Team:ITESM Mexico/Assembly




Igem Itesm


Mechanism con NOMBRE RecA.jpg

Part1.jpg Assembly for the photoreceptor
Part2.jpg Assembly for high and low concentration

caption BBa_J23109: Constitutive promoter family. Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library. J23119 is the "consensus" promoter sequence and the strongest member of the family. All parts except J23119 are present in plasmid J61002. Part J23119 is present in pSB1A2. This places the RFP downstream of the promoter. Reported activities of the promoters are given as the relative fluorescence of these plasmids in strain TG1 grown in LB media to saturation. See part J61002 for details on their use. These promoter parts can be used to tune the expression level of constitutively expressed parts. The NheI and AvrII restriction sites present within these promoter parts make them a scaffold for further modification. Ampicillin resistant.
caption BBa_B0034: RBS based on Elowitz repressilator. Ampicillin resistant.
caption BBa_I15008: One of two requisite genes required for the biosynthesis of phycocyanobilin from heme. Usage and Biology Ho1 oxidizes the heme group using a ferredoxin cofactor, generating biliverdin IXalpha and representing the first of two steps in phycocyanobilin (PCB) biosynthesis. PCB associates with Cph8, creating a light responsive protein complex. Functions in tandem with BBa_I15009 for PCB biosynthesis. PCB then associates withPart:BBa_I15010, a light responsive Cph8/EnvZ fusion protein. Kanamycin resistant.
caption BBa_I15009: Phycocyanobilin:ferredoxin oxidoreductase (PcyA) from synechocystis Second of two required phycocyanobilin biosynthetic genes. Usage and Biology: Converts biliverdin IXalpha (BV)(produced by Part:BBa_I15008) to phycocyanobilin (PCB), the immediate precursor of cyanobacterial phytochromes. Associates with Part:BBa_I15010 to transduce a light signal into a genetic response. Kanamycin resistant.
caption BBa_K225000: Green light receptor. Fusion protein of light responsive domains of CcaS from Synechocystis sp. PCC 6803 and histidine kinase EnvZ. For the use as a green light activated actuator in E. coli. Usage and Biology: GlrN is the fusion protein of light responsive domains of CcaS and EnvZ designed by changing the histidine kinase region of CcaS to EnvZ domain of cph8 (BBa_I15010).

  • Requires phycocyanobilin biosynthetic genes (BBa_I15008 and BBa_I15009) for chromophore formation.
  • Must be used in E. coli deficient in natural EnvZ.

Expecting function (without experimental proof): Exposure to green light (535 nm) phosphorylate EnvZ histidin kinase domain. The activated EnvZ phosphorylates endogenous OmpR, a transcription factor which activates transcription from the OmpR positive promoter (BBa_R0083). Ampicillin resistant.

caption BBa_B0015: Double terminator consisting of BBa_B0010 and BBa_B0012. Ampicillin and Kanamycin resistant.
caption BBa_R0083: Promoter (OmpR, positive) Positively regulated, OmpR-controlled promoter. This promoter is derived from the upstream region of ompC. Phosphorylated OmpR binds to the operator site and activates transcription. This is a truncated version of BBa_R0082. Usage and Biology: This promoter contains upstream regions of the ompC porin gene. The regulation of ompC is determined by the EnvZ-OmpR osmosensing machinery. EnvZ phosphorylates OmpR to OmpR-P. At high osmolarity, EnvZ is more active, creating more OmpR-P. OmpR-P then binds to the low-affinity OmpR operator sites upstream of ompC. Activation of ompC has been experimentally shown to occur with just the C1 OmpR binding region, provided that the C1 region is placed next to the -35 signal (Maeda and Mizuno, 1990). This promoter is a truncated version of BBa_R0082, which contains C1, C2, and C3. Ampicillin resistant.
caption BBa_K587001: RecA protein coding sequence Ampicillin resistant.
caption BBa_C0051: cI repressor from E. coli phage lambda (+LVA) Coding region for the cI repressor based on cI repressor from bacteriophage lambda modified with an LVA tail for rapid degradation of the protein. cI repressor binds to the cI regulator (BBa_R0051). Ampicillin resistant.
caption BBa_K079043: Lambda OR3 - operator library member Lambda Genetic switch is a complex and deeply characterized system. Ampicillin resistant.
caption BBa_K079042: Lambda OR2 - operator library member. Lambda Genetic switch is a complex and deeply characterized system. Ampicillin resistant.
caption BBa_K079041: Lambda OR1 - operator library member. Lambda Genetic switch is a complex and deeply characterized system. Ampicillin resistant.
caption BBa_K206000: pBAD strong: Inducible promoter that responds to low arabinose concentrations. Click on the part's number for description.
caption BBa_K587002: Crxst- Lock sequence for concentration mechanism Ampicillin resistant.
caption BBa_K145015: GFP with LVA tag Green Fluorescent Protein with LVA tag for rapid degradation. This part was thoroughly tested by the 2008 KULeuven team. This team showed that this GFP mutant is degraded at least 35x faster than GFP (mut3b) and that it has a half life of about 74 minutes. Ampicillin resistant.
caption BBa_K587003: Tast - Key to unlock crxst. Ampicillin resistant.
caption BBa_K206001: Name: pBAD weak Input: L-arabinose

Output: PoPS Usage and Biology: pBAD is an E.coli promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC (BBa_I13458) binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription. K206001 is a variant pBAD promoter with a modified AraI1 site that has been shown to be less responsive at low concentrations of arabinose and to exhibit a lower maximum expression than the wild type (BBa_I13453), as measured by coupling to a fluorescent reporter. What you can do with it: At a given level of arabinose input, BBa_K206001 will provide a lower level of PoPS output than its family members, allowing analog device responses. See our wikifor a project that makes use of this property. Compatibility: Chassis: Best used in the E. coli strain BW27783, which has been modified to permit homogeneous pBAD promoter expression by substituting the chromosomal arabinose-dependent promoter of AraE (arabinose transporter protein) with a constitutive promoter . Backbone: Has been shown to work on plasmid pSB1C3. Reporter: Has been shown to work with reporters BBa_I13507 and BBa_I763020. Ampicillin resistant.

caption BBa_K587006: Crxwk. Ampicillin resistant.
caption BBa_E0022: Enhanced cyan fluorescent protein derived from A. victoria GFP Cyan fluorescent protein (ECFP) reporter coding sequence without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission. Ampicillin resistant.
caption BBa_K587004: Tawk. Key to unlock crxwk. Ampicillin resistant.
caption BBa_K587005: iTast - inhibits the action of tast. Ampicillin resistant.