pEF6-Lck-TurboID
(Plasmid #159433)

• Purpose: expresses Lck-TurboID in mammalian cells
• Depositing Lab: Arthur Salomon
• Publication: Chua et al J Proteome Res. 2020 Nov 13. doi: 10.1021/acs.jproteome.0c00616.

Backbone

• Vector backbone: pEF6
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5818
• Total vector size (bp): 8249
• Vector type: Mammalian Expression
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Lck
• Alt name: Lck tyrosine kinase
• Species: H. sapiens (human)
• Insert Size (bp): 1527
• GenBank ID: NM_001042771.2
• Entrez Gene: LCK (a.k.a. IMD22, LSK, YT16, p56lck, pp58lck)
• Promoter: EF-1a

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (unknown if destroyed)
• 3′ cloning site: BbvCI (unknown if destroyed)
• 5′ sequencing primer: TATATGAATTCGCCATGGGCTGTGGCTGC
• 3′ sequencing primer: ATATGTTTAAACTCAAGCGTAATCTGGAACATCGTATGGGTAAGGCTGAGGCTGGTACTG

Gene/Insert 2

• Gene/Insert name: TurboID (BirA mutant)
• Species: E. coli
• Insert Size (bp): 969
• Mutation: Q65P, I87V, R118S, E140K, Q141R, S150G, L151P, V160A, T192A, K194I, M209V, M241T, S263P, I305V
• Tag / Fusion Protein:
  • HA (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbvCI (unknown if destroyed)
• 3′ cloning site: PmeI (unknown if destroyed)
• 5′ sequencing primer: ATATGTTTAAACTCAAGCGTAATCTGGAACATCGTATGGGTAAGGCTGAGGCTGGTACTG
• 3′ sequencing primer: TATATGTTTAAACTTAAGCGTAATCTGGAACATCGTATGGGTATCCGTCCAGGGTCAGGCGCTCC

Resource Information

• Supplemental Documents:
  • pEF6 Lck TurboID HA.gbk
• A portion of this plasmid was derived from a plasmid made by: TurboID obtained from Addgene #107169 (Alice Ting Lab)

Terms and Licenses

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