pT3TS nErCas12an
(Plasmid #132642)

• Purpose: Used for in vitro transcription of dual NLS ErCas12a mRNA
• Depositing Lab: Stephen Ekker
• Publication: Wierson et al CRISPR J. 2019 Dec;2(6):417-433. doi: 10.1089/crispr.2019.0026. Epub 2019 Nov 19.

Backbone

• Vector backbone: pT3TS
• Backbone manufacturer: Stephen Ekker
• Backbone size w/o insert (bp): 3216
• Total vector size (bp): 6994
• Vector type: CRISPR ; In Vitro transcription

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: ErCas12a
• Alt name: Mad7
• Species: Eubacterium rectale
• Insert Size (bp): 3843
• Promoter: T3
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: SacII (not destroyed)
• 5′ sequencing primer: aattaaccctcactaaagg
• 3′ sequencing primer: taatacgactcactatagg

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This gene was ordered as a GeneBlock from Jeff Essner's laboratory

Terms and Licenses

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

Depositor Comments

Original gene sequence was obtained from Inscripta and zebrafish codon optimized and ordered as a gene block.