Intermediate part consisting of CI promoter followed by tetR, LacI and Tet Operator
This is an intermediate part using the biobrick BBa_K182002 as the upstream part and BBa_K182001 as the downstream part
This part has been created using the BBa_K182001 and BBa_k182002 and contains a CI regulated promoter with TetR, LacI and Ter promoter attached to it. The BBa_k182001 consists of LacI (RBS+, Term+) and a tet promoter and the BBa_K182002 contains a Lambda CI promoter with a tetR protein containning a ribosomal binding site.
We used our earlier submitted two parts available from the registry of standard parts . BBa_K182002 is the upstream part and BBa_K182001 is the downstream part.
Intermediate Part consisting of LacI ( RBS+, Term +) and a Tet operator.
This is an intermediate part using the biobrick BBa_I732820 (RBS, LacI, Term) upstream and the Biobrick BBa_R0040 (TetR repressible promoter) downstream. It comes with a ribosome binding site upstream of LacI and a Terminator downstream (BBa_B0015) There is no Promoter upstream of LacI, meaning that LacI will not be expressed in this piece. The TetR repressible promoter BBa_R0040 is downstream of the Terminator.
We used two parts from the registry of standard parts. BBa_I732820 is the upstream part and BBa_R0040 is the downstream part.
Intermediate Part consisting of a cI regulated promoter and TetR (RBS+, Term-).
This is an intermediate part using the biobrick BBa_R0051 as the upstream part and BBa_S03518 as the downstream part. BBa_R0051 is a cI regulated promoter. The promoter has two binding sites for the cI repressor and is repressed when it binds. BBa_S03518 is a TetR protein with a Ribosome Binding site upstream and an LVA tag attached to its end. TetR binds to TetR regulated promoters such as BBa_R0040. aTc (anhydrotetracycline) binds to TetR causing it to be released from the promoter and reintroducing transcription. This part comes without a terminator.
We used two parts from the registry of standard parts. BBa_R0051 is the upstream part and BBa_S03518 is the downstream part.
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.
A synthetic oscillatory network of transcriptional regulators , Elowitz M.B. , Leibler S., Nature(403),335-38: 2000
The cI regulated promoter is based on the pR promtoer from bacteriohage lambda. The promoter has two two DNA binding sites for lambda cI repressor <bb_part>BBa_C0051</bb_part>. cI binding results in repression of transcription. The specific sequence used here is based on the cI repressible promoter used in the Elowitz repressilator (and references therein).
In order to address concerns about the promoter transcribing in the reverse direction, we have removed the -35 and -10 signals responsible for the promoter activity in the reverse direction. (More details needed here! DE, 2/24/03) Incompatible with host expressing cI repressor.
Vinay S Mahajan, Brian Chow, Peter Carr, Voichita Marinescu and Alexander D. Wissner-Gross
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
tetracycline repressor from transposon Tn10 (+LVA)
Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999)
Coding region for the TetR protein without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission. TetR binds to the pTet regulator (BBa_R0040). aTc (anhydrotetracycline) binds to TetR and inhibits its operation.
References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 BBa_C0040 TetR Protein is based on the TetR sequence from Elowitz's repressilator. It has been modified to include a rapid degradation LVA tail, and includes the BioBrick standard assembly head and tail restriction sites. The RBS has been removed. The stop codon has been changed from TAA to a double stop codon TAATAA.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman.
Conversion of the iGEM parts registry to SBOL2.1