pACT1:Cas9-GFP, U6:sgUPRT
(Plasmid #122853)

• Purpose: Expresses Cas9 fused with GFP and a sgRNA targeting the Cryptosporidium parvum UPRT gene
• Depositing Lab: David Sibley
• Publication: Wilke et al Cell Host Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi: 10.1016/j.chom.2019.05.007.

Backbone

• Vector backbone: pUC19
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 2700
• Total vector size (bp): 9981
• Vector type: CRISPR ; Cas9-GFP plasmid for genome editing of Cryptosporidium parasites

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: Cas9-GFP
• Species: streptococcus pyogenes
• Insert Size (bp): 4956
• Promoter: Cryptosporidium parvum actin
• Tag / Fusion Protein:
  • eGFP (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CCCAACACTTAACCTTTCAGT
• 3′ sequencing primer: TCAGTTCTATTCTCATTTGAAAGCT

Gene/Insert 2

• Gene/Insert name: U6-sgUPRT
• Species: Cryptosporidium parvum
• Insert Size (bp): 716
• Promoter: Cryptosporidium parvum U6

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATTTAGGCGCCAAGACCCAG
• 3′ sequencing primer: CTATAGTGTCACCTAAATAGCTTGGC

Resource Information

• Supplemental Documents:
  • pACT1-Cas9-GFP-U6-sgUPRT.gb

Terms and Licenses

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