Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Thomas Mathijs Swartjes
Date created: 2016-09-29 11:00:00
Date modified: 2016-10-14 08:38:26
434- and lambda cI balance operon + mRFP reporter
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K1913006_sequence (Version 1) |
Description
This part contains two viral genes: 434- and lambda cI (BBa_C0052 and part of BBa_K081007) expressed under the pBAD/AraC promoter (BBa_I0500). The 434 cI gene is not yet preceded by an RBS, allowing tuning of the cI protein balance. The 434 cI protein encoded in this part contains a C-terminal LVA tag causing more rapid protein degradation, the lambda cI gene in this part does not encode an LVA tag.Additionally this part contains an operon where mRFP (BBa_K081014) is expressed under a modified lambda cI promoter (BBa_I12006). This promoter is (according to the parts page) induced by lambda cI and repressed by 434 cI. This last operon also includes a LVA-tagged lambda cI (BBa_K081007) encoding gene, in theory establishing positive feedback of the modified lambda cI promoter.
The two operons in this part were also submitted as separate parts:
- 434- and lambda cI operon: BBa_K1913007
- 434- and lambda cI balance RFP reporter: BBa_K1913016
The complete part is intended to respond with mRFP expression upon a change in strength of the inducible promoter (BBa_I0500). When - after a period of induction with L-Arabinose - the promoter is repressed with glucose, one expects an increase in mRFP (BBa_K081014) expression.
Notes
I chose to digest the mRFP operon with SpeI and PstI and ligate it into pSB1C3 with the Arabinose inducbile operon digested with XbaI and PstI. This would (and did) result in a plasmid that contains both operons in the most understandable order and colours successful clones light-red.Source
The part has been assembled from parts BBa_K1913007 and BBa_K1913016 via digestion a subsequent ligation. BBa_K1913007 and BBa_K1913016 were both assembled through Gibson assembly from fragments derived from part in the iGEM registry.| Access | Instance | Definition |
| public public | BBa_K1913007 BBa_K1913016 | BBa_K1913007 BBa_K1913016 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1913007 BBa_K1913016 | 1,2771 2780,4447 | BBa_K1913007 BBa_K1913016 |