pDZ251 (pKAN 24xPP7SL cyc)
(Plasmid #35192)

• Depositing Lab: Robert Singer
• Publication: Larson et al Science. 2011 Apr 22;332(6028):475-8.

Backbone

• Vector backbone: pKAN
• Backbone manufacturer: David Miklos & Greg Freyer (Cold Spring Harbor Laboratory)
• Backbone size w/o insert (bp): 4194

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: 24xPP7SL cyc
• Species: Synthetic

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: M13rev
• 3′ sequencing primer: M13F20

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest for 35192

Terms and Licenses

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

Depositor Comments

This plasmid was designed to construct mRNAs that contain multiple copies of an RNA sequence (the 24xPP7 stem-loops) that can be recognized by a chimeric RNA-binding protein fused to GFP, encoded in plasmid pDZ276 (Addgene plasmid #35194). See associated publication for detailed methods and use of this plasmid.

The 24xPP7 stem loop cassettes in this plasmid were generated from two non-identical stem-loops to reduce unnecessary redundancy and the chance of recombination in bacteria. The sequence for a single cassette (with the two non-identical stem-loops in uppercase) is:

taaggtacctaattgcctagaaaGGAGCAGACGATATGGCGTCGCTCCctgcaggtcgactctagaaa
CCAGCAGAGCATATGGGCTCGCTGGctgcagtattcccgggttcatt