LV-Sox10MCS5-GFP
(Plasmid #115783)

• Purpose: This plasmid encodes for Sox10-MCS5-GFP reporter. Cell carrying this construct expresses green fluorescence under the control of SOX-MCS5 enhancer conjugated with cfos basal promoter.
• 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: SIN18
• Backbone size w/o insert (bp): 6378
• Total vector size (bp): 7906
• Modifications to backbone: First, Sox10 multiple species conserved enhancer element 5 (MCS5, mouse CH15:79029307 CH15:79028761) coupled with a c-fos basal promoter was excised using Xho1/Age1 restriction enzyme digest from pXIG-MCS5Sox10-eGFP (plasmid described in Antonellis et al 2008). This was inserted into aMHC-eGFP-Res-Neo vector (Add gene # 21229) to replace the aMHC promoter. Rex-Neo selection cassette was removed by digesting the plasmid with Kpn1 and Spe1 plasmids, followed by blunting and blunt end ligation.
• 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

• Gene/Insert name: mouse derived SOX10 Multiple Species Conserved enhancer element 5 conjugated with cfos basal promoter
• Species: M. musculus (mouse)
• Insert Size (bp): 546
• Promoter: SOX10 Multiple Species Conserved enhancer element 5 coupled with cfos basal promoter
• Tag / Fusion Protein:
  • cfos basal promoter conjugated MCS5 promoter fused to eGFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xho1 (unknown if destroyed)
• 3′ cloning site: Age1 (unknown if destroyed)
• 5′ sequencing primer: AAAACTAGTctgggtccctccccacccag
• 3′ sequencing primer: AAAACTAGTttacttgtacagctcgtccatgcc

Resource Information

• Supplemental Documents:
  • Single SOX10_MCS5-eGFP reporter.gb
• Addgene Notes:
  • Addgene Diagnostic Digest
• 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. The single reporter lentivirus was generated by cloning Sox10 -MCS5:cfos enhancer/promoter cassette into the aMHCeGFP-Rex-Neo lentiviral reporter (AddGene, #21229) ( Kita-Matsuo et al., 2009 )

Terms and Licenses

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