q4wt
(Plasmid #24502)

• Depositing Lab: Bruce Conklin
• Publication: Conklin et al Nature. 1993 May 20. 363(6426):274-6.

Backbone

• Vector backbone: pcDNA1
• Backbone size w/o insert (bp): 4000
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Tetracycline, 100 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10/P3
• Growth instructions: Plasmid contains SupF. Grow in TOP10/P3 cells.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Gq alpha
• Alt name: G protein alpha subunit
• Species: M. musculus (mouse)
• Tag / Fusion Protein:
  • HA (see comments)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: SP6

Resource Information

• Supplemental Documents:
  • G-protein chimera user manual

Terms and Licenses

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

Depositor Comments

This is Gq alpha with an HA epitope engineered into an internal site that does not seem to affect receptor coupling in multiple studies. Epitope tagged by Paul Wilson, see Wedegaertner, JBC 268: 25001-25008. The 5' non-coding sequences were removed, but the 3' non-coding sequences remain, as in Strathmann & Simon (1990) PNAS 87:9113-9117. The parent construct has been donated to the ATCC by the Bourne Lab.

There is a 1bp insertion, a 5bp insertion, and a 1bp deletion when author sequence is aligned with Addgene quality control sequence. These differences are between the stop codon and the SP6 sequence.

Please see supplemental document for G-protein chimera user manual.