PB-PE SpRY
(Plasmid #179082)

• Purpose: All-in-one prime editor piggyBac transposon, SpRY variant
• Depositing Lab: Tobias Cantz
• 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): 8921
• 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 SpRY_H480A-PE2-MMLV-RT(dBB)-P2A-PAC_dTK(dBB)
• Species: Synthetic
• Insert Size (bp): 8016
• Mutation: A61R, L1111R, D1135L, S1136W, G1218K, E1219Q, N1317R, A1322R, R1333P, 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

• Supplemental Documents:
  • PB-PE SpRY.gbk
• A portion of this plasmid was derived from a plasmid made by: 4260 bp of Cas9 SpRY was PCR amplified from pCMV-PE2-SpRY (Addgene Plasmid #159979)

Terms and Licenses

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