pE-SUMO CRYGD
(Plasmid #80755)

• Purpose: Expression of human gamma D-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• Publication: Larry David lab plasmids (unpublished)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: CRYGD
• Species: H. sapiens (human)
• Insert Size (bp): 522
• GenBank ID: HQ258094
• Entrez Gene: CRYGD (a.k.a. CACA, CCA3, CCP, CRYG4, CTRCT4, PCC, cry-g-D)
• 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 HsCD00404130

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 D sequence so it matched the sequence of the mature protein. However, the SUMO tag is resistant to ulp1 SUMO protease cleavage.