Tol2-LyzC-RacFRET-U6ac-ctrl guides
(Plasmid #168247)

• Purpose: "label active Rac; control for neutrophil-specific knockout"
• Depositing Lab: Qing Deng
• Publication: Wang et al J Cell Sci. 2021 Apr 15;134(8). pii: 237799. doi: 10.1242/jcs.258574. Epub 2021 Apr 22.

Backbone

• Vector backbone: pME-GFP
• Modifications to backbone: remove GFP and add RacFRET
• Vector type: Zebrafish expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: RacFRET
• gRNA/shRNA sequence: gggccaatgcgagaccctgt/gggctgtatcccagagtccc
• Promoter: LyzC

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: GGTCGACTCGCCACCATGGTGAGCAAGGGCGAGG
• 3′ sequencing primer: CTCCACCGCGGCCGCTTACATAATTACACACTTTGTC

Resource Information

• Supplemental Documents:
  • lyzC-GFP-pa-U6CA-ctrlg-1101.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Reference for the RacFRET (Raichu-Rac1) element:
D. L. Graham, P. N. Lowe, and P. A. Chalk. A method to measure the interaction of Rac/Cdc42 with their binding partners using fluorescence resonance energy transfer between mutants of green fluorescent protein. Anal.Biochem. 296:208-217, 2001.