Team:Groningen/Parts/Submitted Parts

Submitted Parts

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Importers
 BBa_K190028 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.

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 BBa_K190061 GlpF (with strong RBS) BBa_K190028 (GlpF) with the ribosomal binding site B0034.

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 BBa_K190062 GlpF regulated by medium constitutive promotor BBa_K190061 (GlpF with strong RBS) regulated by the constitutive promotor BBa_J23106.

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 BBa_K190063 GlpF regulated by pLacI BBa_K190061 (GlpFwith strong RBS) regulated by the inducible promotor BBa_R0010

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 BBa_K190018 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.

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Acummulators
 BBa_K190019 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 B0034.

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 BBa_K190031 fMT regulated by low constitutive promotor BBa_K190019 (fMT) regulated by the constitutive promotor BBa_J23109.

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 BBa_K190032 fMT regulated by pLacI BBa_K190019 (fMT) regulated by the inducible promotor BBa_R0010.

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 BBa_K190027 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).

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Promotors
 BBa_K190015 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.

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 BBa_K190023 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.

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 BBa_K190016 Zinc Promoter Promoter sequence with recognition site for ZntR transcriptional regulator protein. ZntR activates transcription when Zn(II) is bound.

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 BBa_K190022 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 BBa_K190016. ZntR activates transcription when Zn(II) is bound.

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 BBa_K190017 Copper Promoter Promoter sequence with recognition site for the CueR transcription regulating protein. When copper levels in the cell rise CueR activates transcription.

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 BBa_K190024 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.

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GVP Constructs
 BBa_I750016 GVP Gas Vesicle Proteins GVP consists of 6064 base pairs. It's Backbone is BBa_J61035 With a resistance against Ampicillin/Gentamycin

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 BBa_K190025 GVP regulated by low constitutive promotor The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the constitutive promoter BBa_J23109 were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.

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 BBa_K190026 GVP regulated by medium constitutive promotor The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the constitutive promoter BBa_J23106 were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.

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 BBa_K190033 GVP regulated by pArsR The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the metal sensitive promoter BBa_K190015 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 More Information.

 BBa_K190034 GVP regulated by pZntR The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the metal sensitive promoter BBa_K190016 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 More Information.

 BBa_K190035 GVP regulated by pCueO The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the metal sensitive promoter BBa_K190017 were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.

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 BBa_K190036 GVP regulated by pLacI The Gas Vesicle gene cluster ( BBa_I750016 ) submitted by Melbourne 2007 team and the LacI and CAP regulated promoter BBa_R0010 were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.

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Devices
 BBa_K190038 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.

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 BBa_K190039 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.

Have a look at the part design page on the registry for More Information.