BBa_K1679039

BBa_K1679039 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1679039
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 09:03:49

pT7-10 + RiboJ + GFP + LVA + 3 terminnators



Types
DnaRegion

Roles
engineered_region

Device

Sequences BBa_K1679039_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 10%.

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

Synthesized by Genewiz.

Sequence Annotation Location Component / Role(s)
rrnB T2 terminator
rrnB T1 terminator
T7 terminator
LVA
GFP
rbs
RiboJ
T7-10 promoter
18,45
128,214
223,270
293,334
335,1048
1058,1062
1074,1148
1155,1172
feature/misc sequence_feature
feature/misc sequence_feature
sequence_feature feature/misc
feature/tag tag
CDS feature/protein
ribosome_entry_site feature/rbs
ribosome_entry_site feature/rbs
promoter feature/promoter
igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1679039/1