hGFAP-fLuc
(Plasmid #40589)

• Depositing Lab: Albee Messing
• Depositing Organization: University of Wisconsin, Madison
  Located in United States of America
• Publication: Cho et al J Neurochem. 2009 Jul;110(1):343-51. Epub 2009 May 5.
• Citations: 3 Articles

Backbone

• Vector backbone: pGL3-basic
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 4818
• Vector type: Luciferase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: hGFAP promoter
• Alt name: hGFAP
• Alt name: GFAP promoter
• Alt name: GFAP
• Species: H. sapiens (human)
• Insert Size (bp): 2200
• Mutation: Contains human GFAP promoter fragment (-2163 to +47). ATG codon at position +15 changed to TTG (transcription starts at the firefly luciferase ATG at position +102)
• Entrez Gene: GFAP (a.k.a. ALXDRD)
• Promoter: GFAP promoter
• Tag / Fusion Protein:
  • Firefly luciferase (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: BglII (destroyed during cloning)
• 5′ sequencing primer: RVprimer3
• 3′ sequencing primer: LucNrev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The 2.2 kb human GFAP promoter fragment was derived from the GFAP-lacZ transgene previously described by Brenner et al. (1994).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

To construct a transgene expressing firefly luciferase regulated by the GFAP promoter (GFAP-fLuc), the depositor utilized the 2.2 kb human GFAP promoter derived from the GFAP-lacZ transgene previously described by Brenner et al. (1994). The GFAP promoter fragment (-2163 to +47) was subcloned into the Bgl II site in the pGL-3 basic vector (Promega, Madison, WI, USA) containing the firefly luciferase gene. The transgene was sequenced to confirm correct orientation and sequence.

DNA (Catalog # 40589-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
  • Luciferase Limited Use Label License
• 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 # 40589-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
  • Luciferase Limited Use Label License
• 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.