pEB1N-mScarlet-LZ-LOV2
(Plasmid #197419)

• Purpose: Mammalian expression of the mScarlet-tagged N-terminal half of photo-inactivated π-EB1
• Depositing Lab: Torsten Wittmann
• Publication: Dema et al Elife. 2023 Jan 30;12:e84143. doi: 10.7554/eLife.84143.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3941
• Total vector size (bp): 5786
• 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): 1845
• 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)
  • mScarlet-I (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-mScarlet-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).

Please visit https://www.biorxiv.org/content/10.1101/2022.09.22.509085v1 for bioRxiv preprint.