Cer(1-158)-beta-1 in pcDNAI/Amp
(Plasmid #55707)

• Purpose: An amino-terminal fragment of mCerulean was fused to Gbeta1. When co-expressed with a carboxyl terminal CFP fragment fused to a Ggamma subunit, a fluorescent signal s produced.
• Depositing Lab: Catherine Berlot
• Publication: Yost et al Mol Pharmacol. 2007 Oct;72(4):812-25. Epub 2007 Jun 27.

Backbone

• Vector backbone: pcDNA1/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6283
• Modifications to backbone: BglII introduced to polylinker of pcDNA1/Amp 3' of 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: mCer(1-158)-beta 1
• Alt name: transducin beta
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1483
• Mutation: GBeta-1 was amplified via PCR, which removed an internal BglII site and 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: GNB1 (a.k.a. HG2A, MDS, MRD42)
• Promoter: CMV
• Tag / Fusion Protein:
  • Cer(1-158) was fused to the amino terminus of Gbeta1. (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 was obtained from David Piston, Vanderbilt University, Nashville, TN. Beta-1 cDNA obtained from Janet Robishaw, Weis Center for Research, Geisinger Clinic, Danville PA
• Article Citing this Plasmid:
  • 1 Reference

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.