Cer(1-158)-gamma-5 in pcDNAI/Amp
(Plasmid #55625)

• Purpose: An amino-terminal mCerulean fragment was fused to Ggamma-5. When co-expressed with a carboxyl-terminal CFP fragment fused to a Gbeta subunit with which it interacts, a fluorescent signal is produced.
• Depositing Lab: Catherine Berlot
• Depositing Organization: Geisinger Health Center
  Located in United States of America
• Publication: Mervine et al Mol Pharmacol. 2006 Jul;70(1):194-205. Epub 2006 Apr 26.

Backbone

• Vector backbone: pcDNAI/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 5458
• 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(1-158)-gamma-5
• Alt name: GNG5
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 658
• Mutation: Ggamma-5 was amplified by PCR, which added a BamHI site and a linker sequence (Arg-Ser) to the 5' end and a 3' BglII site. Cloning into the BglII site of Cer(1-158) destroyed the BamHI site.
• GenBank ID: NP_005265.1
• Entrez Gene: GNG5
• Promoter: CMV
• Tag / Fusion Protein:
  • Cer(1-158) was fused to the amino terminus of Ggamma-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

• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: Monomeric Cerulean was obtained from David Piston, Vanderbilt University, Nashville, TN. Gamma-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.

DNA (Catalog # 55625-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 # 55625-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.