human EB1 in PET28a for protein expression
(Plasmid #39297)

• Depositing Labs: Tim Mitchison, Jennifer Tirnauer
• Publication: Tirnauer lab EB1 plasmids (unpublished)

Backbone

• Vector backbone: PET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 6173
• Modifications to backbone: none
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Can be propagated in DH5alpha. For protein expression, grow in BL21 pLysS strain. Induce with IPTG about 3 hours and purify on Nickel agarose. For the full expression and purification protocol see Tirnauer et al MBoC 2002 EB1–Microtubule Interactions in Xenopus Egg Extracts: Role of EB1 in Microtubule Stabilization and Mechanisms of Targeting to Microtubules
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: EB1
• Alt name: MAPRE1
• Species: H. sapiens (human)
• Insert Size (bp): 804
• Entrez Gene: MAPRE1 (a.k.a. EB1)
• Promoter: T7
• Tag / Fusion Protein:
  • 6HIS (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: T7 promotor primer
• 3′ sequencing primer: T7 terminator primer

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Jen-Yu (technician in Barbara Bierer's lab)

Terms and Licenses

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

Depositor Comments

sequenced. See Berrueta et al 1999