Part designed for verification and characterization of the FUR/LacI inverter
E. coli constitutive promoter with Ferric Uptake Regulator & LacI Inverter. Activates RFP expression in presence of iron
LacI and FUR proteins are optimized for Acidithiobacillus ferrooxidans. It has two RBS after the constitutive promoter and before the FUR CDS.
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.
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
The RBS used is the BBa_B0034, the terminator is the BBA_B0012, the LacI reculated Promoter is the BBa_R0011. The sequence for the FUR Box was obtained from the results of the paper of Quatrini et al. (Citation in our wiki), choosing the Fur box with the best score. The sequence for the FUR protein was obtained from the Acidithiobacillus ferrooxidans genome.
Inverter with Ferric Uptake Regulator & LacI. Activates gene expression in presence of iron. LacI and FUR proteins are optimized for Acidithiobacillus ferrooxidans.
Carmen Padilla, Melissa Gonzalez & Melissa Rios
The sequence for the FUR Box was obtained from the results of the paper of Quatrini et al. (Citation in our wiki).
Ferric Uptace Regulator Protein codon-optimized for Acidithiobacillus Ferrooxidans, the FUR Protein in presence of iron will block the transcription because it will bind to its FUR Box.
You will have to add a FUR Box to repress the transcription with this FUR protein in presence of iron.
It is the FUR Box with the highest score in the work of: Quatrini, R., Lefimil, C., Veloso, F. A., Pedroso, I., Holmes, D. S., & Jedlicki, E. (2007). Bioinformatic prediction and experimental verification of Fur-regulated genes in the extreme acidophile Acidithiobacillus ferrooxidans. Nucleic Acids Research, 35(7), 2153???2166. http://doi.org/10.1093/nar/gkm068
This is a FUR Box with high affinity to BBa_K2055009. In presence of iron, it will block the downstream transcription.
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.
**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)
Campbell et al., PNAS v99 p7877 URL
monomeric RFP: Red Fluorescent Protein. Excitation peak: 584 nm Emission peak: 607 nm
TAATAA double stop codon added (DE). Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
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