pNitET-SacB-kan
(Plasmid #107692)

• Purpose: Expression of the phage Che9c RecET system for Recombineering in mycobacteria
• Depositing Lab: Kenan Murphy
• Publication: Murphy et al Methods Mol Biol. 2015;1285:177-99. doi: 10.1007/978-1-4939-2450-9_10.

Backbone

• Vector backbone: pUV15tet-O
• Backbone manufacturer: Sabine Ehrt and Dirk Schnappinger
• Backbone size w/o insert (bp): 8026
• Total vector size (bp): 10820
• Modifications to backbone: RecET inserted in place of GFP; NitR inserted in place of TetR; SacRC inserted.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Che9 phage RecET
• Species: Mycobacteria phage Che9
• Insert Size (bp): 9999

Gene/Insert 2

• Gene/Insert name: PNit2-NitR
• Insert Size (bp): 1075

Gene/Insert 3

• Gene/Insert name: SacRB
• Insert Size (bp): 1785

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Che9 RecET genes were amplified from pJV53, a gift from Graham Hatfull and Julia van Kessel
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This plasmid can be used to promote recombineering in M. smegmatis or M. tuberculosis. The ReET gene are induced with caprolactam in M. smegmatis and with isovaleronitrile (IVN) for M. tuberculosis.

For original description of this methodology, see:
van Kessel, J.C. & Hatfull, G.F. Recombineering in Mycobacterium tuberculosis. Nat Methods 4, 147-152 (2007).