pLX304-BCL-XL-499/500R
(Plasmid #203575)

• Purpose: Expresses V5-tagged BCL-XL with resistance to shRNA (TRCN0000033499, TRCN0000033500) in mammalian cells.
• Depositing Lab: Abhishek Chakraborty
• Publication: Grubb et al Clin Cancer Res. 2022 Nov 1;28(21):4689-4701. doi: 10.1158/1078-0432.CCR-22-0669.

Backbone

• Vector backbone: pLX304
• Backbone size w/o insert (bp): 9377
• Total vector size (bp): 8428
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Blasticidin

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: BCL2L1
• Alt name: BCL2 like 1
• Species: H. sapiens (human)
• Insert Size (bp): 774
• Mutation: silent point mutations to make resistant shRNA 499/500R (TRCN0000033499, TRCN0000033500)
• GenBank ID: 598
• Entrez Gene: BCL2L1 (a.k.a. BCL-XL/S, BCL2L, BCLX, Bcl-X, PPP1R52)
• Promoter: CMV
• Tag / Fusion Protein:
  • V5-tagged (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: WPRE-R

Terms and Licenses

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

Depositor Comments

BCL-XL was sub-cloned, using gateway cloning, from a BCL-XL-GFP fusion construct obtained from Dr. Ruth Keri's laboratory (CCF). Silent mutations were introduced in the shRNA-recognition sequences for both sh499 and sh500 using Site-directed mutagenesis.