pEB1N-LZ-LOV2(fast)
(Plasmid #107690)

• Purpose: N-terminal half of π-EB1 with fast LOV2, no fluorescent tag
• Depositing Lab: Torsten Wittmann
• Publication: van Haren et al Nat Cell Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 10.1038/s41556-017-0028-5.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3941
• Total vector size (bp): 5078
• 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): 1137
• Mutation: EB1 aa 1-185; resistant to EB1 shRNA#3 (Addgene #37927)
• GenBank ID: NM_012325.2
• Entrez Gene: MAPRE1 (a.k.a. EB1)
• Promoter: CMV
• Tags / Fusion Proteins:
  • GCN4 leucine zipper (C terminal on insert)
  • A. sativa phototropin 1 LOV2 domain (I427V, H519L) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nhe1 (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: SV40 reverse

Resource Information

• Supplemental Documents:
  • 107690-EB1-N-LZ-LOV2 fast
• A portion of this plasmid was derived from a plasmid made by: EB1 from L. Cassimeris (Addgene #17234) LOV2 from Klaus Hahn (Addgene #22027)
• Articles Citing this Plasmid:
  • 2 References

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) and mutations I427V and H519L resulting in faster dark state recovery.

This LOV2 domain has the following mutations: I427V, H519L. The I427V mutation increases the dark state reversion rate of the LOV2 domain. Zdk1 binds efficiently to this LOV2 domain.