pGKTM2SP00414
(Plasmid #258605)

• Purpose: Level 2 all-in-one R2 editor: R2Tg (T. guttata R2 retroelement, 2 introns); intron-containing mCherry retrotransposition reporter; mTagBFP2 marker; pCsB spectinomycin-resistant plant binary vector.
• Depositing Lab: Gözde Demirer
• Publication: Muchenje et al Nat Biotech. 2026 Jun 19.

Backbone

• Vector backbone: pCsB binary vector
• Backbone manufacturer: Dr. Nicola J. Patron
• Backbone size w/o insert (bp): 6667
• Total vector size (bp): 16718
• Vector type: Plant Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: pVmv-R33-wt R2Tg 5′ UTR-t35s-mCherry (A. thaliana intron)-TMV 5′ UTR-p35s-wt R2Tg 3′ UTR-R4-HDV-tHSP
• Species: Synthetic
• Insert Size (bp): 3108
• Promoter: pVmv
• Tag / Fusion Protein:
  • Bipartite nuclear localization sequence (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Golden Gate
• 5′ sequencing primer: Whole plasmid sequencing

Gene/Insert 2

• Gene/Insert name: pCmY-R2Tg[+2 A. thaliana introns]-tHSP
• Species: Taeniopygia guttata
• Insert Size (bp): 4529
• Promoter: pCmY
• Tag / Fusion Protein:
  • bipartite nuclear localization sequence (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Golden Gate
• 5′ sequencing primer: Whole plasmid sequencing

Gene/Insert 3

• Gene/Insert name: pCmY-mTagBFP2-tHSP
• Species: Synthetic
• Insert Size (bp): 2130
• Promoter: pCmY
• Tag / Fusion Protein:
  • mTagBFP2 (C terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Golden Gate
• 5′ sequencing primer: Whole plasmid sequencing

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please visit https://doi.org/10.1101/2025.08.22.671877 for bioRxiv preprint.