Promoter Tester 6803 Native RBS sigA Reference Dual Luciferase Assay
Synechocystis sigA promoter controlled Renilla Luciferase
psbA2 5'UTR Native RBS + Firefly Luciferase + Terminator
5'UTR and native RBS from psbA2 in Synechocystis sp. PCC 6803
Genomic sequences from Synechocystis sp. PCC 6803
This is the 5'UTR and RBS region from gene psbA2 in Synechocystis sp. PCC 6803. It covers the entire region from the end of the promoter (as reported in Imamura S and Asayama M. 2009. Sigma Factors for Cyanobacterial Transcription. Gene Regulation and Systems Biology 3: 65-87) to immediately before the start codon of the psbA2 gene. This part can serve as a general RBS for Synechocystis sp. PCC 6803, until a library of RBS strengths in PCC 6803 is assembled. The psbA2 gene codes for the photosystem II D1 protein in this species, and therefore we assume (and plan to verify) that the RBS strength is moderate to strong, due to the high requirement for this protein in photosynthesis.
We decided to go with the full region from the promoter to the start codon. This means that, with the addition of the biobrick scar, the RBS may be located farther from the start codon then is optimal. We plan to determine optimum spacing of the RBS in Synechocystis sp. PCC 6803 in future work.
Double terminator consisting of BBa_B0010 and BBa_B0012
Firefly luciferase - luciferase from Photinus pyralis
This firefly luciferase comes from Promega PGL3 vector.
Firefly luciferase is luciferase from Photinus pyralis. By oxidation of substrate luciferin light is produced. Luciferase thus acts as a reporter protein when connected to other proteins or promoters.
Part is cloned into pSBAK3 vector with terminator.
luciferase: luciferin 2-monooxygenase from Renilla reniformis (EC 1.13.12.5; SwissProt:: P27652)
Rluc is a protein called Renilla`s luciferase and it emits light when adding the right substrate. That is why it can bi used as a reported to track other proteins or to monitor the activity of a promotor.
Rluc is cloned in BioBrick vector pSB1AK3.
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