pLJM1-EGFP-GRAM-W
(Plasmid #211704)

• Purpose: Lentiviral expression of EGFP-tagged GRAM domain of GRAMD1b carrying a G187W mutation (pLJM1-EGFP-GRAM1b G187W)
• Depositing Lab: Yasunori Saheki
• Publication: Koh et al Nat Commun. 2023 Oct 25;14(1):6773. doi: 10.1038/s41467-023-42498-7.

Backbone

• Vector backbone: pLJM1
• Total vector size (bp): 8556
• Modifications to backbone: The PCR product of EGFP-GRAM1b was digested with NheI and MfeI, and then inserted between NheI and EcoRI sites, resulting in the loss of the EcoRI site.
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: NEB® Turbo Competent E. coli can also be used.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: EGFP-tagged GRAM domain of GRAMD1b/Aster-B
• Alt name: EGFP-GRAM1b
• Species: H. sapiens (human)
• Insert Size (bp): 1241
• Mutation: Changed Glycine 187 to Tryptophan
• GenBank ID: NM_020716.4
• Entrez Gene: GRAMD1B (a.k.a. LINC01059)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: EcoRI (destroyed during cloning)
• 5′ sequencing primer: CMV Forward

Resource Information

• Supplemental Documents:
  • D150 pLJM1-EGFP-GRAM-W.gbk

Terms and Licenses

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