pLT06
(Plasmid #255019)

• Purpose: (Empty Backbone) Markerless deletion vector that enables manipulation of enterococcus to either insert DNA or delete DNA from the genome. Contains resistance gene for chloramphenicol resistance.
• Depositing Lab: Lynn Hancock
• Publication: Thurlow et al J Bacteriol. 2009 Oct;191(20):6203-10. doi: 10.1128/JB.00592-09. Epub 2009 Aug 14.

Backbone

• Vector backbone: pcjk47, pgb354, pks05
• Selectable markers: pheS counterselectable marker using the compound p-Cl-phenylalanine

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol (10 μg/mL) and Kanamycin (50 μg/mL)
• Growth Temperature: 30°C
• Growth Strain(s): EC1000 (Addgene #71852)
• Growth instructions: Continually maintain growth culture below 30 °C. While conventional E. coli cloning hosts will work, you may have more success using strain EC1000 which has an integrated copy of a functional repA in the genome. Strain EC1000 should be grown with kanamycin at 50 μg/mL to maintain the chromosomal copy of repA. Any concentration above 10 μg/mL of chloramphenicol will result in plasmid instability, as we have observed gene duplication events with the chloramphenicol acetyl transferase gene on the plasmid.
• Copy number: Low Copy

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This plasmid was built by combining components of plasmids pCJK47, chloramphenicol resistance gene from pGB354, and the temperature-sensitive replicon from plasmid pCASPER.

Terms and Licenses

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