pCMU-4
(Plasmid #172353)

• Purpose: CRISPR gene editing in Streptomyces, codon-optimized Cas9 behind theophylline riboswitch and D4 promoter, with custom gRNA
• Depositing Lab: Eriko Takano
• Publication: Ye et al J Ind Microbiol Biotechnol. 2020 May;47(4-5):413-423. doi: 10.1007/s10295-020-02277-5. Epub 2020 May 4.

Backbone

• Vector backbone: pCRISPomyces-2
• Backbone size w/o insert (bp): 10994
• Total vector size (bp): 10875
• Modifications to backbone: Removal of rpsL promoter for cas9. Replaced with D4, additional theophylline responsive riboswitch.
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Apramycin, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: D4-TheoR-cas9
• Alt name: Codon optimised cas9 behind D4 promoter and theophylline riboswitch
• Alt name: Cloned by Gibson Assembly
• Species: Synthetic
• Insert Size (bp): 4280
• GenBank ID:
• Promoter: D4

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ACGATCCTCGCCCTGCT
• 3′ sequencing primer: GACGATGTTGCCGAAGAT

Resource Information

• Supplemental Documents:
  • pCMU-4.gbk

Terms and Licenses

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