pET28-mEGFP(A206K)-TwStrep
(Plasmid #208622)

• Purpose: for E. coli expression of C-ter Twin Strep tagged mEGFP(A206K), EGFP cloned from pEGFP-C1, A206K introduced by seamless (Gibson) clone
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pET28-3'TwStrep
• Modifications to backbone: refer to (Addgene#208053)
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: we use BL21 (DE3) for expression
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: mEGFP(A206K)
• Alt name: mEGFP
• Alt name: GFP
• Alt name: monomer enhanced green fluoresence protein
• Species: Synthetic; Aequorea victoria
• Mutation: (compared with pEGFP-C1, protein level) A206K
• GenBank ID: U55763 AAB02576
• Promoter: T7
• Tags / Fusion Proteins:
  • Twin Strep (C terminal on backbone)
  • 6xHis (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: pETupstream, T7p
• 3′ sequencing primer: T7ter

Resource Information

• Supplemental Documents:
  • pET28-mEGFP(A206K)-TwStrep.gb
• A portion of this plasmid was derived from a plasmid made by: EGFP sequence clone from comercial plasmid pEGFP-C1 (TAKARA-Clontech), A206K mutation was adopted from Benjamin Glick's work (Addgene#36412)

Terms and Licenses

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

Depositor Comments

>Quick Charge Primers for EGFP A206K
EGFP-A206K-F: 5' CCTGAGCACCCAGTCCAAGCTGAGCAAAGACCCCA 3'
EGFP-A206K-R: 5' TGGGGTCTTTGCTCAGCTTGGACTGGGTGCTCAGG 3'