pET14_GFP5_Encap
(Plasmid #86408)

• Purpose: Expresses sfGFP tagged with a 5 amino acid CLP and encapsulin as an operon
• Depositing Lab: David Savage
• Publication: Cassidy-Amstutz et al Biochemistry. 2016 Jun 21;55(24):3461-8. doi: 10.1021/acs.biochem.6b00294. Epub 2016 Jun 7.

Backbone

• Vector backbone: pET14b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 6600
• Modifications to backbone: BsaI and BsmBI recognition sites were removed
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: sfGFP with a 5aa CLP tag
• Species: Thermatoga maritima
• Insert Size (bp): 800
• Promoter: T7

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: BsaI (destroyed during cloning)
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: T7 reverse

Gene/Insert 2

• Gene/Insert name: Encapsulin
• Species: Thermatoga maritima
• Insert Size (bp): 800
• Promoter: T7
• Tag / Fusion Protein:
  • 6x Histidine (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: BsaI (destroyed during cloning)
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: T7 reverse

Resource Information

• Supplemental Documents:
  • pDQ03 sfGFP_5aaFLP + opt. Encapsulin in pET14GG CHis.gb

Terms and Licenses

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