CHRM5-Tango
(Plasmid #66252)

• Purpose: Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation
• Depositing Lab: Bryan Roth
• Publication: Kroeze et al Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20.

Backbone

• Vector backbone: empty Tango
• Backbone size w/o insert (bp): 6632
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: CHRM5
• Species: H. sapiens (human)
• Insert Size (bp): 1596
• Entrez Gene: CHRM5 (a.k.a. HM5)
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: Tet-R

Terms and Licenses

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

Depositor Comments

Please note: Most of the PRESTO-Tango vectors do not contain the XhoI cut site downstream of the tTA sequence and instead carry an XbaI site. If you need to perform an XhoI digest we recommend sequencing with a forward primer at the C-terminus of tTA to determine which of the two sites is present. Suggested primer: 5-gagctccacttagacggcgagg-3. Original backbone was pcDNA3.1(+). These plasmids were generated as part of the Illuminating the Druggable Genome (IDG) program sponsored by the NIH Common Fund. The goal of this program is to identify, gather, and distribute information and resources for proteins that currently are not well-studied yet belong to commonly drug-targeted protein families: protein kinases, non-olfactory G-protein coupled receptors (GPCRs), and ion channels. The IDG program is designed to develop fundamental research tools for understudied proteins, elucidate their function, and disseminate the IDG-related resources and data to the greater scientific community.