pPICZA_CeTRIC-B1_Del
(Plasmid #87344)

• Purpose: Inducible expression of CeTRIC-B1 in P.Pichia strains
• Depositing Lab: Zhenfeng Liu
• Publication: Yang et al Nature. 2016 Oct 27;538(7626):537-541. doi: 10.1038/nature19767. Epub 2016 Oct 3.

Backbone

• Vector backbone: pPICZ A
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 3221
• Total vector size (bp): 3977
• Vector type: Yeast Expression
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Bleocin (Zeocin), 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TRIC-B1
• Alt name: TMEM38B
• Species: C. elegans (nematode)
• Insert Size (bp): 756
• Mutation: Δ248-297
• GenBank ID: 290457497
• Promoter: AOX1
• Tag / Fusion Protein:
  • polyhistidine (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XhoI (destroyed during cloning)
• 5′ sequencing primer: 5' AOX1
• 3′ sequencing primer: 3' AOX1

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pPICZA_TRIC-B1 was synthesized by Genscript.

Terms and Licenses

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

Depositor Comments

Vector pPICZ A and insert sequence were modified as follows:
1. Restriction sites including Pml I; Sfi I; BsmB I; Asp718 I; Kpn I were destroyed during cloning full-length CeTRIC-B1 to the vector.
2. In the C-terminal truncation, the sequence between XhoI and polyhistidine had been deleted. That is to say, restriction sites including Xho I, Sac II, Not I, Apa I, and myc-epitope, were destroyed during truncation.
3. A kozak suqence "ACC" was inserted between the restriction site "EcoR I" and initial codon, we added these 3 bp into insert sequence size.

In summary, the size of pPICZ A backbone is changed from 3329 bp to be 3221 bp.