PE-SUMO CRYBA4
(Plasmid #217671)

• Purpose: For expression of SUMO tagged human beta A4 crystallin
• Depositing Lab: Larry David
• Publication: Completion of previous SUMO tagged human crystallin submission plus plasmids for native chemical ligation to produced gamma S crystallin (unpublished)

Backbone

• Vector backbone: PE-SUMO
• Backbone manufacturer: LifeSensors
• Backbone size w/o insert (bp): 5761
• Total vector size (bp): 6323
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: CRYBA4
• Species: H. sapiens (human)
• Insert Size (bp): 585
• GenBank ID: CRYBA4
• Entrez Gene: CRYBA4 (a.k.a. CTRCT23, CYRBA4, MCOPCT4)
• Promoter: T7
• Tags / Fusion Proteins:
  • 6XHis (N terminal on backbone)
  • Smt3 (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Bsal (unknown if destroyed)
• 3′ cloning site: Xbal (unknown if destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 term

Resource Information

• Supplemental Documents:
  • pE-SUMO CRYBA4.gbk
• A portion of this plasmid was derived from a plasmid made by: DNASU HsCD00620816

Terms and Licenses

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

Depositor Comments

Expressed in an insoluble state and must recovered from the insoluble pellet of lysed bacteria using 8M urea. N-terminal tag can still be removed using ULP-1 after diluting urea. The N-terminal methionine has been removed so that the protein mirrors the state found in the lens that is missing its N-terminal methionine.