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Team:UNITN-Trento/Parts
From 2012.igem.org
Our Parts
Here you can find our favourite Parts and all the Parts we built. Be also sure to check out our Team Parts Page on the Registry website!
FAVORITES
K731030 - Inducible araC-pBAD promoter regulating M256I CysE
The CysE M256I gene (K731010) is here regulated by the araC-pBAD promoter (K731201), which is inducible by arabinose. This part was sub cloned and characterized in psB3C5.
Registry PageK731400 - IPTG inducible Cysteine desulfhydrase (CysDes)
This part is composed of K731300 and K731600. It is an IPTG inducible device regulating the expression of a Cysteine Desulfhydrase from Treponema Denticola (CysDes). This enzyme is an aminotransferase that converts cysteine into pyruvate, ammonia and hydrogen sulfide. The part has been successfully operated and characterized in psB1C3 and psB4K5.
Registry PageK731710 - Platform for terminators analysis with E. coli RNA polymerase by fluorimetric assay
This part is used to analyse terminators: two subsequent fluorescent proteins (mCherry and A206K Venus), separated by Prefix-Suffix linker, allow standardization of terminator's presence effect. The vector was obained by mutagenesis of BBa_K731700, replacing T7 promoter with pTAC. It contains Ampicillin resistance gene and is inducible by IPTG (pTAC-lacO). The combined use of BBa_K731710 and BBa_K731700 allow also to analyse any potential difference in terminators' activity due to different polymerases.
Registry PageAll our other Parts
K731000 - CysE (Serine Acetyltransferase)
CysE is an enzyme involved in cysteine biosynthesis, also known as SAT. It catalyzes the acetylation of serine to give O-acetylserine, a precursor for the biosynthesis of cysteine. Some O-acetylserine is also converted to N-acetylserine, which in turn triggers the assimilation of sulfate through specific genes.
K731010 - M256I CysE (Serine Acetyltransferase)
CysE is an enzyme involved In cysteine biosynthesis, also known as SAT. It catalyzes the acetylation of serine to give O-acetylserine, the CysE final precursor. Some O-acetylserine is also converted to N-acetylserine, which in turn triggers the assimilation of sulfate through specific genes. M256I CysE shows enhanced secretion of cysteine and is not inhibited by Cysteine overproduction.
K731020 - araC-pBAD regulated WT CysE
The CysE gene (K731000) is here regulated by the araC-pBAD promoter (K731201), which is inducible by arabinose. This part was sub cloned and characterized in psB3C5.
K731040 - araC-pBAD regulated sfGFP-tagged M256I CysE
A sfGFP tagged M256I CysE inducible by arabinose. This part was subcloned and characterized in pSB3C5.
K731201 - Inducible araC-pBAD promoter
This E. coli promoter is positively regulated by arabinose. araC is the regulator protein: in absence of arabinose the protein binds to araI1 and araO2 and the DNA forms a loop that prevents RNA polymerase from binding the promoter. When arabinose is added to the culture it binds to araC, disabling the interaction with araI1 and araO2 sites, and also allowing it to bind araI site, promoting transcription. Promoter activity is also influenced by glucose levels: in its absence cAMP levels are high and the the complex cAMP+CAP (catabolite activator protein) binds to lacI promoting transcription. In presence of glucose, cAMP levels are low and promoter activity is inhibited.
K731250 - Inducible araC-pBAD promoter with GFPmut3b, strong RBS
This part has been used for testing and characterization of araC-pBAD promoter (BBa_K731201). The part is composed of part BBa_K731201 and BBa_E0840. This part was successfully operated and characterized both in psB1C3 and psB3C5.
K731255 - Inducible araC-pBAD promoter with GFPmut3b, weak RBS
This part has been used for testing and characterization of araC-pBAD promoter (part BBa_K731200) when using a weak RBS. The part is composed of part BBa_K731200 and BBa_E0240.
K731300 - LacI + LacIq promoter (reverse)
This transcriptional regulator includes the LacI repressor and its constitutive promoter LacIq which operates in the reverse direction. The LacI repressor protein is inhibited by IPTG, and it is a very useful device to control and regulate the expression of a gene of interest.
K731400 - IPTG inducible Cysteine desulfhydrase (CysDes)
This part is composed of K731300 and K731600. It is an IPTG inducible device regulating the expression of a Cysteine Desulfhydrase from Treponema Denticola (CysDes). This enzyme is an aminotransferase that converts cysteine into pyruvate, ammonia and hydrogen sulfide. The part has been successfully operated and characterized in psB1C3 and psB4K5.
K731500 - Inducibile tac promoter and lac operon
This is E.coli lactose-inducibile promoter. lac repressor (encoded from lacI gene) is the regulator protein: in absence of lactose the protein binds to lac operator, preventing RNA-polymerase binding and transcription. Low, constitutive levels of transcription can be seen anyhow.
K731520 - Lactose-inducible sfGFP
This is a construct carrying the sfGFP gene with low-efficiency RBS, under lac operator control. lac repressor (encoded from lacI gene) is the regulator protein: in absence of lactose the protein binds to lac operator, preventing RNA-polymerase binding and transcription.Low, constitutive levels of transcription can be seen anyhow.
K731600 - Cysteine Desulfhydrase (CysDes)
This part encode for a cysteine desulfhydrase from Treponema denticola, which is known to be a strong enzyme that produces H2S, ammonia and pyruvate starting from cysteine. It is often indicated also as Hly.
K731700 - T7 Platform
This part is used to analyse terminators: two subsequent fluorescent proteins (mCherry and A206K Venus), separated by Prefix-Suffix linker, allow standardization of terminator's presence effect. The vector is a modified pET21b; it contains Ampicillin resistance gene and is inducible by IPTG (T7promoter-lacO). The combined use of BBa_K731710 and BBa_K731700 allow also to analyse any potential difference in terminators' activity due to different polymerases.
K731701 - BBa_K731700 with T7 terminator
T7 terminator T7 polymerase testing construct.
K731702 - BBa_K731700 with BBa_K731722 terminator
E. Coli terminator T7 polymerase testing construct.
K731711 - BBa_K731710 with T7 terminator
T7 terminator coli polymerase testing construct.
K731712 - BBa_K731710 with BBa_K731722 terminator
E. coli terminator coli polymerase testing construct 2.
K731712 - BBa_K731710 with BBa_K731722 terminator
E. coli terminator coli polymerase testing construct 2.
K731721 - T7 terminator
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K731722 - B0010 Coli terminator
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