pSBbi_RP_3XNLS-MicroID2-FLAG
(Plasmid #188523)

• Purpose: Encodes the biotin proximity labeling enzyme MicroID2 with a 3X nuclear localization sequence (NLS) for expression and stable integration into mammalian cells
• Depositing Lab: James Londino
• Publication: Johnson et al Mol Cell Proteomics. 2022 Jun 7:100256. doi: 10.1016/j.mcpro.2022.100256.

Backbone

• Vector backbone: pSBbi-RP
• Backbone manufacturer: Eric Kowarz Lab
• Backbone size w/o insert (bp): 6350
• Total vector size (bp): 7056
• Vector type: Mammalian Expression ; Transposon mediated gene integration
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: 3XNLS-MicroID2
• Species: Synthetic; Aquifex aeolicus
• Insert Size (bp): 652
• Promoter: EF1a/RPBSA
• Tag / Fusion Protein:
  • FLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SfiI/NcoI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: EF1a-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Supplemental Documents:
  • pSBbi_RP_3XNLS-microID2-FLAG.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry