CFP(1-158)-RGS7t
(Plasmid #55761)

• Purpose: An amino-terminal CFP Fragment was fused to residues 202-477 of RGS7. When co-expressed with a carboxyl terminal fragment of CFP fused to Gbeta-5, a fluorescent signal is produced.
• Depositing Lab: Catherine Berlot
• Publication: Yost et al Mol Pharmacol. 2007 Oct;72(4):812-25. Epub 2007 Jun 27.

Backbone

• Vector backbone: pcDNAI/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6091
• Modifications to backbone: A BglII site was introduced into the polylinker of pcDNA1/Amp 3' to the BamHI site.
• 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: RGS7(202-477)-CFP(1-158)
• Alt name: RGS7
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1291
• Mutation: The RGS sequence amino terminal to the GGL domain was deleted by amplifying only residues 202-477 in a PCR reaction that added a BamHI site and a linker sequence (RSIAT) to the 5' end and a 3' BglII site. Cloning into the BglII site of CFP(1-158) destroyed the BamHI site.
• GenBank ID: AF090116.1
• Entrez Gene: RGS7
• Promoter: CMV
• Tag / Fusion Protein:
  • CFP(1-158) was fused to the amino terminus of RGS7(202-477). (N terminal on insert)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: RGS7-S1 cDNA was purchased from Guthrie cDNA Resource Center, Sayre, PA (now Missouri S&T cDNA Resource Center, Rolla, MO). pECFP purchased from Clontech.

Terms and Licenses

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