pH6-2xmNeonC
(Plasmid #178463)

• Purpose: A tandem mNeonGreen fluorescent protein (2xmNeon) with an N-terminal 6xHis-tag and C-terminal cysteine in pET28a.
• Depositing Lab: Siegfried Musser
• Publication: Sharma et al Commun Biol. 2022 Sep 19;5(1):988. doi: 10.1038/s42003-022-03952-2.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5303
• Total vector size (bp): 6755
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: mNeonGreen dimer
• Species: Branchiostoma lanceolatum
• Insert Size (bp): 1476
• Mutation: Free cysteine at the C-terminus of the dimer
• GenBank ID: KC295282.1
• Promoter: T7
• Tag / Fusion Protein:
  • 6xHis (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: taatacgactcactatagg
• 3′ sequencing primer: ggccccaaggggttatgc

Resource Information

• Supplemental Documents:
  • pH6-2xmNeonC.gb
• A portion of this plasmid was derived from a plasmid made by: Addgene #69146

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • mNeonGreen Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The mNeonGreen coding sequence was PCR amplified from pH6-mNeon (Addgene #178460) with an SpeI site at both ends and inserted into the vector fragment of pH6-mNeonC (Addgene #178461) digested with SpeI.