pKLV2-U6gRNA5(hPLK1)-PGKpuro2ABFP-W
(Plasmid #163138)

• Purpose: Lentiviral gRNA plasmid targeting human PLK1 gene, co-expression of BFP tag
• Depositing Lab: Sok Ching Cheong
• Publication: Chai et al Elife. 2020 Sep 29;9. pii: 57761. doi: 10.7554/eLife.57761.

Backbone

• Vector backbone: pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W
• Total vector size (bp): 8650
• Modifications to backbone: Used BbsI enzyme to clone in gRNA targeting human PLK1 gene
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: PLK1
• gRNA/shRNA sequence: GTCGGACGCGGACACCAAGG
• Species: H. sapiens (human)
• Entrez Gene: PLK1 (a.k.a. PLK, STPK13)
• Promoter: U6
• Tag / Fusion Protein:
  • BFP

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbsI (destroyed during cloning)
• 3′ cloning site: BbsI (destroyed during cloning)
• 5′ sequencing primer: AGATAATTAGAATTAATTTGACTG

Resource Information

• Supplemental Documents:
  • Kosuke Yusa - cloning protocol used to generate the plasmid from pKLV2-U6gRNA5-PGKpuro2ABFP-W

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry