pRGEB32-BAR
(Plasmid #126072)

• Purpose: (Empty Backbone) For CRISPR/Cas9 delivery in maize. Expresses Cas9 with rice ubiquitin promoter, BAR with 35S promoter, and sgRNA/PTG with rice snoRNA U3 promoter (pol III). Can produce 1 or more gRNAs.
• Depositing Lab: Charles Hunter
• Publication: Hunter et al Plants (Basel). 2021 Apr 8;10(4). pii: plants10040723. doi: 10.3390/plants10040723.

Backbone

• Vector backbone: pRGEB32
• Backbone manufacturer: Kabin Xie
• Modifications to backbone: Replaced hygromycin selection gene (HPTII) with BAR gene for use in maize.
• Vector type: Plant Expression, CRISPR
• Promoter: 35S for BAR; Rice UBI for Cas9; Rice snoRNA U3 for gRNA/PTG
• Selectable markers: Basta ; BAR

Growth in Bacteria

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

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: M13R(-48)
• 3′ sequencing primer: M13F(-20)

Resource Information

• Supplemental Documents:
  • pRGEB32Bar.gb
• A portion of this plasmid was derived from a plasmid made by: pRGEB32, originally constructed by Kabin Xie from Yinong Yang's lab, was obtained from Addgene (plasmid # 63142).

Terms and Licenses

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

Depositor Comments

This vector has been redesigned for use in Zea mays. Published protocols for using pRGEB32 should be used for this vector.