pHL 100
(Plasmid #52989)

• Purpose: pLlac0-1:OxyS-mCherry, pLtetO-1:fhlA::gfp
• Depositing Lab: Han Lim
• Publication: Hussein et al Proc Natl Acad Sci U S A. 2011 Jan 18;108(3):1110-5. doi: 10.1073/pnas.1010082108. Epub 2010 Dec 28.

Backbone

• Vector backbone: unknown
• Backbone manufacturer: Lim lab
• Vector type: Bacterial Expression, Synthetic Biology

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: OxyS sRNA
• Species: E.coli
• Promoter: pLlac0-1
• Tag / Fusion Protein:
  • RBS-mCherry (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: unknown
• 3′ sequencing primer: mCherry-R

Gene/Insert 2

• Gene/Insert name: fhlA
• Alt name: target mRNA
• Species: E. coli
• Mutation: −309 to +96 bp relative to start codon
• Promoter: pLtetO-1
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: unknown
• 3′ sequencing primer: gfp-R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The transcription of each sRNA was controlled by the pLlacO-1 promoter, which was induced by adding isopropyl-β-d-thiogalactopyranoside (IPTG) to the media. The part of the target mRNA that is necessary for sRNA regulation was fused to gfp and constitutively transcribed from the pLtetO-1 promoter without TetR in the system. GFP fluorescence provides a quantitative measure of target gene expression and sRNA activity.