RFP and eCDM8 expression + Theophylline riboswitch tetracycline resistance protein
RFP expression followed by high strength promoter (J119031) and a high strength C dog RBS (J119028) controlling expression of the eCDM8 gene (J100100). The clone also contains theophylline riboswitch D (Part J100065) and tetracycline resistance protein (Part J119140).
Contains an IPTG inducible promoter an RBS, RFP, and terminator.
eCDM8 expression + Theophylline riboswitch tetracycline resistance protein
A high strenght promoter (J119031) and a high strength C dog RBS (J119028) control expressin of the eCDM8 gene (J100100). The clone also contains theophylline riboswitch D (Part J100065) and tetracycline resistance protein (Part J119140).
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.
**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)
Campbell et al., PNAS v99 p7877 URL
monomeric RFP: Red Fluorescent Protein. Excitation peak: 584 nm Emission peak: 607 nm
TAATAA double stop codon added (DE). Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
Theophylline Riboswitch - tetracycline resistance protein
Constructed using BsaI GGA of PCR products from J100065 and J31007.
This device contains a synthetic riboswitch that can be used in E. coli. In the presence of theophylline, the transcription of the TetA gene occurs, but in the absence of theophylline, very little transcription takes place. This device contains a modified T5 promoter, which, in the absence of cymR, acts as a strong constitutive promoter. The ribosomal binding site is contained in the riboswitch. This riboswitch is modified from riboswitch D as described in the paper by Topp et al. References: Shana Topp, Colleen M. K. Reynoso, Jessica C. Seeliger, Ian S. Goldlust, Shawn K. Desai, Doroth??e Murat, Aimee Shen, Aaron W. Puri, Arash Komeili, Carolyn R. Bertozzi, June R. Scott, and Justin P. Gallivan. Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species. Applied and Environmental Microbiology, 76:23, 7881-7884. December 2010.
BsaI sites were introduced into PCR products, but are not present in the final construct.
eCDM8 expression with high promoter and high C dog RBS
A low promoter, J119030 with a low c.dog J119025, this combination is known as S05011. The eCDM8 gene (J100100) was then inserted into the S05011 vector creating a pSB1A8 plasmid with a low promoter, low c.dog, and the eCDM8 gene.
Conversion of the iGEM parts registry to SBOL2.1