G protein alpha-s-CFP
(Plasmid #55793)

• Purpose: G protein alpha-s internally tagged with ECFP and EE epitope
• Depositing Lab: Catherine Berlot
• Depositing Organization: Geisinger Health Center
  Located in United States of America
• Publication: Hynes et al J Biol Chem. 2004 Oct 15;279(42):44101-12. Epub 2004 Aug 5.
• Citations: 2 Articles

Backbone

• Vector backbone: pcDNA1/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 7103
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: G-alpha-s-EE-ECFP
• Alt name: Gnas
• Species: R. norvegicus (rat)
• Insert Size (bp): 2303
• Mutation: His was substituted for Asn164 in ECFP (Clontech). BamHI sites in the alpha-s cDNA were removed and alpha-s residues 73-84 were replaced with Gly-Ser adding a unique BamHI site into which ECFP was subcloned.
• Entrez Gene: Gnas (a.k.a. ALEX, G-alpha-8, Gnas1, Gnpas, Nesp55, SCG6)
• Promoter: CMV
• Tags / Fusion Proteins:
  • ECFP flanked by SGGGS on each side was inserted in place of alpha-s residues 73-84.
  • internal EE epitope (residues 189-194 in alpha-s sequence are EYMPTE)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Alpha-s-EE was produced in the laboratory of Henry Bourne. ECFP purchased from Clontech.

Terms and Licenses

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

Depositor Comments

Reference for alpha-s-EE: Wilson, P. T., and Bourne, H. R. (1995) Fatty acylation of αz. Effects of palmitoylation and myristoylation on αz signaling. J. Biol. Chem. 270, 9667-9675

DNA (Catalog # 55793-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 # 55793-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.