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Addgene

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

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 107690 Standard format: Plasmid sent in bacteria as agar stab 1 $85

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
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • 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

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

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.

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pEB1N-LZ-LOV2(fast) was a gift from Torsten Wittmann (Addgene plasmid # 107690 ; http://n2t.net/addgene:107690 ; RRID:Addgene_107690)
  • For your References section:

    Local control of intracellular microtubule dynamics by EB1 photodissociation. van Haren J, Charafeddine RA, Ettinger A, Wang H, Hahn KM, Wittmann T. Nat Cell Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 10.1038/s41556-017-0028-5. 10.1038/s41556-017-0028-5 [pii] PubMed 29379139