PurposeDonor vector for 3' FLAG tag of human MBD1_iso1
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||86263||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberHigh Copy
Gene/Insert nameMBD1_iso1 homology arms
SpeciesH. sapiens (human)
Entrez GeneMBD1 (a.k.a. CXXC3, PCM1, RFT)
/ 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
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Donor plasmid for tag insertion of MBD1 gene at position hg19 chr18:47797873.This plasmid was co-transfected along with PX458_MBD1_iso1_1 and PX458_MBD1_iso1_2 into HepG2 cells, and genomic editing was verified by PCR. 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 MBD1. 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_MBD1_iso1 was a gift from Eric Mendenhall & Richard M. Myers (Addgene plasmid # 86263 ; http://n2t.net/addgene:86263 ; RRID:Addgene_86263)
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