mCherry & GFP expression device for testing a biological virtual machine
mCherry expression is controlled by an e. coli promoter and ribosome binding site. GFP expression is controlled by an orthogonal promoter and ribosome binding site.
GFP is downstream of mCherry because the e. coli terminator is very efficieint (>95%) and the T7 terminator downstream of GFP is only ~80% efficient.
Reporter device for dedicated transcription and translation
T7 promoter with an A->C mutation at the -16 position of the consensus sequence
This is a theoretical intermediate part that will never be physically built, but is necessary for creating the part files of my RBS measurement devices which do not use Biobricks Cloning Sites for inserting the RBS sequence. This is the region found downstream of the RBS sequence in all of the mCherry measurement devices.
This is a theoretical intermediate part that will never be physically built, but is necessary for creating the part files of my RBS measurement devices which do not use Biobricks Cloning Sites for inserting the RBS sequence. This is the region found upstream of the RBS sequence in all of the GFP and mCherry measurement devices, including the AvrII/XbaI scar site that is created between this part and the RBS during cloning.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
T7 gene 1 specific transcriptional terminator
5 prime UTR from the OmpA gene in E.coli used for stabilizing the downstream RNA transcript
This region was PCR amplified from the genome of MG1655.
3' stabilizing mRNA designed by Christina Smolke and Jay Keasling for stabilizing gfp.
A single base pair change (A to T at position 25) from the original design was made in order to remove a PstI site occurring in the loop of the hairpin. This should have no effect on the secondary structure.
This is the 6 bp mixed site that results when ligating the compatible ends (CTAG 5' overhang) that result from AvrII cleavage (CCTAGA) of the upstream part and XbaI (TCTAGA) cleavage of the downstream part.
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.
mCherry from Roger Y. Tsien's lab, altered to be BioBrick compatible
mRFP (DsRed) derived, altered to be a biobrick by removing a PstI site and adding in the ends.
Made a point mutation to eliminate a PstI site in the middle and then added BioBrick ends using PCR. Sequenced using primers that bind to the pSB1A2 plasmid on either side of the brick.
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