Bcr/Abl P190-pLEF
(Plasmid #38159)

• Depositing Lab: Nora Heisterkamp
• Publication: Kweon et al J Biol Chem. 2008 Feb 8;283(6):3023-30. Epub 2007 Dec 10.

Backbone

• Vector backbone: pLEF
• Backbone size w/o insert (bp): 7400
• Total vector size (bp): 14000
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: BCR/ABL P190
• Species: H. sapiens (human)
• Insert Size (bp): 7000
• Mutation: T117M in c-ABL1
• Entrez Gene: ABL1 (a.k.a. ABL, BCR-ABL, CHDSKM, JTK7, bcr/abl, c-ABL, c-ABL1, p150, v-abl)
• Entrez Gene: BCR (a.k.a. ALL, BCR1, CML, D22S11, D22S662, PHL)
• Promoter: SV40
• Tag / Fusion Protein:
  • GST (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: GST or T7
• 3′ sequencing primer: SP6

Terms and Licenses

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

Depositor Comments

Constructed by Suparna Mishra. Generated in a 3-way ligation consisting of 5' 0.75 kb BamHI/Sal I + 3' 6.3 kb Sal I/EcoRI BCR/ABL fragments into pLEF digested with BamHIxEcoRI. The ABL part of the insert contains the natural TAG and 3' UT sequences.

Addgene NGS identified a sequence discrepancy at nucleotide 3168 resulting in a T117M mutation in the ABL-1 sequence, relative to Genbank ID NP_005148.2 and Addgene Plasmid #38158. This is not a known polymorphism and the effect of this, if any, on the protein is not known.