pET28a(+)v2_TwinStrep-SUMO-φC31(PhiC31)-8xHis
(Plasmid #246982)

• Purpose: Expresses E. coli codon-optimized TwinStrep-SUMO-tagged φC31(PhiC31) integrase with N- and C-terminal NLS, using an improved pET design from Shilling et al. 2020.
• Depositing Lab: Amro Hamdoun
• Publication: Sea Urchin Transgenesis Reagent Constructs (unpublished)

Backbone

• Vector backbone: pET28a(+)v2
• Backbone manufacturer: NA
• Backbone size w/o insert (bp): 5218
• Total vector size (bp): 7627
• Vector type: Bacterial Expression, Synthetic Biology

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: φC31(PhiC31)
• Alt name: phage phi-C31 integrase
• Insert Size (bp): 2403
• GenBank ID: NC_001978
• Promoter: T7
• Tags / Fusion Proteins:
  • TwinStrep (N terminal on insert)
  • SUMO (N terminal on insert)
  • SV40 (N terminal on insert)
  • Nucleoplasmin (C terminal on insert)
  • 8xHis (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: NA

Resource Information

• Supplemental Documents:
  • pET28a(_)v2_TwinStrep-SUMO-φC31(PhiC31)-8xHis.gb
• A portion of this plasmid was derived from a plasmid made by: The original gene sequence for φC31(PhiC31) integrase was obtained from Addgene Plasmid #68310. The sequence and all tags were codon optimized for E. coli expression and synthesized by Twist Bioscience. The original backbone sequence for this modified pET28a(+) was obtained from Addgene Plasmid #154464.

Terms and Licenses

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

Depositor Comments

We use a tag-free SUMO protease (ulp1) for cleaving the TwinStrep tag from the final product. Tag-free ulp1 can be synthesized using Addgene Plasmid #190063 and a His-tagged TEV protease or HRV-3C protease (PreScission protease).

The total insert size displayed on this page (2403bp) includes all tags and linkers.