RGD-I27-CC
(Plasmid
#85393)
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PurposeMTFM mechanosensor for integrin. Integrin binding domain fused to titan immunoglobulin domain (I27) with two cysteines for immobilization and site for p-azidophenylalanine incorporation & Cy3 labeling
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 85393 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET-22b
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5493
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature30°C
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Growth Strain(s)DH5alpha
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Growth instructionsUse BL21(DE3) strain for expression
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameRGD motif for integrin binding fused to titin immunoglobulin domain (I27), containing a TAG (amber) codon
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Alt nameKS29-ECC
- Promoter T7
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Tag
/ Fusion Protein
- 6x His (C terminal on backbone)
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer T7
- 3′ sequencing primer T7-term (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Fibronectin binding motif (RGD) is TVYAVTGRGDSPASSAA.
A TAG (amber) codon was added between the RGD motif and I27 for labeling with p-azidophenylalanine, an unnatural amino acid. The RGD-I27-CC fusion protein can be labeled with the Cy3 dye at this site using Cu-free click cyanoaddition coupling.
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
RGD-I27-CC was a gift from Khalid Salaita (Addgene plasmid # 85393 ; http://n2t.net/addgene:85393 ; RRID:Addgene_85393) -
For your References section:
Titin-Based Nanoparticle Tension Sensors Map High-Magnitude Integrin Forces within Focal Adhesions. Galior K, Liu Y, Yehl K, Vivek S, Salaita K. Nano Lett. 2016 Jan 13;16(1):341-8. doi: 10.1021/acs.nanolett.5b03888. Epub 2015 Dec 7. 10.1021/acs.nanolett.5b03888 PubMed 26598972