pE-SUMO CRYBB3
(Plasmid #80750)

• Purpose: Expression of human beta B3-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• Publication: Larry David lab plasmids (unpublished)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: CRYBB3
• Species: H. sapiens (human)
• Insert Size (bp): 633
• GenBank ID: CR456427
• Entrez Gene: CRYBB3 (a.k.a. CATCN2, CRYB3, CTRCT22)
• 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 plasmid HsCD00620837

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 SUMO tag can be removed using ulp1 SUMO protease to yield the N-terminal Met of mature beta B3-cystallin. Both the SUMO tag and ulp1 SUMO protease can then be removed using an NTA resin. ulp1 SUMO protease can be ordered from LifeSensors or expressed using AddGene plasmid 64697.