pEB1N-mApple-LZ-LOV2
(Plasmid #197418)

• Purpose: Mammalian expression of the mApple-tagged N-terminal half of photo-inactivated π-EB1
• Depositing Lab: Torsten Wittmann
• Publication: Dema et al Curr Biol. 2022 Mar 14;32(5):1197-1205.e4. doi: 10.1016/j.cub.2022.01.017. Epub 2022 Jan 31.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3936
• Total vector size (bp): 5798
• Modifications to backbone: removed EGFP
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: MAPRE1
• Alt name: EB1
• Species: H. sapiens (human)
• Insert Size (bp): 1857
• Mutation: EB1 aa 1-185; resistant to EB1 shRNA#3 (Addgene #37927)
• GenBank ID: NM_012325.2
• Entrez Gene: MAPRE1 (a.k.a. EB1)
• Tags / Fusion Proteins:
  • GCN4 leucine zipper (C terminal on insert)
  • mApple (C terminal on insert)
  • A. sativa phototropin 1 LOV2 domain (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: SV40 reverse

Resource Information

• Supplemental Documents:
  • pEB1N-mApple-LZ-LOV2.gbk
• A portion of this plasmid was derived from a plasmid made by: EB1 from L. Cassimeris (Addgene #17234) LOV2 from Klaus Hahn (Addgene #22027)

Terms and Licenses

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

Depositor Comments

LOV2 domain starts with LAAA reported to stabilize Jα helix docking (Strickland et al. PMID: 22388287)