CAG::PSAML141F,Y115F:GlyR-IRES-GFP
(Plasmid #32480)

• Depositing Lab: Scott Sternson
• Publication: Magnus et al Science. 2011 Sep 2;333(6047):1292-6.

Backbone

• Vector backbone: pCAGGS- IRES-GFP
• Backbone size w/o insert (bp): 7200
• Modifications to backbone: WPRE (Woodchuck hepatitis virus posttranscriptional regulatory element) before polyA sequence
• 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: PSAML141F,Y115F:GlyR
• Species: H. sapiens (human)
• Insert Size (bp): 1295
• Promoter: CAG
• Tag / Fusion Protein:
  • IRES-GFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CTGCTAACCATGTTCATGC
• 3′ sequencing primer: GAAGACTCTTGCGTTTCTG

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Note on the difference between PSAMY115F,L141F:GlyR and PSAML141F:GlyR silencers:
In Magnus et al, the PSAMY115F,L141F-GlyR channel was emphasized for silencing because this channel has the lowest acetylcholine responsiveness. However, PSAML141F-GlyR constructs, lacking the Y115F mutation, already have low ACh potency. The Y115F mutation also slightly increases the EC50 for the activator ligand, PSEM89S. Since PSAML141F-GlyR constructs already have very low acetylcholine potency, in many cases it may not be worth the slight reduction in PSEM potency. We have made both constructs available for researchers who may find that they have different requirements with respect to acetylcholine responsiveness.