Addgene - pDZ251 (pKAN 24xPP7SL cyc) Plasmid Data

Plasmid 35192: pDZ251 (pKAN 24xPP7SL cyc)

Gene/insert name:
24xPP7SL cyc
Species:
Synthetic
Vector backbone:
pKAN
(Search Vector Database)
Backbone manufacturer:
David Miklos & Greg Freyer (Cold Spring Harbor Laboratory)
Vector type:
Synthetic
Backbone size w/o insert (bp):
4194
Cloning site 5':
unknown
Site destroyed during cloning:
Unknown
Cloning site 3':
unknown
Site destroyed during cloning:
Unknown
5' sequencing primer:
M13rev List of Sequencing Primers
3' sequencing primer:
M13F20
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
Unknown
Sequence: View sequences (5)
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Robert Singer
Terms and Licenses:
MTA

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

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Real-time observation of transcription initiation and elongation on an endogenous yeast gene. Larson et al (Science. 2011 Apr 22;332(6028):475-8. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 35192" in your Materials and Methods section.