pET28-3'TwStrep
(Plasmid #208053)

• Purpose: (Empty Backbone) Empty vector for prokaryotic expression, insert a Twin Strep tag (with TAA) between HindII and XhoI site of pET28a (+), both HindIII and XhoI are retained. HindIII in-frame with Twin Strep tag.
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pET28a(+)
• Backbone manufacturer: Merck-Millipore (Novagen)
• Modifications to backbone: add Twin Strep tag
• Vector type: Bacterial Expression
• Promoter: T7
• Tag / Fusion Protein:
  • Twin Strep (C terminal on backbone)

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

Resource Information

• Supplemental Documents:
  • pET28-3'TwStrp.gb
• A portion of this plasmid was derived from a plasmid made by: Strep tag is ® of IBA lifescience.

Terms and Licenses

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

Depositor Comments

We optimized nucleic acid sequence of Twin Strep, eliminated tandem repeats, for better oligo assembly. The nucleic acids are different from that of IBA lifescience, amino acids are the same. The Twin Strep tag was synthesized by oligo assembly using following oligos:
# Sequence 5'-3' bp
pET28-TwStrp.o1: CGACAAGCTTTCCGGAGGGAGCGCTTGGAGCCACCCGCAGTTCGAA 46
pET28-TwStrp.o2: CGATCCACCGCCAGAACCTCCACCTTTTTCGAACTGCGGGTGGCTC 46
TwStrp.opt.o3: AGGTTCTGGCGGTGGATCGGGAGGTTCAGCGTGGTCTCACCCCCAA 46
TwStrp.opt.o4: GGTGCTCGAGCTATTACTTCTCAAATTGGGGGTGAGACCACGCT 44