pAf-CRISPR-yA
(Plasmid #191015)

• Purpose: CRISPR vector based on Aspergillus flavus U6 promoter and terminator for targeting the yA gene, which contains AMA1(the HindIII-PstI fragment) and ptrA selection marker.
• Depositing Lab: Perng-Kuang Chang
• Publication: Chang et al Microbiol Spectr. 2023 Feb 14;11(1):e0464822. doi: 10.1128/spectrum.04648-22. Epub 2023 Jan 18.

Backbone

• Vector backbone: pPTRII
• Backbone manufacturer: TaKaRa
• Backbone size w/o insert (bp): 10031
• Total vector size (bp): 13436
• Modifications to backbone: Removal of the PstI fragment of pPTRII (AMA1), 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: yA
• gRNA/shRNA sequence: TTAGTGAGAATCATTTGGCG
• 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_P4Y_3_yA_temp_pLann.gbk
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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