pSC-PreE2
(Plasmid #52264)

• Purpose: Bacterial expression of GST-tagged TARGET proteins fused to SUMO-E2 with PreScission protease cleavage site linker
• Depositing Lab: Primo Schaer
• Publication: Weber et al PLoS One. 2014 Jul 9;9(7):e102157. doi: 10.1371/journal.pone.0102157. eCollection 2014.

Backbone

• Vector backbone: pTXB3
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 6707
• Total vector size (bp): 7083
• Modifications to backbone: GyrA-intein and CBD domain replaced by expression unit with the PreScission cleavage site fused to Ubc9 and GST
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: In combination with a second vector for protein expression in BL21(DE3) cells, the selection pressure was reduced by half to 50 mg/L Ampicillin
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ubc9 GST fusion
• Alt name: Ube2l
• Alt name: Ubc9
• Species: H. sapiens (human)
• Insert Size (bp): 1773
• GenBank ID: NM_003336.1
• Entrez Gene: UBE2I (a.k.a. C358B7.1, P18, UBC9)
• Tag / Fusion Protein:
  • GST (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: BglII (destroyed during cloning)
• 5′ sequencing primer: TAATACGACTCACTATAGGG
• 3′ sequencing primer: TGCTAGTTATTGCTCAGCGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Ubc9 cDNAs was amplified from pGEX-based expression vector kindly gifted by Ronald Hay

Terms and Licenses

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