pIMAY*
(Plasmid #121441)

• Purpose: (Empty Backbone) Allelic exchange plasmid with PheS* counter-selection for Staphylococcus aureus
• Depositing Lab: Angelika Grundling
• Publication: Schuster et al Microbiology. 2019 Apr 3. doi: 10.1099/mic.0.000791.

Backbone

• Vector backbone: pIMAY*
• Backbone size (bp): 5560
• Vector type: Escherichia coli/Staphylococcus aureus shuttle vector
• Selectable markers: Chloramphenicol (S. aureus)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: This is an Escherichia coli/Staphylococcus aureus shuttle vector. For S. aureus, grow at 28°C with 10 µg/ml Chloramphenicol.
• Copy number: Low Copy

Cloning Information

• Cloning method: Restriction Enzyme

Resource Information

• Supplemental Documents:
  • pIMAY_STAR_final.gb
  • piMAY Knockout Protocol
• A portion of this plasmid was derived from a plasmid made by: The Tim Foster lab, plasmid pIMAY* originated from pIMAY (https://www.addgene.org/68939/ ). Nucleic acid for counter-selector pheS* was synthesized de novo.
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

Always check if the pIMAY* plasmid is still temperature sensitive in Staphylococcus aureus before attempting allelic exchange (see accompanying protocol).

This plasmid has been found to be somewhat unstable and prone to concatenation. Concatenation often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.