pLY76
(Plasmid #130963)

• Purpose: Engineered sgRNA-LEA2 (2 BoxB aptamers) generator circuit including promoter Plux2
• Depositing Lab: Baojun Wang
• Publication: Liu et al Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.

Backbone

• Vector backbone: p15AC
• Backbone manufacturer: p15A vector from pdCas9-bacteria (Plasmid #44249)
• Backbone size w/o insert (bp): 1725
• Total vector size (bp): 3006
• Modifications to backbone: Adding 2 BbsI sites for construction
• Vector type: Bacterial Expression, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Culture in the Lennox’s Luria-Bertani (LB) medium (10 g/L peptone, 5 g/L yeast extract, 5 g/L NaCl)
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: sgRNA-LEA2
• gRNA/shRNA sequence: AAGTTATTATATAGTTCGGTC
• Species: Synthetic
• Promoter: Plux2

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (destroyed during cloning)
• 3′ cloning site: SpeI (destroyed during cloning)
• 5′ sequencing primer: AACGGTCTGGTTATAGGTACATTGAGCAAC
• 3′ sequencing primer: GTTCGTAAGCCATTTCCGCTCGCCGCAGTC

Terms and Licenses

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

Depositor Comments

The Top10 bacterial strain should be used for downstream applications