6x-His_A1aY1_GFP
(Plasmid #158753)

• Purpose: Bacterial expression of 6xHis-A1aY1 fused with carboxy terminus GFP for recombinant purification
• Depositing Lab: Minhajuddin Sirajuddin
• Publication: Kesarwani et al J Cell Biol. 2020 Oct 5;219(10). pii: 152071. doi: 10.1083/jcb.201912107.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 6067
• Total vector size (bp): 6259
• Modifications to backbone: Inserted A1aY1 sequence in between BamHI and KpnI restriction sites in pET28a vector.
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: 6xHistidine_A1aY1_GFP
• Alt name: 6x His tagged A1aY1-GFP
• Insert Size (bp): 192
• Promoter: T7
• Tags / Fusion Proteins:
  • 6x-Histidine tag (N terminal on insert)
  • Thrombin site (N terminal on insert)
  • T7 leader sequence (N terminal on insert)
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Cloned A1aY1 gene in between BamHI and KpnI sites in pET28a vector having amino-terminus 6x-His sequence and carboxy-terminus GFP sequence.

Terms and Licenses

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