PurposepSIN - EF-1alpha - PIF3 - MTAD - IRES - PhyB - GBD, dual vector of PIF3-MTAD under EF-1 alpha promoter and PhyB-DBD under IRES promoter; See growth conditions below.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||90499||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Growth in Bacteria
Growth Strain(s)NEB Stable
Growth instructionsPick a small colony after streaking or transforming, as plasmid is very unstable. We recommend screening several small colonies by restriction digest before storing immediately upon receipt.
Copy numberHigh Copy
Gene/Insert namemito-tFD and mito-tFNR
- Cloning method Restriction Enzyme
- 5′ cloning site unknown (unknown if destroyed)
- 3′ cloning site unknown (unknown if destroyed)
Additional growth instructions: For larger cultures, the depositing lab minipreps a sample before midiprepping. Perhaps if you are just making glycerol stocks try growing 2 or maybe 3 large cultures and miniprepping a sample to check for the full sized plasmid. They also try to start with a small sized colony and are more stringent about not letting the culture overgrow.
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:pPKm-230 was a gift from Todd Coleman (Addgene plasmid # 90499 ; http://n2t.net/addgene:90499 ; RRID:Addgene_90499)
For your References section:Biosynthesis of Orthogonal Molecules Using Ferredoxin and Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded PhyB Optogenetics. Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP. ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 10.1021/acssynbio.7b00413. Epub 2018 Jan 24. 10.1021/acssynbio.7b00413 PubMed 29301067