pETDuet S2
(Plasmid #171921)

• Purpose: Express RNA-based (p)ppGpp sensor and a NirFP
• Depositing Lab: Mingxu You
• Publication: Sun et al Angew Chem Int Ed Engl. 2021 Nov 2;60(45):24070-24074. doi: 10.1002/anie.202111170. Epub 2021 Oct 5.

Backbone

• Vector backbone: pETDuet-1
• Backbone manufacturer: EMD Biosciences
• Backbone size w/o insert (bp): 5267
• Total vector size (bp): 6108
• Vector type: Bacterial 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 1

• Gene/Insert name: RNA-based (p)ppGpp sensor
• Species: Synthetic
• Insert Size (bp): 136
• Promoter: T7

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SgrAI (not destroyed)
• 3′ cloning site: SacII (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7term

Gene/Insert 2

• Gene/Insert name: NirFP
• Alt name: eqFP670
• Species: Synthetic
• Insert Size (bp): 705
• Promoter: T7

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7term

Resource Information

• Supplemental Documents:
  • pETDuet Broccoli-based ppGpp Sensor (S2).gb
• A portion of this plasmid was derived from a plasmid made by: The pETDuet vector with eqFP670 cloned was received from Samie R. Jaffrey lab at Weill Medical College, Cornell University.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Addgene QC identified a C66Y change in the lacI coding sequence that may impact the leakiness of expression of the (p)ppGpp sensor and NirFP insert.