EMTB-mCherry
(Plasmid #26742)

• Depositing Lab: William Bement
• Depositing Organization: University of Wisconsin, Madison
  Located in the United States of America
• Publication: Miller et al Nat Cell Biol. 2009 Jan . 11(1):71-7.
• Citations: 14 Articles

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: Microtubule binding-domain of ensconsin (amino acids 18-282 of GenBank reference sequence X73882)
• GenBank ID: X73882
• Entrez Gene: MAP7 (a.k.a. E-MAP-115, EMAP115)
• Tags / Fusion Proteins:
  • 2x mCherry (N terminal on backbone)
  • Myc (N terminal on insert)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: A EMTB construct was a gift from Chloë Bulinski (Columbia University)

Terms and Licenses

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

Depositor Comments

Note: It is difficult to sequence entirely through 2 tandem mCherrys. Users are encouraged to perform a double-digest on this plasmid with BamHI and EcoRI to verify the presence of a ~2.2kb band corresponding to the EMTB insert + 2XmCherry. Please see above attachment.

The microtubule binding domain of ensconsin (EMTB) was fused to multiple mCherry molecules in order to visualize microtubules. Please note that this plasmid has recombined and is currently only 2X mCherry.

mCherry sequences have point mutation at R154 (CGG to AGA) for optimal codon usage in Xenopus.

DNA (Catalog # 26742-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 26742-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.