pET28a-HIS-TagGFP2-TEV-GGG-ELP(40nm)-Cys
(Plasmid #90474)

• Purpose: Elastin Like Polypeptide: [(VPGEG)-(VPGVG)4-(VPGAG)2-(VPGGG)2-(VPGEG)]2, with N-Terminal TagGFP2 and TEV-cleavage site, leaving an N-terminal Sortase Recognition Sequence GGG and a C-terminal Cysteine
• Depositing Lab: Hermann Gaub
• Publication: Gaub Lab ELP Plasmids (unpublished)

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5216
• Total vector size (bp): 6293
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For protein production transformed NiCo21(DE3) strains in auto induction media (supplemented with 5 mg/ml proline, valine and 10 mg/ml glycin) were grown for 24 hrs (6 hrs 37°C, 18 hrs 25°C).
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: TagGFP2-TEV-GGG-ELP(40nm)-Cys
• Alt name: Elastin like polypeptide
• Species: Synthetic
• Insert Size (bp): 1077
• Promoter: T7
• Tags / Fusion Proteins:
  • Sortase-Tag (N terminal on insert)
  • Cysteine (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CCGATCTTCCCCATCGGT
• 3′ sequencing primer: CTAGTTATTGCTCAGCGGT

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The underlying cloning procedure of the employed ELP genes is described in this article:
Ott, W.; Nicolaus, T.; Gaub, H. E.; Nash, M. A. Sequence-Independent Cloning and Post-Translational Modification of Repetitive Protein Polymers through Sortase and Sfp-Mediated Enzymatic Ligation. Biomacromolecules 2016, 17, 1330−1338.