pAf-CRISPR-ctfR1
(Plasmid #207992)

• Purpose: CRISPR vector used with pAf-CRISPR-phoA (#207991) to delete both aflatoxin and cyclopiazonic acid gene clusters of Aspergillus flavus
• Depositing Lab: Perng-Kuang Chang
• Publication: Chang et al Fungal Genet Biol. 2023 Dec 27;170:103863. doi: 10.1016/j.fgb.2023.103863.

Backbone

• Vector backbone: pPTRII
• Backbone manufacturer: TaKaRa
• Backbone size w/o insert (bp): 10031
• Total vector size (bp): 13733
• Modifications to backbone: Removal of non-functional sequence of AMA1 in pPTRII, expression of cas9 using Aspergillus nidulans gpdA promoter and trpC terminator, expression of sgRNA using Aspergillus flavus U6 promoter and terminator
• Vector type: 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: ctfR1
• gRNA/shRNA sequence: GCTCTGAGCTGGCATTCCGC
• Species: Aspergillus flavus
• Promoter: Aspergillus flavus U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PstI (unknown if destroyed)
• 3′ cloning site: KpnI (unknown if destroyed)
• 5′ sequencing primer: ATACTGCAGTTCTCTTTAGAATTCAACTGTGGGT
• 3′ sequencing primer: TATGGTACCACATATTTAAAAAAAGTCTCCTGCC

Resource Information

• Supplemental Documents:
  • Chang_V9Y_2_CRISPR-ctfR1_pLann.gbk

Terms and Licenses

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