YFP(1-158)-beta-5 in pcDNAI/Amp
(Plasmid #54470)

• Purpose: An amino-terminal YFP fragment was fused to Gbeta-5. When co-expressed with a carboxyl-terminal YFP or CFP fragment fused to a Ggamma subunit, a fluorescent signal is produced.
• Depositing Lab: Catherine Berlot
• Depositing Organization: Geisinger Health Center
  Located in United States of America
• Publication: Hynes et al J Biol Chem. 2004 Jul 16;279(29):30279-86. Epub 2004 May 10.
• Citations: 2 Articles

Backbone

• Vector backbone: pcDNA1/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6322
• Modifications to backbone: A BglII site was introduced into 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: YFP(1-158)/beta-5
• Alt name: gnb5
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1522
• Mutation: YFP (1-158) includes a substitution of Met for Gln-69. 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 YFP(1-158) destroyed the BamHI site.
• Entrez Gene: GNB5 (a.k.a. GB5, HG2E, IDDCA, LADCI, LDMLS1, LDMLS2, gbeta5)
• Tag / Fusion Protein:
  • YFP(1-158) was fused to the amino terminus of Gbeta-5. (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: pEYFP purchased from Clontech. 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

DNA (Catalog # 54470-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 # 54470-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.