pAAV-CAG-H2BmGreenLantern
(Plasmid #177332)

• Purpose: To express a bright monomeric green FP to label eucaryotic cell nuclei. To be used in clearing and other fluorescent microscope methods.
• Depositing Labs: Murray Blackmore, Gregory Petsko, Pantelis Tsoulfas
• Publication: Wang et al Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.

Backbone

• Vector backbone: pAAV-CAG-GFP
• Backbone manufacturer: Boyden laboratory
• Backbone size w/o insert (bp): 4714
• Total vector size (bp): 5858
• Modifications to backbone: The GFP of the original vector was excised with XbaI and XhoI and replaced with the H2BmGL
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: H2B
• Species: R. norvegicus (rat); Rat
• Insert Size (bp): 1144
• Mutation: This fusion protein was optimized to the Human codon usage.
• GenBank ID: NP_001100822
• Entrez Gene: Hist1h2bq (a.k.a. Hist1h2bh)
• Promoter: CAG
• Tag / Fusion Protein:
  • mGreenLantern (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI or HindIII (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: pCAG-F or pCAGGS-5
• 3′ sequencing primer: WPRE-R

Resource Information

• Supplemental Documents:
  • pAAV-CAG-H2BmGreenLantern

Terms and Licenses

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

Depositor Comments

For more information, also see Campbell et al. 2020 "mGreenLantern: a bright monomeric fluorescent protein with rapid expression and cell filling properties for neuronal imaging" PNAS, https://doi.org/10.1073/pnas.2000942117.