pKb-TnpB2
(Plasmid #215334)

• Purpose: ISDra2 expression driven by PolII promoter ; 20 bp guide RNA can be cloned in BsaI site
• Depositing Lab: Kutubuddin Molla
• Publication: Karmakar et al Plant Biotechnol J. 2024 Jun 28. doi: 10.1111/pbi.14416.

Backbone

• Vector backbone: pRGEB32
• Backbone manufacturer: Yinong Yang
• Backbone size w/o insert (bp): 9823
• Total vector size (bp): 14975
• Modifications to backbone: 1. Replaced gRNA scaffold with HH ribozyme-reRNA-gRNA cloning site-HDV ribozyme. 2. Replaced OsU3 promoter with ZmUbi promoter. 3. Replaced Cas9 with rice codon optimized TnpB nuclease.
• Vector type: Plant Expression
• Selectable markers: Hygromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Rice codon optimized ISDra2 TnpB
• Species: Deinococcus radiodurans
• Insert Size (bp): 1224
• Promoter: Rice ubiquitin promoter for TnpB and maize ubiquitin promoter for HH ribozyme-reRNA-guide RNA-HDV ribozyme expression
• Tag / Fusion Protein:
  • Not applicable

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (unknown if destroyed)
• 3′ cloning site: SacI (unknown if destroyed)
• 5′ sequencing primer: M13R
• 3′ sequencing primer: M13F

Terms and Licenses

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