BCJJ125
(Plasmid #117515)

• Purpose: Cas9_3 (with intron) Golden Gate Level 0
• Depositing Lab: Jonathan D Jones
• Publication: Castel et al PLoS One. 2019 Jan 9;14(1):e0204778. doi: 10.1371/journal.pone.0204778. eCollection 2019.

Backbone

• Vector backbone: pICH41308
• Backbone size w/o insert (bp): 2247
• Total vector size (bp): 6708
• Vector type: Plant Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: BsaI:AATG:Cas9_3(intron):GCTT:BsaI
• Species: Synthetic
• Insert Size (bp): 4459
• Tags / Fusion Proteins:
  • 2xFLAG (N terminal on insert)
  • NLS (N terminal on insert)
  • NLS (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (not destroyed)
• 3′ cloning site: BsaI (not destroyed)
• 5′ sequencing primer: CGTTATCCCCTGATTCTGTGGATAAC
• 3′ sequencing primer: GTCTCATGAGCGGATACATATTTGAATG

Resource Information

• Supplemental Documents:
  • BCJJ125_Cas9-3.gbk
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Cloned by Baptiste Castel
This plasmid is unstable in E. coli. After transformation, many positive colonies will actually contains a truncated version of the insert, generally around the intron. The plasmid should be carefully verified on gel after restriction enzyme digestion to make sure the all insert is present. Please visit https://www.biorxiv.org/content/early/2018/09/17/419952 for bioRxiv preprint.