pcDNA3 c-SRC (250-536) K298M HA
(Plasmid #42210)

• Depositing Lab: Robert Lefkowitz
• Publication: Miller et al J Biol Chem. 2000 Apr 14;275(15):11312-9.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5446
• Total vector size (bp): 7000
• 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: c-SRC (250-536) HA
• Alt name: SRC
• Alt name: sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog
• Alt name: p60-Src
• Species: H. sapiens (human)
• Mutation: contains amino acids 250-536; K298M
• GenBank ID: NM_005417.3 AAA60584.1
• Entrez Gene: SRC (a.k.a. ASV, SRC1, THC6, c-SRC, p60-Src)
• Promoter: CMV
• Tag / Fusion Protein:
  • HA (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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

Depositor Comments

All c-SRC mutants were generated by polymerase chain reaction using c-SRC (WT) (Addgene Plasmid #42202) as a template, and polymerase chain reaction products containing a minimal Kozak sequence were cloned into the HindIII and XhoI sites of pcDNA3. This construct contains amino acids 250-536 of human c-SRC and the kinase inactivating Lys to Met mutation at amino acid 298 followed by a carboxyl-terminal HA tag. All sequences were confirmed using an automated ABI DNA sequencer (Howard Hughes Nucleic Acid Facility, Duke University).

Alternate plasmid name: pcDNA3 c-SRC-SH1(KD)