PurposeVector to measure enhancer responsiveness of candidates from a genomic library (cloned instead of the core promoter) by determining the abundance of transcripts originating from each candidate.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||86386||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3426
Vector typeSTAP-seq screening vector
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth Strain(s)ccdB Survival
Copy numberHigh Copy
Gene/Insert nameD. melanogaster tj enhancer
SpeciesD. melanogaster (fly)
Insert Size (bp)856
- Cloning method Restriction Enzyme
- 5′ cloning site unknown (unknown if destroyed)
- 3′ cloning site unknown (unknown if destroyed)
- 5′ sequencing primer RVprimer3 (Common Sequencing Primers)
to consruct the STAP-seq screening vector from pGL3 Promoter we replaced the sequence between the BglII and FseI sites with the following sequence, containing a ccdB suicide gene next to the Chloramphenicole resistance gene flanked by homology arms (used for cloning the candidates during library generation), an intron (mhc16), an ORF (truncated sgGFP, Qbiogene, Inc)
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pSTAP-seq_fly-tj was a gift from Alexander Stark (Addgene plasmid # 86386 ; http://n2t.net/addgene:86386 ; RRID:Addgene_86386)
For your References section:Genome-wide assessment of sequence-intrinsic enhancer responsiveness at single-base-pair resolution. Arnold CD, Zabidi MA, Pagani M, Rath M, Schernhuber K, Kazmar T, Stark A. Nat Biotechnol. 2016 Dec 26. doi: 10.1038/nbt.3739. 10.1038/nbt.3739 PubMed 28024147