5'UTR CGG 99x FMR1-EGFP
(Plasmid #63091)

• Purpose: Contains expanded CGG repeats (99 repeats) within the 5'UTR of FMR1 as found in the Fragile X Tremor Ataxia Syndrome (FXTAS). These repeats are in frame with EGFP.
• Depositing Lab: Nicolas Charlet-Berguerand
• Publication: Charlet-Berguerand plasmids (unpublished)

Backbone

• Vector backbone: unknown
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 50 μg/mL
• Growth Temperature: Room Temperature
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: expanded CGG repeats (99 repeats) within the 5'UTR of FMR1
• Species: H. sapiens (human)
• Entrez Gene: FMR1 (a.k.a. FMRP, FRAXA, POF, POF1)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: CMV-F

Resource Information

• Supplemental Documents:
  • 5'UTR CGG99x FMR1 EGFP annotated genbank file
• Addgene Notes:
  • Addgene Diagnostic Test
  • Addgene Diagnostic Digest to verify insert- HindIII/XhoI
• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

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

Depositor Comments

Should be amplified in STBL3 bacteria and grown at Room temperature to keep the repeats stable.
This construct allows translation of CGG repeats into 99 glycine due to a near cognate ATG codon upstream of the repeat in the 5'UTR of FMR1 (an ACG codon that initiates weakly translation). These CGG are in frame with the EGFP tag.

This plasmid should be verified via restriction analysis. Multiple colonies (at least 6) may need to be screened to isolate the correct construct. The plasmid should be screened for size every time it is amplified. We suggest a digest with HindIII and XhoI and running a 2% agarose gel. If correct expect 2 bands: 4218bp and 470bp. Smaller band is the insert.