MultiMate-HITI-2c ACTB
(Plasmid #206267)

• Purpose: All in one recombinant baculovirus production vector. Encodes Cas9, sgRNA and donor for homology independent targeted integration in the ACTB locus.
• Depositing Lab: Imre Berger
• Publication: Aulicino et al Nucleic Acids Res. 2022 Jul 22;50(13):7783-7799. doi: 10.1093/nar/gkac587.

Backbone

• Vector backbone: pMMACE DEST CMV SpCas9
• Vector type: Mammalian Expression, CRISPR ; Recombinant baculovirus production
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Gentamicin, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SpCas9 (S. Pyogenes), hACTB HITI-2c donor, mCherry, Puro-R, hU6 hACTB sgRNA, AcrIIA4, eGFP
• Species: H. sapiens (human)

Cloning Information

• Cloning method: Gateway Cloning

Resource Information

• Supplemental Documents:
  • MultiMate-HITI-2c ACTB.gb
• A portion of this plasmid was derived from a plasmid made by: SpCas9 was PCR amplified from px459 (Addgene plasmid #48139, Ran et al. Nat Protocol 2013)sgRNAs cloning was performed by overlap extent PCRs of hU6 and scaffold fragments amplified from px459 (Addgene plasmid#48139, Ran et al. Nat Protocol 2013)HDR and HITI-2c templates were amplified by Gibson assembly of genomic DNA fragments, mCherry was amplified from (Addgene plasmid#24304) T2A Puro from px459 (Addgene plasmid#48139, Ran et al. Nat Protocol 2013)polH/J23119 AcrII4 (Rauch B.J. et al. Cell 2017) was synthesised by Twist Bioscience.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry