eSpCas9(1.1)_NoFlag_ATP1A1_G3_SLBP_G1_Dual_sgRNA
(Plasmid #191528)

• Purpose: Vector for tandem expression of SLBP 3'UTR G1 sgRNA in combination with ATP1A1 G3 sgRNA from two independent U6 promoters to facilitate SLBP endogenous tagging by marker free coselection using ouabain
• Depositing Lab: Yannick Doyon
• Publication: S-phase specific probe (unpublished)

Backbone

• Vector backbone: eSpCas9(1.1)_No_FLAG_ATP1A1_G3_Dual_sgRNA (Plasmid #86613)
• Backbone size w/o insert (bp): 8858
• Total vector size (bp): 8860
• Vector type: Mammalian Expression, CRISPR ; Co-selection via HDR using ouabain

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: SLBP 3'UTR G1 sgRNA + ATP1A1 G3 sgRNA + FLAGless enhanced specificity Cas9 (1.1)
• Species: H. sapiens (human)
• Insert Size (bp): 20
• GenBank ID:

Resource Information

• Supplemental Documents:
  • eSpCas9 (1.1)_NoF_Dual_G1_SLBP_G3_ATP1A1.gb

Terms and Licenses

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

Depositor Comments

-ATP1A1 G3 gRNA target sequence is GAGTTCTGTAATTCAGCATA

-SLBP G1 gRNA target sequence is GTGAGCTAACTGCCCCCTGG (1st base "A" is mismatched/replaced by "G").

-Use in combination with hSLBP_mNG_FS_Donor (Plasmid #191387)

-See Marker-free coselection for CRISPR-driven genome editing in human cells. Agudelo D, Duringer A, Bozoyan L, Huard CC, Carter S, Loehr J, Synodinou D, Drouin M, Salsman J, Dellaire G, Laganiere J, Doyon Y. Nat Methods. 2017 Apr 17. doi: 10.1038/nmeth.4265. 10.1038/nmeth.4265 PubMed 28417998