Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Xihan Zhang
Date created: 2015-09-16 11:00:00
Date modified: 2015-09-17 09:19:35
pT7-100 + RiboJ + GFP + LVA + 3 terminnators
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K1679040_sequence (Version 1) |
Description
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator. The four T7 RNA polymerases are a family with different activity, the T7 RNA polymerase in this device have approximate relative activity 100%.Notes
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator.We added restriction sites flank these segments, so that users can change this segments into what they prefer. Users can replace T7 with other promoters through SalI and HindIII; replace RiboJ and GFP with other coding sequence through HindIII and StuI.
After transcription, RiboJ becomes a section of functional RNA which comprises the sTRSV- ribozyme which can excise its upstream fragment to reduce noise, and an additional 23-nt hairpin immediately downstream to help expose the RBS. We use it to buffer synthetic circuits from genetic context.
To adapt the protein expression device, three terminators are design to use cooperatively with strong promoters, the LVA degradation tag can accelerate protein degradation rate to decrease cell stress.
Source
Mutated from BBa_k1679026 by PCR.| Sequence Annotation | Location | Component / Role(s) |
| rrnB T2 terminator rrnB T1 terminator T7 terminator LVA GFP rbs RiboJ T7-100 promoter | 18,45 128,214 223,270 293,334 335,1048 1058,1062 1074,1148 1155,1172 | feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/misc tag feature/tag CDS feature/protein ribosome_entry_site feature/rbs ribosome_entry_site feature/rbs feature/promoter promoter |