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PurposeExpress eGFP reporter for HDR-induced knock-in
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Depositing Lab
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Sequence Information
Ordering
This material is available to academics and nonprofits only.
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 87865 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonepsuper puro
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Backbone manufacturer4353
- Total vector size (bp) 6478
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Modifications to backboneDNA fragment carrying ires-eGFP was amplified from plasmid pLenti6.3-MCS-IRES2-EGFP (Life Technologies). The fragment obtained was inserted into BamHI and XhoI sites in pSuper-puro vector. Two homology arms were amplified from GAPDH genomic locus, and inserted into BamHI and MfeI sites at 5’ and Sal1 and HpaI sites at 3’ of the IRES-eGFP fragment.
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Vector typeHomologous recombination for GAPDH loci
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 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 nameeGFP
Resource Information
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Article Citing this Plasmid
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.
How to cite this plasmid
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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:
ires-eGFP(+HAs) donor-1 was a gift from Bo Feng (Addgene plasmid # 87865 ; http://n2t.net/addgene:87865 ; RRID:Addgene_87865) -
For your References section:
Knock-in of large reporter genes in human cells via CRISPR/Cas9-induced homology-dependent and independent DNA repair. He X, Tan C, Wang F, Wang Y, Zhou R, Cui D, You W, Zhao H, Ren J, Feng B. Nucleic Acids Res. 2016 May 19;44(9):e85. doi: 10.1093/nar/gkw064. Epub 2016 Feb 4. 10.1093/nar/gkw064 PubMed 26850641
