JDW 770 (pTol2-kdrl:EGFP-hsKRAS4B-G12V-ac/Y)
(Plasmid #156414)

• Purpose: kdrl promoter driving EGFP fused to human KRAS4B-G12V with an alpha-crystallin Venus (YFP) reporter flanked by Tol2 sites
• Depositing Lab: Joshua Wythe
• Publication: Fish et al Circ Res. 2020 Jun 17. doi: 10.1161/CIRCRESAHA.119.316500.

Backbone

• Vector backbone: pDEST_Tol2
• Vector type: Zebrafish Transgenesis

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: human mutant KRAS4B-G12V
• Species: H. sapiens (human)
• Insert Size (bp): 1314
• Mutation: mutant KRAS4B-G12V
• GenBank ID: NM_004985.5
• Entrez Gene: KRAS (a.k.a. 'C-K-RAS, C-K-RAS, CFC2, K-RAS2A, K-RAS2B, K-RAS4A, K-RAS4B, K-Ras, K-Ras 2, KI-RAS, KRAS1, KRAS2, NS, NS3, OES, RALD, RASK2, c-Ki-ras, c-Ki-ras2)
• Promoter: kdrl
• Tag / Fusion Protein:
  • EGFP (N terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: V5 REV: 5'-CGTAGAATCGAGACCGAGG; V5 FWD: 5'-GGTAAGCCTATCCCTAACCC
• 3′ sequencing primer: KRAS REV2: ctgtattgtcggatctccctc; KRAS FWD2: 5'-gagggagatccgacaatacag;

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry