BBa_K784041

BBa_K784041 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K784041
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Inbal Vaknin
Date created: 2012-09-24 11:00:00
Date modified: 2015-05-08 01:13:22

pTetO+ mCherry (FP)



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.

Source

The lab of Roee Amit: http://roee-amit.technion.ac.il/

Sequence Annotation Location Component / Role(s)
pTetO (1)
pTetO (2)
mCherry
RBS
MCS
1,19
26,44
102,812
83,91
816,845
promoter feature/promoter
feature/promoter promoter
CDS feature/cds
ribosome_entry_site feature/rbs
sequence_feature feature/misc
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K784041/1