pKM461
(Plasmid #108320)

• Purpose: For performing ORBIT - Expresses Che9c phage RecT and Bxb1 phage Integrase, and contains SacB
• Depositing Lab: Kenan Murphy
• Publication: Murphy et al MBio. 2018 Dec 11;9(6). pii: mBio.01467-18. doi: 10.1128/mBio.01467-18.

Backbone

• Vector backbone: pUV15tet-O
• Backbone manufacturer: Dirk Schnappinger and Sabine Ehrt
• Modifications to backbone: Insertion of RecT, Bxb1 Integrase, and SacB. Removal of GFP and Hyg.
• 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: Che9C RecT
• Species: mycobacterial phage
• Insert Size (bp): 1062

Gene/Insert 2

• Gene/Insert name: Bxb1 Integrase
• Species: mycobacterial phage
• Insert Size (bp): 1503

Gene/Insert 3

• Gene/Insert name: Tet Repressor
• Species: bacterial
• Insert Size (bp): 624

Gene/Insert 4

• Gene/Insert name: SacRB
• Species: B. subtilis
• Insert Size (bp): 1785

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Julia van Kessel and Graham Hatfull (RecT generated from pJV53); Michele Calos (Addgene plasmid pCMV-Bx was the source for Bxb1 Integrase; Adrie J. C. Steyn supplied pCre-SacB-kan, the source for SacB.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

The plasmid is induced with 500 ng/ml anhydrotetracycline to induce expression of RecT and Bxb1 Integrase for performing the ORTBIT protocol.