IPTG Promoter+RBS+EIIA+double terminator
This composite part was designed to upregulate intake of glucose by the cell. Upon further characterization, the preliminary data showed no upregulation of glucose intake. This may be due to the fact that EIIA alone cannot uptake the glucose as it itself is not the membrane protein of the glucose-related sections of the phosphotransferase system (PTS). We hypothesize that upregulating EIIBC (BBa_K1600000), which is a membrane protein complex in addition to EIIA will result in increased glucose uptake.
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).
IIA(Glc) gene from Escherichia coli; double terminator from iGEM Registry
E. coli IIA(Glc) gene (BBa_K1508002) followed by double terminator (BBa_B0015)
Construct was assembled by suffix insertion, namely BBa_B0015 fragment was inserted into BBa_K1508002 backbone.
Fahmi Dwilaksono, Indah Nurulita, Rian Adha Ardinata, Mochammad Isro Alfajri, Arief Budi Witarto
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.
Double terminator consisting of BBa_B0010 and BBa_B0012
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
IIA(Glc) - E.coli glucose-specific phosphotransferase enzyme IIA
Phosphotransferase system (PTS) is a major carbohydrate active -transport system, which is executed by a cascade of protein rally. For glucose uptake, IIA(Glc) is the only enzyme specific to glucose transport. We measure the concentration of glucose in the medium using catabolite repression mechanism toward lacI promoter activity upstream of a reporter gene, mRFP. We expect by over-expressing IIA(Glc) in E. coli, we will obtain different range of glucose standard curve in our measurement system.
The gene coding for IIA(Glc) from E.coli - crr - was isolated by colony PCR using forward primer which contains with restriction sites of EcoRI and XbaI and reverse primers which contains restriction site for SpeI and a double stop codon.
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.
Conversion of the iGEM parts registry to SBOL2.1