p-dCas9-LSD1-K661A-Hygro
(Plasmid #104414)

• Purpose: transient expression of dCas9-dLSD1 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• Publication: Beck lab Cas9 plasmids (unpublished)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: dLSD1
• Species: H. sapiens (human)
• Mutation: IRATp970A0364D (Source Bioscience) has a P405H change that was reverted to the consensus. K661A
• Entrez Gene: KDM1A (a.k.a. AOF2, BHC110, CPRF, KDM1, LSD1)
• 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). LSD1A isoform B catalytic domain (a.a. 171-852) was from LSD1 cDNA clone IRATp970A0364D (Source Bioscience). The P405H change in this cDNA clone was reverted to the consensus during Gibson assembly. K661A mutation was introduced by PCR.