pSico-CAG-v-H-Ras-IRES-Luciferase/EGFP
(Plasmid #58959)

• Purpose: lentiviral expression of v-H-Ras and luciferase/EGFP reporter
• Depositing Lab: Snorri Thorgeirsson
• Publication: Holczbauer et al Gastroenterology. 2013 Jul;145(1):221-31. doi: 10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19.

Backbone

• Vector backbone: pSICO
• Backbone size w/o insert (bp): 5800
• Total vector size (bp): 11112
• Modifications to backbone: The loxP sites were deleted and the vector was modified for use with the Gateway Cloning System
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: EGFP

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: CMV enhancer/chicken beta actin promoter
• Insert Size (bp): 1605

Cloning Information for Gene/Insert 1

• Cloning method: Gateway Cloning
• 5′ sequencing primer: none
• 3′ sequencing primer: IRES-R

Gene/Insert 2

• Gene/Insert name: v-H-Ras
• Alt name: Harvey murine sarcoma virus p21 transforming protein
• Insert Size (bp): 570
• GenBank ID: M24154
• Promoter: CMV enhancer/chicken beta actin

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: none
• 3′ sequencing primer: IRES-R

Gene/Insert 3

• Gene/Insert name: luc2/EGFP fusion gene
• Alt name: luc2 synthetic firefly luciferase/enhanced green fluorescent protein gene
• Species: Synthetic
• Insert Size (bp): 2391
• Promoter: CMV enhancer/chicken beta actin

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: IRES-F
• 3′ sequencing primer: EGFP-C

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This vector was constructed by the Protein Expression Laboratory at the Frederick National Laboratory for Cancer Research. The v-H-Ras gene was obtained from plasmid pBW1423 (Willumsen BM et al. The bovine papillomavirus E5 oncogene can cooperate with ras: identification of p21 amino acids critical for transformation by c-rasH but not v-rasH. Mol Cell Biol. 1991 Dec;11(12):6026-6033).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is a modified version of the plasmid used by Holczbauer et al. The SV40 enhancer/FerH promoter in the original vector has been replaced by the CMV enhancer/chicken beta actin promoter.