mCherryS131C
(Plasmid #48732)

• Purpose: Expresses mCherryS131C in E. coli cells
• Depositing Lab: Bradley Olsen
• Publication: Thomas et al ACS Nano. 2011 Jul 26;5(7):5697-707. doi: 10.1021/nn2013673. Epub 2011 Jun 22.

Backbone

• Vector backbone: pQE-9
• Backbone manufacturer: QIAGEN
• Backbone size w/o insert (bp): 3439
• Total vector size (bp): 4136
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): SG13009 (QIAGEN)
• Growth instructions: The SG13009 strain contains the repressor plasmid pREP4 to constitutively express the lac repressor and enable trans repression of protein expression before IPTG induction. The pREP4 plasmid is maintained by kanamycin selection.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: mCherryS131C
• Alt name: mCherry
• Species: Synthetic
• Insert Size (bp): 861
• Mutation: changed serine 131 to cysteine
• Promoter: T5
• Tag / Fusion Protein:
  • 6xHis (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: HindIII (unknown if destroyed)
• 5′ sequencing primer: CCCGAAAAGTGCCACCTG
• 3′ sequencing primer: CAT-R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid must be grown under Ampicillin (100ug/ml) and Kanamycin (50ug/ml) selection in order to retain both the mCherryS131C plasmid and the pREP4 helper plasmid.

Please note that verifying this plasmid by sequencing or digest is challenging due to the presence of the helper plasmid, as multiple bands or mispriming is likely to occur. See Addgene's sequencing results for recommended primers.

The S131C mutation in this plasmid is S136C, when compared to the starting Met of wild-type mCherry.