pSLIK_Hyg_mTurquoise2_Atg4BC74A
(Plasmid #161733)

• Purpose: Doxycycline-inducible expression of Atg4B(C74A) fused with mTurquoise2 to inhibit autophagy
• Depositing Lab: Alec Kimmelman
• Publication: Yamamoto et al Nature. 2020 May;581(7806):100-105. doi: 10.1038/s41586-020-2229-5. Epub 2020 Apr 22.

Backbone

• Vector backbone: pSLIK-hygro
• Backbone manufacturer: Addgene #25737
• Backbone size w/o insert (bp): 12262
• Total vector size (bp): 14694
• Vector type: Mammalian Expression, Lentiviral ; destinatioin vector for gateway cloning
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: After plasmid purification, please make sure your plasmid contains 7 repeats of TetO sequence as this region can occasionally be lost by recombination.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Atg4B(C74A) (Atg4b Mouse)
• Alt name: Atg4B
• Species: M. musculus (mouse)
• Mutation: 220-222 TGC (Cys) is changed to GCA (Ala)
• GenBank ID: NM_174874
• Entrez Gene: Atg4b (a.k.a. 2510009N07Rik, Apg4b, Atg4bl, Autl1, MmAPG4B)
• Tag / Fusion Protein:
  • mTurquoise2 (N terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: attB1
• 3′ sequencing primer: attB2

Resource Information

• Supplemental Documents:
  • Annotated Depositor's Sequence (.dna)
  • Annotated Depositor's Sequence (.gb)

Terms and Licenses

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