pUC-ODN-hsRPL3-CterTwStrep-ΔPAM62&56
(Plasmid #208645)

• Purpose: Remove PAM region in HomoArms by introduce SNP rs7495557 and rs760194863, to prevent cleavage after success edit. Digest with nt.BspQI and nb.BsrDI to get ssDNA.
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pUC-ODN
• Modifications to backbone: refer to (Addgene#208642)
• Vector type: CRISPR ; ssDNA donnor preparation for knock-in of RPL3 C-ter tag

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: Twin Strep tag with RPL3 genome HomoArm
• Alt name: Donnor DNA for CRISPR tagging of RPL3
• Species: H. sapiens (human), Synthetic
• Mutation: Two SNP (rs749555789, rs760194863) were introduced to abolish the PAM of sgRNA in (Addgene#208647 and #208979)
• GenBank ID: 5'HomoArm: NC_000022.11:39313122~39312943 3'HomoArm: NC_000022.11:39312939~39312773
• Entrez Gene: RPL3 (a.k.a. ASC-1, L3, TARBP-B, uL3)
• Tag / Fusion Protein:
  • Twin Strep

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: M13-F
• 3′ sequencing primer: M13-R

Resource Information

• Supplemental Documents:
  • pUC-ODN-hsRPL3-CterTwStrep-ΔPAM62&56.gb

Terms and Licenses

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