pET29b-H6_Streptavidin_sfGFP
(Plasmid #124296)

• Purpose: His6-tagged Streptavidin GFP fusion protein
• Depositing Lab: Mark Arbing
• Publication: Streptavidin-GFP fusion (pET29b) (unpublished)

Backbone

• Vector backbone: pET29b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5237
• Total vector size (bp): 6500
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: His6 tag - TEV protease site - T7 tag - Streptavidin (15-159) - superfolder GFP
• Species: Synthetic
• Insert Size (bp): 1263
• Tag / Fusion Protein:
  • His6 and T7 tags (N terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • pet29b-streptavidin_sfgfp.gb

Terms and Licenses

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

Depositor Comments

T7 promoter so expression requires DE3 strain. Pilot experiments using E. coli BL21(DE3) indicate 18C expression overnight gives higher yields than 37C (for 4 hours). The fusion protein was soluble and purified by Ni-NTA chromatography using 50 mM Tris pH 8.0, 300 mM NaCl, 10/300 mM imidazole. Size exclusion chromatography indicated that the fusion protein was tetrameric in Tris-buffered saline; no aggregates were present.