The biobricks are all previous submissions. The novel part of this biobrick is the ftsZ gene, biobrick K1011000.
This biobrick enables the complete production of ftsZ and GFP (as an indicator of production). The same RBS is used upstream of both ftsZ and GFP in order to approximate a 1:1 production, so that the flourescent intensity will give a relative indication of ftsZ production.
Double terminator consisting of BBa_B0010 and BBa_B0012
R0011+B0034, strong IPTG-inducible promoter with strong RBS.
This part contains the strong, IPTG-inducible promoter R0011, combined with the strong RBS B0034. This part can be used as a strong protein expression system when combined with a protein coding sequence.
This part should offer greater levels of expression when compared to similar parts using the natural lac promoter (R0010).
The organism for which we based our sequence is Escherichia coli str. K-12 substr. MG1655, located at the following NCBI url: http://www.ncbi.nlm.nih.gov/nuccore/NC_000913.2?report=genbank&from=105305&to=106456
This is an E. coli ftsZ gene. FtsZ is a protein that polymerizes into the Z ring that is formed at the septum site during cellular division. The over expression of ftsZ will lead to non-specific septum sites at the polar ends, which result in the production of E. coli mini-cells.
There is an illegal EcoR1 restriction site at bp 102-107 of the gene. We changed the C at site 107 to a T, which maintained the codon optimization and removed the illegal restriction site.
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Transcriptional terminator consisting of a 64 bp stem-loop.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
Promoter (lacI regulated, lambda pL hybrid)
represillator of Elowitz and Leibler (2000)
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The PLlac 0-1 promoter is a hybrid regulatory region consisting of the promoter P(L) of phage lambda with the cI binding sites replaced with lacO1. The hybrid design allows for strong promotion that can nevertheless be tightly repressed by LacI, the Lac inhibitor (i.e. repressor) (<bb_part>BBa_C0010</bb_part>) ([LUTZ97]). The activity of the promoter can be regulated over a >600-fold range by IPTG in E.Coli DH5-alpha-Z1 (same paper reference).
hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site. LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and increase transcription. This part is incompatible with environments containing lactose or lactose analogs.
Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton
Conversion of the iGEM parts registry to SBOL2.1