Kif3A-EGFP-SNAP
(Plasmid #229852)

• Purpose: Kif3A (aa1-342)-Kif1A neck linker(17aa) fused to Kin1 coiled coil (aa345-406)-EGFP-SNAP-his. This Kif3A was used to link an oligo to the motor using the SNAP tag for single molecule experiments
• Depositing Lab: William Hancock
• Publication: DNA tensiometer reveals catch-bond detachment kinetics of kinesin-1, -2 and -3 (unpublished)

Backbone

• Vector backbone: pet28
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Used BL21 (DE3) E.coli for protein expression. Grew at 37 degrees for 3-4 hours until inducing with IPTG at an OD ~1, then grew overnight at 21C before harvesting
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Kif3A
• Alt name: Kif3A-Kif3ANL-Kin1CC
• Species: M. musculus (mouse)
• Insert Size (bp): 2574
• Entrez Gene: Kif3a (a.k.a. Kif3, Kifl, Kns3)
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • 6xHis
  • SNAP-tag
  • EGFP

Cloning Information

• Cloning method: Gene Synthesis
• 5′ sequencing primer: T7 promoter

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Genescript

Terms and Licenses

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