pORTMAGE-3
(Plasmid #72678)

• Purpose: Expresses Lambda Red recombinases and a dominant negative MutL allele all controlled by temperature sensitve cI857 repressor for high precision and efficiency MAGE experiments. Kan resistance marker.
• Depositing Lab: Csaba Pál
• Publication: Nyerges et al Proc Natl Acad Sci U S A. 2016 Feb 16. pii: 201520040.

Backbone

• Vector backbone: pSIM8
• Backbone manufacturer: Datta S. et al. (Gene 379 (2006) 109-115)
• Backbone size w/o insert (bp): 6248
• Total vector size (bp): 8096
• Modifications to backbone: MutL E32K allele cloned into vector
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Induction of expression of Lambda recombinases and MutL E32K allele at 42°C for 15 minutes, otherwise grow at 30°C
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mutL E32K
• Species: Escherichia coli
• Insert Size (bp): 1848
• Mutation: E32K mutation conferring dominant mutator phenotype
• Tag / Fusion Protein:
  • none (N terminal on insert)

Resource Information

• Supplemental Documents:
  • pORTMAGE flow chart
  • pORTMAGE & pSEVAMAGE Protocol
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

Please visit the depositor's website: http://group.szbk.u-szeged.hu/sysbiol/pal-csaba-lab-resources.html for additional resource information.