pE4n
(Plasmid #17458)

• Purpose: (Empty Backbone) modified pENTR vector for N-terminal GST tag fusion with your gene of interest
• Depositing Lab: Giovanna Benvenuto
• Publication: Dubin et al Plant Methods. 2008 Jan 22. 4(1):3.

Backbone

• Vector backbone: pENTR1A
• Backbone manufacturer: Invitrogen
• Backbone size (bp): 2982
• Modifications to backbone: A short cloning site region and stop codon replaced ccdB gene in pENTR1A. Inserted GST tag and thrombin cleavage site
• Vector type: Mammalian Expression, Bacterial Expression, Worm Expression ; modified Gateway Entry vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • Thrombin site (N terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: pENTR-F (5'-CTACAAACTCTTCCTGTTAGTTAG-3')
• 3′ sequencing primer: pENTR-R (5'-ATGGCTCATAACACCCCTTG-3')

Terms and Licenses

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

Depositor Comments

Part of a modified Gateway Entry vector system for generating epitope tagged constructs. Clone your gene of interest into this vector using the BamHI and NotI cloning sites to obtain a N-terminal GST tagged Entry vector with a thrombin cleavage site for protein purification purposes. The resulting Entry vector can be recombined into Gateway Destination vectors for bacteria, insect mammalian, plant or yeast expression via the Gateway LR reaction.

The tag is present in the Entry cassette to avoid the presence of att-derived protein linker sequences between the tag and gene or interest corresponding to the attB recombination site. A stop codon is already present within the vector (see plasmid map).

GenBank accession number: EU334819