pCasTet-λ
(Plasmid #128318)

• Purpose: Constitutive expression of cas9 and anhydrotetracycline/tetracycline inducible expression of lamda RED. Useful variant of Plasmid #62225 when arabinose induction is not possible.
• Depositing Lab: Christopher Johnston
• Publication: Johnston et al Proc Natl Acad Sci U S A. 2019 May 16. pii: 1820256116. doi: 10.1073/pnas.1820256116.

Backbone

• Vector backbone: Plasmid #62225
• Backbone manufacturer: addgene
• Modifications to backbone: Replacement of arabinose inducible lamda RED system with tetracycline/anhydrotetracycline inducible lamda RED system.
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Must be grown at 30C! Vector has temperature-sensitive replication (RepA101ts)
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: TetR and pTetR/TetO
• Species: E. coli
• Insert Size (bp): 818
• Mutation: Constitutive expression of cas9 and anhydro-tetracycline/tetracycline inducible expression of LambdaRED and sgR
• Promoter: tet promoter

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: caattgatcgtaaacgatatacgtcta
• 3′ sequencing primer: ctcaagacgatcctgaatgtaataa

Terms and Licenses

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