LV-Sox10MCS5-GFP-EF1a-mCherry
(Plasmid #115782)

• Purpose: Sox10-MCS5-GFP and a constitutive EF1α-mCherry reporter plasmid. Construct generates constitutive red fluorescence and basal promoter cfos conjugated SOXMCS5 enhancer driven GFP expression in a cell.
• Depositing Lab: Fraser Sim
• Publication: Pol et al Exp Neurol. 2013 Sep;247:694-702. doi: 10.1016/j.expneurol.2013.03.010. Epub 2013 Mar 16.

Backbone

• Vector backbone: pTRIP
• Backbone size w/o insert (bp): 9791
• Total vector size (bp): 2503
• Modifications to backbone: First, Sox10 multiple species conserved enhancer element 5 (MCS5, mouse CH15:79029307 CH15:79028761) (as described in Antonellis, et al., 2008 ) coupled with a c-fos basal promoter and eGFP was PCR amplified. This was inserted into a TOPO cloning vector and its sequence was verified. Then this insert was cloned into pTRIP-EF1α-mCherry (gift of Abdel Benraiss, University of Rochester) at the unique Spe1 restriction site.
• Vector type: Lentiviral

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mouse derived SOX10 Multiple Species Conserved enhancer element 5 conjugated with cfos basal promoter
• Species: Synthetic
• Insert Size (bp): 532
• Promoter: SOX10 Multiple Species Conserved enhancer element 5 coupled with cfos basal promoter
• Tag / Fusion Protein:
  • cfos promoter conjufated MCS5 enhancer fused to eGFP (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: TOPO Cloning
• 5′ sequencing primer: AAAACTAGTctgggtccctccccacccag
• 3′ sequencing primer: AAAACTAGTttacttgtacagctcgtccatgcc

Gene/Insert 2

• Gene/Insert name: EF1alpha promoter driven constitutive mCherry
• Species: Synthetic
• Insert Size (bp): 720
• Tag / Fusion Protein:
  • EF1α promoter fused to mCherry (N terminal on insert)

Resource Information

• Supplemental Documents:
  • Dual Sox10 - ReporterMCS5 sox10 -Ef1Alphamcherry.gb
• A portion of this plasmid was derived from a plasmid made by: The pXIG-Sox10-MCS5 plasmid contains Sox10 multiple species conserved enhancer element 5 (MCS5, mouse CH15:79029307CH15:79028761) coupled with a c-fos basal promoter and eGFP (as described in Antonellis, et al., 2008) was provided by A. McCallion of Johns Hopkins University. pTRIP-EF1α-mCherry lentivirus was provided by Dr. Pierre Charneau of the Pasteur Institute).

Terms and Licenses

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