Tol2 Krt4 Src (YF)-uniRapR-cerulean
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||45509||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepT2KHspGGFF (Asakawa K, Kawakami K. Dev Growth Differ. 2008)
- Backbone size w/o insert (bp) 9773
- Total vector size (bp) 13668
Modifications to backboneT2:Krt4: Src-iFKB-cpFRB-Cer-Myc was constructed by first removing the heat shock promoter from pT2KHspGGFF (Asakawa K, Kawakami K. Dev Growth Differ. 2008) via a ApaI/NcoI digest and replacing it with a ApaI/NcoI fragment containing the Krt4 promoter (Ju B et al. Dev Genet. 1999, Gong et al. 2002. Dev Dyn, genebank GenBank: AF440690.1) which was amplified with (Apa-Krt4 GATGGGCCCCTACAGTAAAGCTTCTCCACAATGTCCCG and NcoI-Krt4 CTACCATGGCTCTGCGTGTCTCTCAGCAGC). Next the GFP-Gal4 fragment was removed with a NcoI/SmaI digest and replaced with the cassette containing Src-iFKB-cpFRB-Cer-Myc which had been removed from PUse via a NcoI/SnaI digest. The final T2:Krt4: Src-iFKB-cpFRB-Cer-Myc contains the Krt4 promoter, the Src-iFKB-cpFRB-Cer-Myc cassette and the polyA signal between Tol2 sequences.
Vector typezebrafish expression
Growth in Bacteria
Gene/Insert nameSrc (YF)-uniRapR-cerulean
Insert Size (bp)2988
- Promoter krt4
/ Fusion Protein
- cerulean (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NcoI (not destroyed)
- 3′ cloning site Snab1 (destroyed during cloning)
- 5′ sequencing primer CAAGGGTTGCCATCAAAACT
- 3′ sequencing primer AAGTCGTGCTGCTTCATGTG (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
Please note that Addgene's sequencing results differ from the full plasmid sequence information from the depositing laboratory at bp# 3150, 3188, 3193, 3199, 3211, 3227, 3307, 5577 & 5578. These differences are not a concern for the function of the plasmid according to the depositing laboratory.
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:Tol2 Krt4 Src (YF)-uniRapR-cerulean was a gift from Klaus Hahn & Anna Huttenlocher (Addgene plasmid # 45509 ; http://n2t.net/addgene:45509 ; RRID:Addgene_45509)
For your References section:Rational design of a ligand-controlled protein conformational switch. Dagliyan O, Shirvanyants D, Karginov AV, Ding F, Fee L, Chandrasekaran SN, Freisinger CM, Smolen GA, Huttenlocher A, Hahn KM, Dokholyan NV. Proc Natl Acad Sci U S A. 2013 Apr 23;110(17):6800-4. doi: 10.1073/pnas.1218319110. Epub 2013 Apr 8. 10.1073/pnas.1218319110 PubMed 23569285