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Team:Imperial College London/Wetlab/BioBricks
From 2009.igem.org
Submitted BioBricks
Summary Table of BioBricks Designed
| Registry Code | Type | Sequence Description |
|---|---|---|
| BBa_K200000 | Coding
| RcsB is a receiver protein which acts as a positive regulator of a number of genes including capsule genes responsible for colanic acid production. |
| BBa_K200001 | Coding
| Dam (DNA Adenine Methylase) The methylase encoded by the dam gene (Dam methylase) transfers a methyl group from S-adenosylmethionine to the N6 position of the adenine residues in the sequence GATC, this protects the DNA from cleavage. |
| BBa_K200002 | Coding
| Colanic acid global regulator ygiV (B3023) increases the production of colanic acid further in conjunction with RcsB by acting as a repressor for mcbR/yncC promoter. YncC/mcbR normally repress colanic acid overproduction so as to increase biofilm formation. |
| BBa_K200003 | Coding
| Waal Ligase is an enzyme responsible for the ligation of an O-antigen to the core oligosaccharide in the Gram-negative bacterium's outer membrane. |
| BBa_K200005 | Coding
| OtsA is the first of two required in the conversion of glucose to trehalose.
This enzyme catalyses the following reaction: UDP-glucose + D-glucose 6-phosphate -> UDP + alpha,alpha-trehalose 6-phosphate |
| BBa_K200006 | Coding
| OtsB This enzyme is the second of two required for the conversion of glucose to trehalose.
This enzyme catalyses the following reaction: alpha,alpha-trehalose 6-phosphate + H2O -> alpha,alpha-trehalose + phosphate |
| BBa_K200007 | Coding
| Cellulase mainly catalyses the reactions that changes crystalline cellulose to cellobiose and then finally to glucose. This cellulase is protease resistant. |
| BBa_K200008 | Coding
| Phenylalanine hydroxylase is the enzyme that breaks down phenylalanine to tyrosine. Deficiency of this enzyme activity results in the autosomal recessive disorder phenylketonuria. |
| BBa_K200013 | Coding
| Opiorphin is a pentapeptide that inhibits the breakdown of endorphines. This results in powerful painrelief. This part contains an enterokinase cleavage site to facilitate synthesis and subsequent activation. |
| BBa_K200014 | Coding
| Opiorphin is a pentapeptide that inhibits the breakdown of endorphines. This results in powerful painrelief. This part contains an enterokinase cleavage site to facilitate synthesis and subsequent activation. This part also contains a HIS tag to facilitate high quality purification. |
| BBa_K200009 | Composite
| Restriction enzyme DpnII is a Type II restriction enzyme that recognises the sequence GATC. Its activity can be blocked by dam methylation. |
| BBa_K200010 | Composite
| Restriction enzyme TaqI is a Type II restriction enzyme that recognises the sequence TCGA. Its activity can be blocked by dam methylation. |
| BBa_K200011 | Composite
| This Lamda cI repressor has a cI857 mutation that results in denaturation of the repressor when the temperature is raised from 30 to 42°C, thereby allowing lambda promoter expression.
When the temperature is raised, typically to 42°C, the functionality of the protein is lost and the cI repressor is no longer able to bind to the operators on its promoter. Therefore, lambda promoter expression increases. |
| BBa_K200012 | Composite
| Lambda promoter (cIts responsive) is different from the common lambda promoter in that it is able to be repressed by the temperature sensitive cI protein (BBa_K200011). When it is not being repressed after 42°C induction, it acts as a strong promoter. |
| BBa_K200017 | Composite
| RBS+OtsB This is an intermediate construct comprising a RBS upstream of the second enzyme for the production of trehalose. A functional transcriptional unit will be obtained upon the ligation of a promoter and the first enzyme (OtsA) upstream of the RBS. To close the unit a double terminator would be required downstream of the OtsB gene. |
| BBa_K200018 | Composite
| pCstA+RBS+GFP+TT This part contains a GFP (green fluorescent protein) reporter in the functional transcriptional unit. As such, activity of the pCstA promoter can be assessed by the expression of GFP. |
| BBa_K200019 | Composite
| pLacI+RBS+RFP+TT+pCstA+RBS+GFP+TT This is our testing construct to assess the two inducible promoters pLacI and pCstA through the expression of reporter molecules RFP and GFP. |
| BBa_K200020 | Composite
| pCstA+RBS This is an intermediate construct that can be ligated to a coding region and terminator to produce a functional transcriptional unit. |
| BBa_K200021 | Composite
| pLacI+RBS This is an intermediate construct that can be ligated to a coding region and terminator to produce a functional transcriptional unit. |
| BBa_K200022 | Composite
| Heat-inducible system with GFP reporter Using the BioBrick part by Harvard '08 (BBa_K098995). We have shown BBa_K098995 to be functional and have improved its characterisation through successful testing with this construct containing a GFP reporter. |
| BBa_K200023 | Composite
| PAH+TT Ligation to a double terminator means that a closed transcriptional unit is produced once a promoter and RBS are ligated before this construct |
| BBa_K200024 | Composite
| RcsB+TT Colanic acid producing gene (RcsB) attached to a double terminator. This is an intermediate construct assembled just prior to promoter ligation. |
| BBa_K200025 | Composite
| pCstA+RBS+RcsB+TT This is the functional transcriptional unit which allows expression of the RcsB receiver protein and positive regulation of colanic acid production. |
| BBa_K200026 | Composite
| RcsB+RBS+GFP+TT This intermediate construct contains a GFP reporter to and can be used to assess the activity of the RcsB receiver protein. Different promoters can be attached upstream of the RBS to complete this transcriptional unit. |
| BBa_K200027 | Composite
| PAH+RBS+GFP+TT (Info here) |
| BBa_K200028 | Coding
| Protease resistant PAH This protease resistant PAH bears a mutation that changes serine 16 into glutamine. This has the effect of mimicking phosphorylation of the protein; modification which has been shown to be associated with protection against proteases (Døskeland et al., 1996) |
| BBa_K200029 | Composite
| pLacI+RBS+RcsB+TT This is the functional transcriptional unit which allows expression of the RcsB receiver protein and positive regulation of colanic acid production. It differs from BBa_K200025, above, by having a different promoter. The differences in functionality between these two constructs will be assessed. |
