechanical forces. Integrin binding domain fused to titan immunoglobulin domain (I27) with two cysteines (CC) for immobilization and sfGFP for fluorescent labeling
• Depositing Lab: Khalid Salaita
• Publication: Galior et al Nano Lett. 2016 Jan 13;16(1):341-8. doi: 10.1021/acs.nanolett.5b03888. Epub 2015 Dec 7.

Backbone

• Vector backbone: pET-22b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5493
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use BL21(DE3) strain for expression
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: RGD motif for integrin binding fused to sfGFP and titin immunoglobulin domain (I27)
• Alt name: KS27-ECC
• Promoter: T7
• Tag / Fusion Protein:
  • 6x His (C terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7-term

Resource Information

• Supplemental Documents:
  • 85392.gb

Terms and Licenses

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

Depositor Comments

Fibronectin binding motif (RGD) is TVYAVTGRGDSPASSAA.

Superfolder green fluorescent protein (sfGFP) is present at the N-terminus of I27 domain.

I27 domain is the 27th immunoglobulin domain of cardiac titin. Two cysteine residues were added at the C-terminus of I27 to allow immobilization onto a gold nanoparticle.

Please see the associated publication for material and methods information.