Cer(159-238)-beta-5 in pcDNAI/Amp
(Plasmid #55778)

• Purpose: A carboxyl-terminal mCerulean fragment was fused to Gbeta-5. When co-expressed with an amino terminal mCerulean fragment fused to a Ggamma subunit, 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): 6097
• Modifications to backbone: A BglII site was introduced into the polylinker of pcDNAI/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: Cer(159-238)-beta-5
• Alt name: gnb5
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1297
• Mutation: GBeta-5 was amplified via PCR, which added an N-terminal linker sequence (RSIAT), a 5' BamHI site, and a 3' BglII site. Cloning into the BglII site of Cer(1-158) destroyed the BamHI site.
• Entrez Gene: GNB5 (a.k.a. GB5, HG2E, IDDCA, LADCI, LDMLS1, LDMLS2, gbeta5)
• Promoter: CMV
• Tag / Fusion Protein:
  • mCer(159-238) was fused to the amino terminus of Gbeta5. (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: Monomeric Cerulean obtained from David Piston, Vanderbilt University, Nashville, TN. Beta-5 cDNA was obtained from Janet Robishaw, Weis Center for Research, Geisinger Clinic, Danville, PA.

Terms and Licenses

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

Depositor Comments

Reference for monomeric Cerulean:

Rizzo MA, Springer GH, Granada B, Piston DW. An improved cyan fluorescent protein variant useful for FRET. Nat Biotechnol. 2004;22(4):445-9. PubMed PMID: 14990965.