pJK_proK1_eGFP
(Plasmid #59853)

• Purpose: Constitutively expresses eGFP with proK1 promoter
• Depositing Lab: Timothy Whitehead
• Publication: Bienick et al PLoS One. 2014 Oct 6;9(10):e109105. doi: 10.1371/journal.pone.0109105. eCollection 2014.

Backbone

• Vector backbone: pET-29b(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 3582
• Total vector size (bp): 4296
• Modifications to backbone: Deletion of lacI, lacO, and T7 promoter by cloning proB in-between BglI and XbaI. Removal of the kanamycin resistance gene and insertion of the beta-lactamase (AmpR) gene from pET-22b.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Enhanced Green Fluorescent Protein
• Alt name: eGFP
• Alt name: BBa_E0040
• Alt name: GFPmut3
• Species: Aequorea victoria
• Insert Size (bp): 714
• Mutation: S2R,S65G,S72A
• Promoter: proK1
• Tag / Fusion Protein:
  • 6x-His (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: GGTGATTCATTCTGCTAA
• 3′ sequencing primer: T7 Term

Terms and Licenses

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