Lu SPECS Library
(Pooled Library #127842)

• Purpose:

  The Synthetic Promoters with Enhanced Cell-state Specificity (SPECS) library enables high-throughput screening and promoter identification.
  
  Library constructs are composed of tandem repeats of transcription factor binding sites upstream of an adenovirus minimal promoter controlling expression of mKate2.

• Vector Backbone:

  FUW (Plasmid #14882)

• Depositing Labs: Timothy Lu
• Publication: Wu et al Nat Commun. 2019 Jun 28;10(1):2880. doi: 10.1038/s41467-019-10912-8.

Library Details

• Species: Human
• Synthetic promoters: 6,107 unique motifs
• Lentiviral Generation: 3rd

Library Shipping

This library is delivered as suspended DNA 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: 50 ng/µL

Resource Information

• Protocols:
  • Library Amplification, Screening and NGS protocols (DOCX, 19.3 KB)
• Depositor Data:
  • Primers, Promoter Sequences, NGS Schema and Library Coverage (XLSX, 905.8 KB)
• Scripts:
  • The depositing lab has shared their analysis pipeline in their GitHub repository.
• Articles Citing this Pooled Library:
  • SPECS (#127842)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA
• Evrogen Limited Use Label License for FPs

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

Figure 1: Overview of Lu Lab SPECS Library construction and use. Image adapted from Wu et al. 2019 under CC-BY 4.0 International License .

The Lu Lab has provided two accessory plasmids to accompany the SPECS Library:

• Plasmid #128569: Accessory plasmid 1 - Mix with the library during lentiviral production
• Plasmid #128571: Accessory plasmid 2 - Used to clone identified synthetic promoters

NOTE: Lentiviral vectors can recombine during library amplification, resulting in an ~1–1.7 kb band containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the PuroR gene and will not be selected during screening. If this ~1.7 kb band is observed after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.