pET29b(+)_NdeI-NcoI_SUMO(21-98)_CV
(Plasmid #254142)

• Purpose: Vector for cloning in inducible amphipathic helix of choice to the N-terminal of the AH-SUMO platform. Clone using HiFi assembly between NdeI-NcoI and add "GSGSMSDSEVNQEAKPEVKPEVKP" to end of peptide.
• Depositing Lab: Peter Chung
• Publication: Margaritakis et al Commun Chem. 2026 Apr 2;9(1):165. doi: 10.1038/s42004-026-01994-9.

Backbone

• Vector backbone: pET29b(+)
• Backbone size w/o insert (bp): 5237
• Total vector size (bp): 5537
• Vector type: Bacterial Expression

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: NcoI_SMT3-SUMO(21-98)_C_sTEV_His6
• Alt name: SMT3
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 300
• Mutation: Contains encoding region for enzyme site NdeI, then followed by the 21-98 AA of SMT3_yeast, followed by encoding region for a Cystein, a TEV site and a hexa-histidine encoding region.
• GenBank ID: NP_010798.1
• Entrez Gene: SMT3 (a.k.a. YDR510W)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Twistbioscience

Terms and Licenses

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

Depositor Comments

This vector is a modified pET29b(+) vector that includes one added cloning site (NcoI) followed by the encoding region for a fragment of the SMT3_yeast gene (21-98) followed by a TEV site (ENLYFQS) and a hexa-histidine site (HHHHHH*). Any peptide of interest can be cloned in the vector using the two cloning sites (NdeI and NcoI) via HiFi assembly. When adding a peptide of interest this way, the remaining N-terminal portion of the SMT3 gene should be added as well, separated by the peptide of interest with a GSGS encoding region (GSGSMSDSEVNQEAKPEVKPEVKP). This can result in a vector which encodes for the expression of a recombinant gene of the following type : PEPTIDE-GSGS-SUMO(1-98)-C-sTEV_His6*.

Please visit https://doi.org/10.1101/2025.05.12.653516 for the bioRxiv preprint.