Constitutive ER Intein T7 RNAP with YFPH
The promoter is BBa_J23115, the RBS is BBa_B0034, the T7 RNA polymerase is BBa_K145001, the ER Intein is BBa_K1491024, the YFP is BBa_K864100
T7 RNA polymerase that contains the estrogen receptor intein with a constitutive promoter. YFP is added to the end of the plasmid so that the amount of T7 RNA polymerase that is produced can be quantified through fluorescence analysis of YFP.
Constitutive ER Intein T7 RNAP with YFP for quantification of T7 RNAP
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 is BBa_B0034 and SuperYFP2 is BBa_K864100
SuperYFP2 with 6x His tag(BBa_K1491004) and a strong RBS (BBa_B0034). It can be used as a reporter.
T7 RNA polymerase is BBa_K145001 and intein is BBa_K1491024
T7 RNA polymerase with estrogen receptor intein embedded. Non functional with intein. When estrogen is introduced to the system, the intein splices out and links pieces of T7 RNA polymerase together, creating the functional version of the protein.
SuperYFP2 (BBa_K864100) with 6x His tag for protein purification.
Codes for the N-terminus of T7 RNA polymerase (BBa_K145001). This was used to create a T7 polymerase-intein complex.
In order to insert the intein into T7 RNA polymerase, we needed both the N and C terms.
Chong, Shaorong, Kay S. Williams, Chad Wotkowicz, and Ming-Qun Xu. "Modulation of Protein Splicing of the Saccharomyces Cerevisiae Vacuolar Membrane ATPase Intein." Journal of Biological Chemistry 273.17 (1998): 10567-0577.
The intein contains the human E2 estrogen ligand binding domain. When estrogen binds to the receptor, the intein splices out of the protein and joins the N and C terms of the protein, thereby activating it. This allows for an estrogen sensitive protein.
Conversion of the iGEM parts registry to SBOL2.1