Team:Groningen/Parts/Submitted Parts

From 2009.igem.org

Submitted Parts

• To get to the partsregistry site of a particular part one can click on the name of the part
• To see what our main findings where (no details or derivations) regarding a particular part one can click on more information

Importers

GlpF
GlpF is an aquaglyceroporin channel that facilitates the transport of As(III). his part has been used to import As(III) into the E.coli cells. It has been used in combination with metal accumulation proteins to make a accumulation device.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190028:Design More Information].

GlpF (with strong RBS)
(GlpF) with the ribosomal binding site .

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190061:Design More Information].

GlpF regulated by medium constitutive promotor
(GlpF with strong RBS) regulated by the constitutive promotor .

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190062:Design More Information].

GlpF regulated by pLacI
(GlpFwith strong RBS) regulated by the inducible promotor

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190063:Design More Information].

HmtA (contains PstI sites)
HmtA (heavy metal transporter A) from Pseudomonas aeruginosa Q9I147 is a P-type ATPase importer. This membrane protein mediates the uptake of copper (Cu) and zinc (Zn). we believe this ATP-driven pump is capable of generating an elevated intracellular concentration of these compounds enabling the harvesting of copper and zinc from the environment.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190018:Design More Information].

Acummulators

fMT
fMT is a metallothionein, binding Arsenite(III) and Arsenate(V), it has higher affinity for As(III). As a metallothionein it is also suitable to bind other metals like zinc, copper or cadmium. This biobrick already contains the ribosomal binding site .

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190019:Design More Information].

fMT regulated by low constitutive promotor
(fMT) regulated by the constitutive promotor .

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190031:Design More Information].

fMT regulated by pLacI
(fMT) regulated by the inducible promotor .

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190032:Design More Information].

MBP-ArsR (fusion protein)
The negative transcriptional regulator ArsR fused with Maltose Binding Protein (MBP) for higher stability. It also carries a His₁₀ tag. It was designed as an accumulator of As(III).

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190027:Design More Information].

Promotors

Arsenic Promoter
Promoter sequence with recognition site for ArsR transcriptional regulator protein. ArsR binds to the promoter sequence in the absence of As and releases on binding of As, thereby activating transcription.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190015:Design More Information].

Arsenic Promoter (with wild type RBS)
Promoter sequence containing the recognition site for ArsR transcriptional regulator protein. Downstream from the promoter is the RBS that is attached to this promoter region in the E. coli genome. ArsR binds to the promoter sequence in the absence of As and releases on binding of As, thereby activating transcription.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190023:Design More Information].

Zinc Promoter
Promoter sequence with recognition site for ZntR transcriptional regulator protein. ZntR activates transcription when Zn(II) is bound.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190016:Design More Information].

Zinc Promoter (with wild type RBS)
The pZntR from E. coli K.12 has a specific RBS site behind it in the genome. Here the RBS site is attachted to the promoter region. The RBS site might influence the activity of the promoter and will be tested in the same way as . ZntR activates transcription when Zn(II) is bound.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190022:Design More Information].

Copper Promoter
Promoter sequence with recognition site for the CueR transcription regulating protein. When copper levels in the cell rise CueR activates transcription.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190017:Design More Information].

Copper Promoter (with wild type RBS)
Promoter sequence with recognition site for the CueR transcription regulating protein and its downstream RBS (as it is found in the E. coli genome). When copper levels in the cell rise CueR activates transcription.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190024:Design More Information].

GVP Constructs

GVP Gas Vesicle Proteins
GVP consists of 6064 base pairs. It's Backbone is With a resistance against Ampicillin/Gentamycin

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_I750016:Design More Information].

GVP regulated by low constitutive promotor
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the constitutive promoter were combined, and the product ligated into vector . The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190025:Design More Information].

GVP regulated by medium constitutive promotor
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the constitutive promoter were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190026:Design More Information].

GVP regulated by pArsR
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the metal sensitive promoter were combined, and the product ligated into vector . The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190033:Design More Information].

GVP regulated by pZntR
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the metal sensitive promoter were combined, and the product ligated into vector . The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190034:Design More Information].

GVP regulated by pCueO
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the metal sensitive promoter were combined, and the product ligated into vector . The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190035:Design More Information].

GVP regulated by pLacI
The Gas Vesicle gene cluster () submitted by Melbourne 2007 team and the LacI and CAP regulated promoter were combined, and the product ligated into vector . The construct was used to test and caracterize the gas vesicle formation.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190036:Design More Information].

Devices

Arsenic Accumulation Device (IPTG induced)
The device consists of a pLacI promoter with fMT (metal binding protein) and GlpF (metal transporter). Upon addition of IPTG the transcription is activated and the fMT gene is expressed in a higher amount compared to the GlpF transporter. Expression of both results in increased metal uptake and accumulation in the cell.

Have a look at the part design page on the registry for [http://partsregistry.org/Part:BBa_K190038:Design More Information].

Arsenic Accumulation Device (constitutive)
The device consists of a constitutive promoter (low expression) with fMT (metal binding protein) and GlpF (metal transporter). Expression of both results in increased metal uptake and accumulation in the cell.