p-dCas9-dTET1-H1672Y-D1674A-Hygro
(Plasmid #104413)

• Purpose: transient expression of dCas9-dTET1 fusion protein
• Depositing Lab: Stephan Beck
• Publication: Beck lab Cas9 plasmids (unpublished)

Backbone

• Vector backbone: pMLM3705
• Backbone manufacturer: Keith Joung (Addgene plasmid # 47754)
• Backbone size w/o insert (bp): 9075
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: dTET1
• Species: H. sapiens (human)
• Mutation: H1672Y-D1674A
• Promoter: CMV promoter

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

Backbone modification: added Hygromycin resistance cassette (PGK promoter and Hygromycin resistance gene from Addgene Plasmid 41721 (MSCVhygro- F-G9a), SV40 early polyadenylation signal from the pCDNA3.1 backbone of Addgene Plasmid 13820 (HDAC1 Flag). TET1 catalytic domain (amino acids 1418- 2136) was from Addgene plasmid 39454 (pAAV-EF1a-HA-hTet1CD- WPRE-PolyA). The H1672Y-D1674A mutations were added by PCR.

Use of this plasmid is described in:
Baumann, V., Wiesbeck, M., Breunig, C. T., Braun, J. M., Köferle, A., Ninkovic, J., et al. (2019). Targeted removal of epigenetic barriers during transcriptional reprogramming. Nature Communications, 1–12. http://doi.org/10.1038/s41467-019-10146-8