pPPEDs
(Plasmid #162469)

• Purpose: protoplast test vector of pPPED
• Depositing Lab: Adam Bogdanove
• Publication: Wang et al Front Genome Ed. 2021 Mar 4;3:617553. doi: 10.3389/fgeed.2021.617553. eCollection 2021.

Backbone

• Vector backbone: pBS(-)
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 2958
• Total vector size (bp): 13521
• Modifications to backbone: Add 2x35S promoter, Arabidopsis codon optimized Cas9H840A, MLV reverse transcriptase, NOS terminator and gateway cloning attR site.
• Vector type: Plant Expression ; Prime editing destination vector for transient expression in dicot plant protoplast

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Cas9H840A
• Species: Synthetic
• Insert Size (bp): 10576
• Mutation: Change histidine 840 to Alanine
• Tag / Fusion Protein:
  • MLV reverse transcriptase (C terminal on insert)

Gene/Insert 2

• Gene/Insert name: attR gateway destination cassette
• Species: Synthetic

Resource Information

• Supplemental Documents:
  • pPPEDs.gb
• A portion of this plasmid was derived from a plasmid made by: Dr. Ning Zhang from Dr. Greg Martin's lab.

Terms and Licenses

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

Depositor Comments

This vector was modified from pPPED, which is made by Dr. Ning Zhang from Dr. Greg Martin lab.
reference:
1. Targeted genome modifications in soybean with CRISPR/Cas9.
Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA.
BMC Biotechnology. 2015;15:16.
Please visit https://www.biorxiv.org/content/10.1101/2020.07.16.206276v1 for bioRxiv preprint.