pTC201B
(Plasmid #52193)

• Purpose: for creating reversible targeting construct
• Depositing Lab: Tian Chi
• Publication: Chaiyachati et al BMC Biol. 2012 Nov 30;10:96. doi: 10.1186/1741-7007-10-96.

Backbone

• Vector backbone: pEZ-Frt-lox-DT
• Backbone manufacturer: Klaus Rajewsky (Addgene plasmid # 11736)
• Backbone size w/o insert (bp): 4500
• Modifications to backbone: The FRT-flanked cassette in pEZ was replaced by a synthetic fragment bearing an FRT-flanked splicing acceptor together with multiple cloning sites. A Neo-Ires-GFP-SV40 polyA fragment was inserted immediately downstream of the splicing acceptor to create the gene-trapping cassette. The left and right arms flanking the cassette, encompassing BAF57 exons 2 to 3 and exon 4, respectively, were amplified by PCR from C57B/6 mice
• Vector type: Cre/Lox
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl2
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: promoter-trap cassette Neo-Ires-GFP

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T3
• 3′ sequencing primer: SV40pA-R

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This plasmid carries the FRT-flanked promoter-Neo expression cassette, which is in turn flanked by LoxP site.

The targeted upstream trapped exon must be in frame with the Neo sequence in the vector in order to produce the exon-Neo fusion protein.