PB-PE SpG
(Plasmid #179081)

• Purpose: NOTE: An improved version of this plasmid is now available. See Addgene plasmid #242072. All-in-one prime editor piggyBac transposon, SpG variant
• Depositing Lab: Tobias Cantz
• Depositing Organization: Medizinische Hochschule Hannover
  Located in Germany
• Publication: Eggenschwiler et al Sci Rep. 2021 Nov 12;11(1):22154. doi: 10.1038/s41598-021-01689-2.

Backbone

• Vector backbone: PB-PE
• Backbone manufacturer: selfmade
• Backbone size w/o insert (bp): 11803
• Total vector size (bp): 13136
• Modifications to backbone: addition of 3' and 5' piggyBac repeats, addition of pU6-pegRNA(B12B) sequence, addition of CAG promoter, addition of bGH (pA)
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology ; piggyBac transposon
• Selectable markers: Puromycin ; thymidine kinase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: It is highly recommended to use NEB Stable at 30°C. Cloning efficiency and stability of plasmid can be compromised when using other strains / higher temperatures!
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: SpCas9 SpG_H840A-PE2-MMLV-RT(dBB)-P2A-PAC_dTK(dBB)
• Species: Synthetic
• Insert Size (bp): 8016
• Mutation: D1135L, S1136W, G1218K, E1219Q, R1335Q, T1337R
• Promoter: CAG
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: GCTGTCTCATCATTTTGGC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: 1378 bp of Cas9 SpG was PCR amplified from pCMV-PE2-SpG (Addgene Plasmid #159978)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

NOTE: An improved version of this plasmid is now available: https://www.addgene.org/242072/.

DNA (Catalog # 179081-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 179081-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.