pSLQ2827 pHR: U6-SpsgSV40 CMV-PYL1-KRAB-IRES-mCherry
(Plasmid #84260)

• Purpose: Expresses Sp sgSV40 gRNA with ABA-inducible KRAB and mCherry for diametric regulation
• Depositing Lab: Stanley Qi
• Publication: Gao et al Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.

Backbone

• Vector backbone: pHR
• Total vector size (bp): 9086
• Vector type: Mammalian Expression, Lentiviral, CRISPR

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Sp sgSV40
• Alt name: sgRNA sequence: GAAAGTCCCCAGGCTCCCCAGC
• Species: Synthetic; S. pyogenes
• Promoter: mouse U6

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BstXI (destroyed during cloning)
• 3′ cloning site: XhoI (destroyed during cloning)
• 5′ sequencing primer: GAGATCCAGTTTGGTTAGTACCGGG
• 3′ sequencing primer: ATGCATGGCGGTAATACGGTTAT

Gene/Insert 2

• Gene/Insert name: PYL1-KRAB
• Species: A. thaliana (mustard weed), Synthetic
• Insert Size (bp): 804
• Promoter: CMV
• Tags / Fusion Proteins:
  • PYL1 (N terminal on insert)
  • IRES-mCherry (C terminal on backbone)

Cloning Information for Gene/Insert 2

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: gctaccatgggtgggggcgcgc
• 3′ sequencing primer: gacggcaatatggtggaaaataac

Resource Information

• Supplemental Documents:
  • pSLQ2827 Annotated GenBank File
• A portion of this plasmid was derived from a plasmid made by: The PYL1 domain was a gift from Jerry Crabtree (Stanford).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry