pcDNA3.1-KozATG-dH6.2-2XG4S-actin
(Plasmid #73263)

• Purpose: Expresses dH6.2-actin fusion within mammalian cells. (dH6.2, FAP)
• Depositing Lab: Marcel Bruchez
• Depositing Organization: Carnegie Mellon University
  Located in United States of America
• Publication: Yan et al Chemphyschem. 2014 Mar 17;15(4):687-95. doi: 10.1002/cphc.201300757. Epub 2013 Nov 5.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5382
• Total vector size (bp): 7458
• Modifications to backbone: The NdeI site was removed from the CMV promoter
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: No special instructions.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: KozATG-dH6.2-2XG4S-actin
• Alt name: dH6-actin
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 2070
• Mutation: The dH6.2 FAP has one cysteine in each domain changed to an alanine
• Entrez Gene: ACTB (a.k.a. BKRNS, BNS, BRWS1, CSMH, DDS1, PS1TP5BP1, THC8)
• Promoter: CMV
• Tag / Fusion Protein:
  • The FAP and actin are fused with 2 copies of a G4S linker

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CMVF
• 3′ sequencing primer: BGHR

Terms and Licenses

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

Depositor Comments

Please visit the Molecular Biosensor and Imaging Center website for current protocols and information about dyes to be used in combination with the proteins expressed from this plasmid.

http://pathways.mbic.cmu.edu/

DNA (Catalog # 73263-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 # 73263-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.