pE-SUMO CRYGS
(Plasmid #80753)

• Purpose: Expression of human gamma S-crystallin in E. coli
• Depositing Lab: Larry David
• Publication: Larry David lab plasmids (unpublished)

Backbone

• Vector backbone: pE-SUMO
• Backbone manufacturer: LifeSensors
• Backbone size w/o insert (bp): 5761
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: CRYGS
• Species: H. sapiens (human)
• Insert Size (bp): 531
• GenBank ID: BC069478
• Entrez Gene: CRYGS (a.k.a. CRYG8, CTRCT20)
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHIS (N terminal on insert)
  • Smt3 SUMO (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: XbaI (destroyed during cloning)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • annotated sequence
• A portion of this plasmid was derived from a plasmid made by: DNASU HsCD00620006

Terms and Licenses

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

Depositor Comments

Expresses well as a N-terminal SUMO tagged protein with a 6XHis tag on the N-terminus for purification. The N-terminal Met was removed from the gamma S sequence so it matched the sequence of the mature protein. Unlike gamma A, B, C, ad D, the SUMO tag is easily removed by ulp1 SUMO protease cleavage. ulp1 SUMO protease can be ordered from LifeSensors or expressed using Addgene plasmid 64697.