pSBbi-Bla-dCas9-KRAB-MeCP2_hU6-SapI
(Plasmid #196074)

• Purpose: (Empty Backbone) Constitutive CRISPRi vector conferring blasticidin S resistance, ready for insertion of gRNA targeting gene of interest using SapI restriction sites.
• Depositing Lab: Joanna Bereta
• Depositing Organization: Jagiellonian University
  Located in Poland
• Publication: Czarnek et al Mol Biotechnol. 2023 Jan 28. doi: 10.1007/s12033-023-00675-5.

Backbone

• Vector backbone: pSBbi-Bla
• Backbone manufacturer: Kowarz et al., 2015
• Backbone size w/o insert (bp): 5656
• Total vector size (bp): 11139
• Vector type: Mammalian Expression
• Selectable markers: Blasticidin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: dCas9-KRAB-MeCP2 repressor
• Species: Streptococcus pyogenes
• Insert Size (bp): 5313
• Mutation: Catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system (Cas9m4) fused with Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF1 (Margolin et al., 1994) and transcriptional repression domain from rat methyl-CpG-binding protein 2 (Nan et al., 1997).

Gene/Insert 2

• Gene/Insert name: hU6 promoter and gRNA scaffold
• Species: H. sapiens (human); Streptococcus pyogenes
• Insert Size (bp): 343
• Mutation: hU6 promoter and gRNA scaffold containing SapI sites were amplified from pX330 containing SapI sites instead of BbsI sites (Czarnek et al., 2021).

Terms and Licenses

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

DNA (Catalog # 196074-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 # 196074-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.