pCURE
(Plasmid #23013)

• Depositing Lab: James Bowie
• Publication: Massey-Gendel et al Protein Sci. 2009 Feb . 18(2):372-83.

Backbone

• Vector backbone: pSC101ts
• Backbone manufacturer: N/A
• Backbone size w/o insert (bp): 8900
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Tetracycline, 10 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Top10/P3
• Growth instructions: Propagate at 30 degrees Celsius and 5ug/mL tetracylcine.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: I-CreI homing endonuclease
• Species: Chlamydomonas reinhardtii
• Insert Size (bp): 2000
• Mutation: I-CreI expression is controlled by the arabinose promoter.
• GenBank ID: P05725

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: PvuII (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a

Resource Information

• A portion of this plasmid was derived from a plasmid made by: I derived code for I-CreI behind the arabinose promoter and the arabinose regulatory elements from plasmid pAE from Lenny Seligman of Pomona College, an author on the cited paper describing the plasmid.

Terms and Licenses

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

Depositor Comments

Seligman, LM et al., 1997, Genetics, 147: 1653-1664

The plasmid can be propagated in 0.2% glucose to keep expression levels low and to guard against mutations.

Addgene's quality control sequence shows several sequence discrepancies with the available NCBI reference sequence for AraC and ICreI. Depositor tested this plasmid and found the plasmid can be used to cure cells of other plasmids that contain an I-CreI cut site, and the pCURE can be removed from cells via its temperature sensitive origin of replication. Hence, these mutations are not detrimental to the plasmid's function.