hVoS 1.5 thy-1
(Plasmid #45281)

• Depositing Lab: Meyer B. Jackson
• Publication: Wang et al Biophys J. 2010 Oct 6;99(7):2355-65. doi: 10.1016/j.bpj.2010.07.037.

Backbone

• Vector backbone: pUC18
• Backbone size w/o insert (bp): 2686
• Modifications to backbone: pUC18 modification (see Feng et al. 2010): The thy1 vector contains 6.5 kb of the murine thy1.2 gene, extending from the promoter to the intron following exon 4, but lacking exon 3 and its flanking introns. The deleted sequences are required for expression in nonneural cells but not in neurons, whereas sequences in upstream introns are required for expression in neurons. Reporters are cloned into an XhoI site, effectively replacing the deleted sequences.
• Vector type: Targeting

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): JM109
• Growth instructions: This plasmid has only been grown on Carbenicillin and has not be verified to grow well on Amp plates.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: CeFP
• Alt name: cerulean fluorescent Protein
• Promoter: thy1
• Tag / Fusion Protein:
  • truncated h-ras motif (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (destroyed during cloning)
• 3′ cloning site: XhoI (destroyed during cloning)
• 5′ sequencing primer: eGFP-N
• 3′ sequencing primer: eGFP-C

Terms and Licenses

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

Depositor Comments

This construct was made to create transgenic mice in which hVoS 1.5 is expressed under the thy1 promoter only in neuronal cells.

The cerulean sequence has a silent G-C mutation at 687 that differs from standard sequence.

Addgene has sequenced a portion of this plasmid which confirms part of thy-1 with some ambiguous bases in the sequencing results.