|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||86271||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameHMG20A homology arms
SpeciesH. sapiens (human)
Entrez GeneHMG20A (a.k.a. HMGX1, HMGXB1)
/ Fusion Protein
- 3XFLAG-P2A-NeoR (C terminal on insert)
- Cloning method Gibson Cloning
- 5′ sequencing primer acgcctgtgaaaccgtacta
- 3′ sequencing primer aactgttgggaagggcgatc (Common Sequencing Primers)
Terms and Licenses
This plasmid was co-transfected along with PX458_HMG20A_1 and PX458_HMG20A_2 into HepG2 cells, and genomic editing was verified by PCR. The donor nucleotide sequence in the final codon of the HMG20A gene is modified to discourage sgRNA recognition, while retaining the native amino acid sequence. We performed ChIP-seq on this cell line using anti-FLAG Sigma F1804 and obtained data that passed our quality tests. Immunoprecipitation with anti-FLAG Sigma F1804 followed by mass spectrometry identified HMG20A. All released data, including ChIP-seq files and validation data, are available at the ENCODE web portal: https://www.encodeproject.org/search/?searchTerm=flag&type=Experiment&lab.title=Richard+Myers%2C+HAIB . Note: This plasmid is part of the Mendenhall and Meyers CRISPR-based Tagging system to add a tag (currently using FLAG) to endogenous proteins. Please see https://www.addgene.org/crispr/tagging/ for more details.
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.
For your Materials & Methods section:pFETCh_HMG20A was a gift from Eric Mendenhall & Richard M. Myers (Addgene plasmid # 86271 ; http://n2t.net/addgene:86271 ; RRID:Addgene_86271)
For your References section:CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts BS, Mackiewicz M, Mendenhall EM, Myers RM. Genome Res. 2015 Sep 9. 10.1101/gr.193540.115 PubMed 26355004