pcDNA3.1-CRISPR-Puro-sgRPL3-62
(Plasmid #208647)

• Purpose: pcDNA3.1 based plasmid for transient transfection of sgRNA-cas9. Containing sgRNA targeting human RPL3 (GRCh38.p14_Chr22:39312968-39312987), 62 is a number given by www.e-crisp.org.
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pcDNA3.1-CRISPR-Puro
• Modifications to backbone: refer to (Addgene#208646)
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Neomycin (select with G418), Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: We use stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: sgRNA targeting human RPL3 (ENST00000216146) C-terminal, No.62
• Alt name: RPL3-sgR62
• gRNA/shRNA sequence: TCTTTCTTCAGTGGTCCCTG--/PAM/tGG/
• Species: H. sapiens (human), Synthetic
• GenBank ID: NC_000022.11:39312968-39312987
• Entrez Gene: RPL3 (a.k.a. ASC-1, L3, TARBP-B, uL3)
• Promoter: CMV and U6 in tandem

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBI-v2 (destroyed during cloning)
• 3′ cloning site: BsmBI-v2 (destroyed during cloning)
• 5′ sequencing primer: CMV-F, Human-U6-promoter-F

Resource Information

• Supplemental Documents:
  • pcDNA3.1-CRISPR-Puro-sgRPL3-62.gb

Terms and Licenses

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

Depositor Comments

co-transfect with pcDNA3.1-CRISPR-Puro-sgRPL3-56 (Addgene#208979) for better knock-in, single cut not perfermed good for knock-in.