B52_puro_gRNA_3418711_KCNB2
(Plasmid #197558)

• Purpose: gRNA targeting upstream region of KCNB2 (site 3418711, control for epigenetic targeting)
• Depositing Lab: Darius Bägli
• Publication: Persistent changes in KCNB2 expression and CpG methylation is associated with ongoing hypertrophic changes after release of partial outlet obstruction. (unpublished)

Backbone

• Vector backbone: CMV+Puro_B52
• Backbone manufacturer: Bagli Lab
• Backbone size w/o insert (bp): 4044
• Total vector size (bp): 4778
• Modifications to backbone: addition of gRNA to region 3418711 upstream of KCNB2
• Vector type: Mammalian Expression ; gRNA with puromycin selection
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: upstream KCNB2 promoter gRNA
• Alt name: upstream Kv2.2 promoter gRNA at chr5:3418711 site
• Species: R. norvegicus (rat)
• Insert Size (bp): 734
• GenBank ID: NM_054000
• Entrez Gene: Kcnb2 (a.k.a. Kv2.2)
• Promoter: U6
• Tag / Fusion Protein:
  • puromycin resistance cassette under CMV promotion

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: waiting to find
• 3′ sequencing primer: waiting to find

Resource Information

• A portion of this plasmid was derived from a plasmid made by: We had help from Genscript with the cloning, though the design was ours.

Terms and Licenses

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

Depositor Comments

Any publications using this clone should state: "we thank the laboratory of Darius Bagli for the use of this clone". In addition, we have a forthcoming article which can be referenced once it is published.