Types | DnaRegion
|
Roles | Reporter
engineered_region
|
Sequences | BBa_K784041_sequence (Version 1)
|
Description
This part includes 2 inducible promoters of pTetO (activated by tetracycline), for reducing the leakiness of this promoter, and a red fluorescence protein (mCherry). This part was donated to us by Roee Amit, and was sequence confirmed by him. The part is present in plasmid pSB1C3, with the standard prefix and suffix, but the sequence of the part contains EcoRI and PstI restriction sites, so the optimal solution will be using this part in Gibson assembly reaction.
This part places mCherry downstream to the pTetO promoter. Reported activities of the promoters are given in strain Top10 grown in LB media. The activity of this promoter in this bacteria strain is constitutive and the fluorescence measure results can be seen at the "results for this part" section.
This part can be used for inducible/constitutive expression (depends on the bacteria's strain) of the FP, and can be combined with other genes.
Cloning this part was done by amplifying the promoter and the FP together, adding XbaI site in the 5' and SpeI in the 3' ends, for cloning it to the registry plasmids.
Notes
This part have EcoRI and PstI restriction sites, this will be a problem when trying to clone this part by restriction to the registry plasmids. An alternative solution can be the Gibson assembly reaction, which doesn???t require digestion at all, no matter what is the restriction site is, you can clone it to any of the registry plasmids.