pEF004-sgRNA-shuffle-in
(Plasmid #104440)

• Purpose: Provide and shuffle a cassette of AtU6:sgRNA-transRNA into SM-destination vectors (pRW006 and pRW004) with golden gate cloning strategy. Work together with pEF005
• Depositing Lab: Detlef Weigel
• Publication: Wu et al Plant Methods. 2018 Aug 2;14:65. doi: 10.1186/s13007-018-0330-7. eCollection 2018.

Backbone

• Vector backbone: pD000
• Backbone manufacturer: Lampropoulos et al., 2013
• Total vector size (bp): 2934
• Modifications to backbone: A U6 promoter driven sgRNA-transRNA cassette is cloned into the backbone
• Vector type: Bacterial Expression, Plant Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: sgRNA-transRNA transcription by U6 promoter
• Alt name: gRNA-transRNA
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 234
• Promoter: U6 promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: M13 forward
• 3′ sequencing primer: M13 reverse

Terms and Licenses

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