pGFAP::MyrGFP
(Plasmid #22672)

• Purpose: This plasmid has been discontinued and a new version is now available. Please see: pGFAP::MyrGFP_v2 (Plasmid #160277) https://www.addgene.org/160277/
• Depositing Lab: Robert Benezra
• Publication: Nam et al Cell Stem Cell. 2009 Nov 6. 5(5):515-26.

Backbone

• Vector backbone: Gfa2-cLac
• Backbone size w/o insert (bp): 5500
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: myrGFP
• Insert Size (bp): 700
• Mutation: Gfa2 human GFAP promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • pGfa2-myrEGFP corrected.gb
• A portion of this plasmid was derived from a plasmid made by: Michael Brenner [mailto:]
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

MyrEGFP fragment amplified by high fidelity PCR from plasmid pROSA26-mT-mG (www.addgene.org/17787/) was InFusion cloned into BamHI site of the pGFAP-EGFR* plasmid obtained from Dr. Eric Holland. The ~2.2 kb GFAP promoter in this plasmid is presumably the same as the one utilized in the GFAP-tva transgenic mouse in Holland EC and Varmus HE https://doi.org/10.1073/pnas.95.3.1218. Addgene NGS sequencing confirmed the Gfa2 promoter sequence in this plasmid is identical to the hGFAP promoter sequences in other Gfa2 plasmids, such as hGFAP-GFP (http://www.addgene.org/40592/) and hGFAP-Cre (http://www.addgene.org/40591/). One base pair is missing immediately upstream of the stop codon. The myrEGFP sequence is not perturbed except the very last amino acid. However, the mouse Protamine 1 sequence that is downstream of the myrEGFP sequence and is otherwise silent is now translated. Thus, a fusion protein of membrane-anchored myrEGFP and mPrm1 is translated.