KaiA and KaiBC is regulated by different promoter in order to generate different amount of Kai proteins at a ratio similar to that measured in vivo KaiA:KaiB:KaiC=1:1:4 (by weight) 1:2.8:2.2 (by amount)
1. M. Nakajima et al., Science 308, 414 (Apr 15, 2005).
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.
The Plac insert was PCR'd from the MG1655 strain of E.coli K12.
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG.
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 this regulator. This part is incompatible with environments containing lactose or lactose analogs.
Transcriptional terminator consisting of a 64 bp stem-loop.
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
KaiA coding region with an extra stop codon at the end and two silent mutations at the EcoRI and PstI cut sites. From Prochlorococcus (Synechococcus) elongatus PCC7942 genome. The actual DNA is synthetic in nature; was synthesized from GeneArt AG. Note that the coding region is bounded by the BioBricks Primers and is on a non-BioBrick vector pGA4 (ampR). The biobricks primers can be cut out along with the KaiA coding sequence with HindIII and SacI. Length of entire plasmid is 3802bp.
KaiB coding region with an extra stop codon at the end. From Prochlorococcus (Synechococcus) elongatus PCC7942 genome. The actual DNA is synthetic in nature; was synthesized from GeneArt AG. Note that the coding region is bounded by the BioBricks Primers and is on a non-BioBrick vector pPCR-Script (ampR). The biobricks primers can be cut out along with the KaiB coding sequence with KpnI and SacI. The length of the entire plasmid is 3220bp.
Zhipeng Sun, Hetmann Hsieh, Jeffrey Lau, David Ramos
KaiC coding region with an extra stop codon at the end. Two silent point mutations at EcoRI and PstI site. From Prochlorococcus (Synechococcus) elongatus PCC7942 genome. The actual DNA is synthetic in nature; was synthesized from GeneArt AG. Note that the coding region is bounded by the BioBricks Primers and is on a non-BioBrick vector pGA4(ampR). The biobricks primers can be cut out along with the KaiCcoding sequence with KpnI and XhoI. The length of the entire plasmid is 4558bp.
Zhipeng Sun, Hetmann Hsieh, Jeffery Lau, Dave Ramos
Conversion of the iGEM parts registry to SBOL2.1