pOPINE GFP nanobody:Halo:His6
(Plasmid #111090)

• Purpose: Bacterial expression of a fusion protein consisting of a GFP nanobody (Dr. Brett Collins, Addgene plasmid # 49172), the Halo tag and a His6 tag.
• Depositing Lab: Lennart Wirthmueller
• Publication: Chen et al Front Plant Sci. 2018 Nov 1;9:1581. doi: 10.3389/fpls.2018.01581. eCollection 2018.

Backbone

• Vector backbone: pOPINE
• Backbone manufacturer: Berrow et al Nucleic Acids Res. 2007 . 35(6):e45.
• Backbone size w/o insert (bp): 5163
• Total vector size (bp): 6417
• Modifications to backbone: none
• Vector type: Mammalian Expression, Bacterial Expression, Insect 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

• Gene/Insert name: GFP Nanobody
• Alt name: abGFP4 PMID 16095608
• Alt name: Kubala et al Protein Sci. 2010 Dec;19(12):2389-401. doi: 10.1002/pro.519.
• Species: Synthetic
• Insert Size (bp): 1254
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • Halo (C terminal on insert)
  • His6 (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: GFP Nanobody; Addgene plasmid #49172 (Dr. Brett Collins) Halo tag; Promega
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • HaloTag Vector Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Dr. Brett Collins has given his consent to making the modified GFP nanobody plasmid available at Addgene. The Halo tag can be distributed under the HaloTag Vector Limited Use Label License (Promega).

Please visit https://figshare.com/articles/Production_of_magnetic_GFP_affinity_beads/6894248 a protocol describing the production of GFP affinity beads using this plasmid.

Please visit https://www.biorxiv.org/content/early/2018/04/20/304527 for bioRxiv preprint.