Cas12aUltra_5XNLS
(Plasmid #218775)

• Purpose: His-TEV-Cas12aUltra-BPSV40_cMyc_NP_5NLS
• Depositing Lab: Ross Wilson
• Publication: Sahu et al bioRxiv 2024.07.14.603391

Backbone

• Vector backbone: 3xNLS-NLP-cMyc-cMyc AspCas12a
• Backbone size w/o insert (bp): 5443
• Total vector size (bp): 9626
• Modifications to backbone: Addition of bipartite SV40 nuclear localization signals (NLS) at the -N and -C termini of Cas12 expressing gene, change in c-myc NLS sequence at the C-terminus of the Cas12 expressing gene
• Vector type: Bacterial Expression, CRISPR

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: 5xNLS-BPSV40-NLP-cMyc-cMyc-BPSV40- AspCas12a
• Alt name: Cas12a ultra 5NLS
• Species: Acidaminococcus
• Insert Size (bp): 4287
• Mutation: Nucleotides mutations were made to encode for arginine at position 537 and leucine at position 870 of the Cas12 protein
• Tags / Fusion Proteins:
  • 6XHis (C terminal) (C terminal on insert)
  • HA (C terminal on insert)
  • SV40 (C terminal on insert)
  • Nucleoplasmin (C terminal on insert)
  • c-myc NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • Cas12a ultra 5NLS.gbk
• A portion of this plasmid was derived from a plasmid made by: Plasmid 3xNLS-NLP-cMyc-cMyc AspCas12a was purchased from Addgene (plasmid #182122 from Scot Wolfe, RRID:Addgene_182122)

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.1101/2024.07.14.603391 for bioRxiv preprint.