pSox2-bd::FP
(Plasmid #34703)

• Depositing Lab: Hollis Cline
• Publication: Bestman et al J Comp Neurol. 2012 Feb 1;520(2):401-33. doi: 10.1002/cne.22795.

Backbone

• Vector backbone: act-GVP-UG
• Backbone manufacturer: Created by Köster and Fraser
• Vector type: Xenopus expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: eGFP
• Tags / Fusion Proteins:
  • 6x Sox2/Oct3-4 transcription factor binding domain (N terminal on insert)
  • 165-bp minimal FGF4 promoter (N terminal on insert)
  • Gal4-UAS (N terminal on insert)
  • 165-bp minimal FGF4 promoter (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: none (unknown if destroyed)
• 3′ cloning site: none (unknown if destroyed)
• 5′ sequencing primer: pBluescriptSK_primer
• 3′ sequencing primer: EGFP_N

Resource Information

• Supplemental Documents:
  • p095 6xSO Gal-UAS mFGF eGFP.gb
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

The Sox2/Oct3-4 enhancer elements and a minimal promoter from the murine FGF4 were subcloned from the plasmid described in Figure 1 of Ambrosetti et al. (2000). The fragment contains six tandem repeats of the 35-bp fragment containing the Sox2/Oct3-4 heterodimer transcription factor binding domain (bd) followed by a 165-bp minimal FGF4 promoter (mFGF4). This plasmid was a gift of Claudio Basilico (New York University). To increase the FP signal, the Sox2/Oct 3-4-mFGF regulatory fragment was moved to the “act-GVP-UG” concatenated plasmid created by Köster and Fraser (2001), which was adapted from the Drosophila Gal4-UAS system (Brand and Perrimon,1993) and shown to function in Xenopus (Chae et al.,2002; Hartley et al.,2002). We replaced the minimal promoters of the act-GVP-UG plasmid with our Sox2/Oct3-mFGF regulatory fragment to control the expression of the transcriptional activator, Gal4-VP16, which in turn drives the expression of genes controlled by the upstream activating sequence (UAS).

The final expression vector (abbreviated as pSox2-bd::FP) contains: 6 repeats of the Sox2/Oct3-4 transcription factor binding domain, mFGF4, Gal4-VP16, a polyadenylation site, 14 repeats of UAS, mFGF4, and eGFP (ClonTech, Palo Alto, CA).

Sequencing with EGFP-N identified 1nt deletion at position 1733, 1 nt insertion at position 1214, 2 nt insertion at 1557 compared to the depositor provided sequence. These mutations are all in non-coding regions and have been shown by the depositing lab to not effect the function of the plasmid.