pLJM1-SLC35B2-FLAG
(Plasmid #154863)

• Purpose: Transient or Lentiviral expression of sgRNA (Addgene#154860) resistant SLC35B2
• Depositing Lab: Yihong Ye
• Publication: Zhang et al Proc Natl Acad Sci U S A. 2020 May 19;117(20):10865-10875. doi: 10.1073/pnas.1918617117. Epub 2020 May 4.

Backbone

• Vector backbone: pLJM1-EGFP (Addgene# 19319)
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SLC35B2
• Alt name: PAPST1
• Species: H. sapiens (human)
• Mutation: ctcctggtgcagtacttc => ttattagtccaatatttt
• GenBank ID: NM_178148
• Entrez Gene: SLC35B2 (a.k.a. HLD26, PAPST1, SLL, UGTrel4)
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG (C terminal on insert)

Cloning Information

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

Terms and Licenses

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

Depositor Comments

Please note: Plasmid contains an A291V mutation in SLC35B2 compared to the NCBI reference sequence NM_178148. This mutation is not expected to affect plasmid function as the plasmid was still able to rescue the heparan synthesis and endocytosis defects of SLC35B2 knockout cells.