pUC19 KGA HA5'-mKO2-P2A-Zeo-3'
(Plasmid #110408)

• Purpose: Homology donor for glutaminase KGA isoform knock-in, mKO2-P2A-Zeo
• Depositing Lab: Sandra Martha Gomes Dias
• Publication: HuR (ELAVL1) silencing sensitizes triple-negative breast cancer to glutaminase-targeted therapy (unpublished)

Backbone

• Vector backbone: pUC19
• Backbone manufacturer: #50005
• Backbone size w/o insert (bp): 2686
• Total vector size (bp): 5797
• Vector type: CRISPR
• Selectable markers: Puromycin

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: GLS glutaminase
• Species: H. sapiens (human)
• Insert Size (bp): 3132
• Entrez Gene: GLS (a.k.a. AAD20, CASGID, DEE71, EIEE71, GAC, GAM, GDPAG, GLS1, KGA)
• Promoter: none

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: M13 Reverse 5' CAGGAAACAGCTATGAC 3'

Resource Information

• Supplemental Documents:
  • pD092-pUC19 KGA HA5'-mKO2-P2A-Zeo-3'.gb

Terms and Licenses

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

Depositor Comments

Homology arm for C-Terminal knock-in at stop codon of GLS' KGA isoform. mKO2-P2A knock-in.

Construction order:
GLS 1kb 5' Homology Arm: SacI-BamHI;
GLS 1kb 3' Homology Arm BamHI-SalI;
BglII-BamHI mKO2
BglII-BamHI P2A
BglII-BamHI BleoR

System options:
sgRNA:
pX330.puro sgRNA KGA Stop1 (Addgene #110405)

Homology arms:
pUC19 KGA HA5'-mKO2-3' (Addgene #110406)
pUC19 KGA HA5'-mKO2-P2A-3' (Addgene #110407)
pUC19 KGA HA5'-mKO2-P2A-Zeo-3' (Addgene #110408)