pcDNA3.1/V5-His-TN-DROSHA
(Plasmid #106252)

• Purpose: Express TN-DROSHA in mammalian cells
• Depositing Lab: Joshua Mendell
• Depositing Organization: University of Texas Southwestern Medical Center
  Located in United States of America
• Publication: Chang et al Genome Res. 2015 Sep;25(9):1401-9. doi: 10.1101/gr.193607.115.

Backbone

• Vector backbone: pcDNA3/V5-His
• Backbone manufacturer: invitrogen
• Backbone size w/o insert (bp): 5514
• Total vector size (bp): 9533
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Drosha Ribonuclease III
• Alt name: Drosha
• Species: H. sapiens (human)
• Insert Size (bp): 4017
• Mutation: E1045Q and E1222Q mutations
• GenBank ID: NM_001100412
• Entrez Gene: DROSHA (a.k.a. ETOHI2, HSA242976, RANSE3L, RN3, RNASE3L, RNASEN)
• Promoter: CMV
• Tag / Fusion Protein:
  • V5-His (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pcDNA4/TO/cmycDrosha (Thomas Tuschl)

Terms and Licenses

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

Depositor Comments

Please note that DROSHA mutation numbering (E1045Q and E1222Q) is modeled on Heo et al., Mol Cell 32: 276–284. These mutations correspond to E1008Q and E1185Q in this construct. In addition, a series of silent mutations across bp 1203 - 1221render the encoded DROSHA mRNA resistant to siRNAs that target that site.

DNA (Catalog # 106252-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 106252-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.