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Purpose(Empty Backbone) modified pENTR vector for N-terminal 3x HA tag fusion with your gene of interest
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 17456 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepENTR1A
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Backbone manufacturerInvitrogen
- Backbone size (bp) 2427
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Modifications to backboneA short cloning site region and stop codon replaced ccdB gene in pENTR1A. Inserted 3x HA tag
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Vector typeMammalian Expression, Bacterial Expression, Yeast Expression, Insect Expression ; modified Gateway Entry vector
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameNone
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Tag
/ Fusion Protein
- 3xHA (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') (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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 3x HA tagged Entry vector. 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: EU334817
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pE2n was a gift from Giovanna Benvenuto (Addgene plasmid # 17456 ; http://n2t.net/addgene:17456 ; RRID:Addgene_17456) -
For your References section:
A modified Gateway cloning strategy for overexpressing tagged proteins in plants. Dubin MJ, Bowler C, Benvenuto G. Plant Methods. 2008 Jan 22. 4(1):3. 10.1186/1746-4811-4-3 PubMed 18211686