EMTB-3XGFP
(Plasmid #26741)

• Depositing Lab: William Bement
• Publication: Miller et al Nat Cell Biol. 2009 Jan . 11(1):71-7.

Backbone

• Vector backbone: pCS2+
• Backbone size w/o insert (bp): 4097
• Vector type: Mammalian Expression ; probe for microtubules

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ensconsin
• Alt name: E-MAP-115
• Alt name: EMTB
• Species: H. sapiens (human)
• Insert Size (bp): 816
• Mutation: Only contains the microtubule binding-domain of ensconsin (amino acids 18-283 of GenBank reference sequence X73882)
• GenBank ID: X73882
• Entrez Gene: MAP7 (a.k.a. E-MAP-115, EMAP115)
• Tags / Fusion Proteins:
  • Myc (N terminal on insert)
  • 3x EGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: T7

Resource Information

• Supplemental Documents:
  • Digest with EcoRI and XhoI
• A portion of this plasmid was derived from a plasmid made by: A EMTB construct was a gift from Chloë Bulinski (Columbia University)
• Articles Citing this Plasmid:
  • 29 References

Terms and Licenses

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

Depositor Comments

The microtubule binding domain of ensconsin (EMTB) was fused to 3 GFP molecules in order to visualize microtubules. For cloning details see Faire et al., 1999 Journal
of Cell Science 112, 4243-4255.

Note: It is difficult to sequence entirely through 3 tandem GFPs. Users are encouraged to perform a double-digest on this plasmid with EcoRI and XhoI to verify the presence of a ~3kb band corresponding to the EMTB insert + 3GFPs. Please see above attachment.