pET21c-His-Xa-Gluc-Xa-C9D-TGinsert
(Plasmid #124660)

• Purpose: reporter protein which emits bright blue fluorescence
• Depositing Lab: Yutaka Kuroda
• Publication: Wu et al Sci Rep. 2020 Nov 18;10(1):20069. doi: 10.1038/s41598-020-76486-4.

Backbone

• Vector backbone: pET21c
• Backbone size w/o insert (bp): 5562
• Total vector size (bp): 6158
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Gaussia luciferase-C9D-TGinsert
• Alt name: Gluc-C9D-TGinsert
• Species: Gaussia princeps
• Insert Size (bp): 600
• Mutation: changed glutamic acid 100 to alanine, and glycine 103 to arginine
• GenBank ID: AY015993.1 AY015993.1
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • MHHHHHHIEGRTG（His-Tag,FactorXa,TG） (N terminal on insert)
  • IEGRTMGDDDGDDDGDDD（FactorXa,TMG,C9D-Tag） (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeⅠ (unknown if destroyed)
• 3′ cloning site: BamHⅠ (unknown if destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminater

Terms and Licenses

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

Depositor Comments

This plasmid is described in the following publications:
1. Wu nan, Tomonori Saotome, Satoru Unzai, Yutaka Kuroda* and Toshio Yamazaki*, Solution structure of Gaussia Luciferase with five disulfide bonds and identification of a putative coelenterazine binding cavity by heteronuclear NMR, Scientific Reports volume 10, Article number: 20069 (2020)
2. Tharangani Rathnayaka, Minako Tawa, Takashi Nakamura, Shihori Sohya, Kunihiro Kuwajima, Masafumi Yohda , Yutaka Kuroda*, Solubilization and folding of a fully active recombinant Gaussia luciferase with native disulfide bonds by using a SEP-Tag, Biochim Biophys Acta. Proteins and Proteomics, 1814(12):1775-1778(2011)