attB GPCR Libraries
(Pooled Library #243681, #243682)

• Purpose:

  These libraries encode 767 unique human G protein-coupled receptors (GPCRs) with unique molecular identifiers (UMIs) that can be stably recombined into cell lines modified to genomically encode an attP bxb1 recombination site. Each receptor was encoded using its native codon usage and includes an HA-tag for easy immunostaining, along with an IRES-GFP cassette to monitor transcription and translation fidelity. The platform has been engineered to be compatible with the Golden Gate cloning technology, allowing for the excision of the individual receptors and UMIs from the plasmid backbone and their transfer to any alternative backbone of choice.

  • The attB Barcoded Extended GPCR Library (#243681) contains 767 unique human GPCRs and additionally includes 174 isoforms for selected GPCRs.
  • The attB Barcoded Canonical GPCR Library (#243682) contains 767 unique human GPCRs.
• Vector Backbone:

  pUC19 (Plasmid #50005)

• Depositing Labs: Jonathan Schlebach
• Publication: Tedman et al. bioRxiv 2025.09.19.677468. doi: 10.1101/2025.09.19.677468.

Library Details

• Species: Human
• Insert size: 5–8 kb
• Number of elements:
  
  • Library #243681: 767 unique human GPCRs and 174 isoforms for selected GPCRs.
  • Library #243682: 767 unique human GPCRs.

Library Shipping

Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼20 µL
• Concentration: 100 ng/µL

Resource Information

• Protocols:
  • Library Amplification and Sequencing Preparation Protocols (DOCX, 46 KB)
• Depositor Data:
  • List of Primers (XLSX, 71 KB)
  • Library Index and List of Barcodes (XLSX, 2.1 MB)
  • Library Representation for #243681 (CSV, 28 KB)
• Scripts:
  • Convolution Algorithm (PY, 40 KB)
  • GPCR DRS Dictionary (TSV, 33 KB) (needed to run algorithm)
  • GPCR DRS Configuration (INI, 2 KB) (needed to run algorithm)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

Figure 1: Schematic representation of the region of interest of the plasmids encoded within the GPCR Library. For each plasmid there is an attB site that can recombine with a conjugate attP site which is genomically encoded into a HEK293T cell line engineered by Douglas Fowler (Kenneth et al., 2017; doi: 10.1093/nar/gkx183). Each plasmid also contains a 10-nucleotide unique molecular identifier (UMI) and a corresponding single GPCR with an N-terminal HA Tag. For GPCRs with signal peptides, an HA tag was inserted at a computationally determined suitable location. Downstream of the GPCR there is an Internal Ribosomal Entry Site (IRES) and Dasher-GFP. The GPCR, its UMI and the attB site can be selectively removed using Golden Gate cloning due to the presence of BsmbI sites located immediately before and after the GPCR. Any other BsmbI sites have been computationally and silently removed.