pCMV-PE5max-SF3B1-K700E
(Plasmid #200317)

• Purpose: Expresses the PE5max system to engineer the SF3B1 K700E mutation. Co-expresses both a puromycin resistance gene and tagRFP for selection.
• Depositing Lab: Lili Wang
• Publication: Fernandez et al Cancer Res Commun. 2024 Aug 28. doi: 10.1158/2767-9764.CRC-24-0145.

Backbone

• Vector backbone: pCMV
• Total vector size (bp): 15245
• Vector type: Mammalian Expression
• Selectable markers: Puromycin ; RFP

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: K700E-ng1+pg2
• Alt name: K700E-ng1+pg2-Puro-2A-RFP
• Species: H. sapiens (human); Will only work on human SF3B1
• Insert Size (bp): 3120

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SgrDI (not destroyed)
• 3′ cloning site: SgrDI (not destroyed)
• 5′ sequencing primer: Whole plasmid sequencing with Primordium.
• 3′ sequencing primer: Whole plasmid sequencing with Primordium.

Gene/Insert 2

• Gene/Insert name: PEmax-2A-MLH1dn
• Species: H. sapiens (human)
• Insert Size (bp): 8715
• Promoter: CMV

Cloning Information for Gene/Insert 2

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7-forward
• 3′ sequencing primer: BGH-reverse

Resource Information

• Supplemental Documents:
  • pCMV-PE5Max-SF3B1-K700E.gb

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.1101/2024.02.23.581842 for bioRxiv preprint.