pAG/MNase
(Plasmid #123461)

• Purpose: Expresses Protein A and Protein G IgG binding domains fused to micrococcal nuclease for purposes of CUT&RUN. Includes (his)6 tag for purification and HA-tag for secondary immunoprecipitation.
• Depositing Lab: Steven Henikoff
• Publication: Meers et al Elife. 2019 Jun 24;8. pii: 46314. doi: 10.7554/eLife.46314.

Backbone

• Vector backbone: pK19pA-MN
• Backbone manufacturer: Ulrich Laemmli
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): JM101
• Growth instructions: For plasmid amplification, as well as for expression for purification, it is recommended to use the strain JM101.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Protein A-Protein G-Micrococcal Nuclease
• Alt name: pAG-MNase
• Alt name: pAG/MN
• Alt name: pAG(ERH)_MNase
• Insert Size (bp): 1218
• Promoter: Lac
• Tags / Fusion Proteins:
  • 6xHIS (C terminal on insert)
  • HA (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: M13R
• 3′ sequencing primer: M13F

Resource Information

• Supplemental Documents:
  • Annotated Genbank: pAG_ERH_MNase_6xHIS_HA
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 13 References

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/569129v1 for BioRxiv preprint

This plasmid has been found to be somewhat unstable and prone to multimerization. Multimerization often does not impact plasmid function, but may reduce transformation efficiencies. See our article on Plasmid Dimers and Multimers for more information (https://blog.addgene.org/plasmids-101-dimers-and-multimers).