Team:Tacoma RAINmakers/Parts

Team:ECUST/Lab/Notebook

Our Biobricks

Parts Overview

The Tacoma RAINMakers have built two biological arsenic circuits that use a combination of native E. coli and coral genes to detect arsenic ions in solution. The circuits vary in which reporter they produce, either the chromoprotein spisPink (from S. pistillata) or amilCP (from A.millepora). Each circuit contains an Arsenic regulator known as ArsR. The ArsR protein binds to a promoter with an ArsR binding site (PArsR) and inhibits the transcription of the downstream reporter gene. Only in the presence of a transcription factor, which in our case is arsenic, does the reporter become derepressed and the reporter is expressed. This circuit allows us to control chromoprotein expression using a protein sensitive to, and capable of binding with, arsenic ions.

Submitted Parts

BBa_K2673000 pSB1C3/PcArsR_PArsR-spisPink

Part Type: Composite
Description: pSB1C3/PcArsR_PArsR-spisPink is an arsenic detection circuit. It is a two-part circuit made up of a series of promoter-gene sequences. The first part of the circuit, PcArsR, contains an E. coli-derived constitutive promoter (Pc), a Ribosome Binding Site, Arsenic Regulator (ArsR), and a Double Terminator. This sequence creates unregulated expression of ArsR. The second part of the circuit contains an E. coli-derived Arsenic Regulator promoter (PArsR), a Ribosome Binding Site, the spisPink chromoprotein, and a Double Terminator. The PArsR sequence has a binding site for ArsR, which blocks expression of the reporter in the absence of Arsenic ions. When Arsenic ions are present the spisPink chromoproteins are produced.
Website: http://parts.igem.org/Part:BBa_K2673000

BBa_K2673001 pSB1C3/PcArsR_PArsR-amilCP

Part Type: Composite
Creator: Tacoma RAINmakers
Description: pSB1C3/PcArsR_PArsR-amilCP is an arsenic detection circuit. It is a two-part circuit that combines two series of promoter-gene sequences on one plasmid. The first part of the circuit, PcArsR, contains an E. coli-derived constitutive promoter (Pc), a Ribosome Binding Site, Arsenic Regulator (ArsR), and a Double Terminator. This sequence allows for unregulated expression of ArsR. The second part of the circuit contains an E. coli-derived Arsenic Regulator promoter (PArsR), a Ribosome Binding Site, the amilCP chromoprotein, and a Double Terminator. The PArsR sequence has a binding site for ArsR which blocks expression of the reporter in the absence of Arsenic ions. When Arsenic ions are present amilCP chromoproteins are produced.
Website: http://parts.igem.org/Part:BBa_K2673001

BBa_K2673002 pSB1C3/PArsR-spisPink

Part Type: Composite
Creator: Tacoma RAINmakers
Description: pSB1C3/PArsR-spisPink is used to test part of the complete two-part arsenic sensing circuit. This part contains an E. coli-derived Arsenic Regulator promoter (PArsR), a Ribosome Binding Site, the spisPink chromoprotein, and a Double Terminator. The expression of the spisPink chromoprotein is regulated only if ArsR is present. In this plasmid the chromoprotein expression is unregulated.
Website: http://parts.igem.org/Part:BBa_K2673002

BBa_K2673003 pSB1C3/PcArsR

Part Type: Composite
Creator: Tacoma RAINmakers
Description: The part contains a Ribosome Binding Site, PcArsR, and a Double Terminator. This sequence creates unregulated expression of ArsR. There is no reporter.
Website: http://parts.igem.org/Part:BBa_K2673003

BBa_K2673004 pSB1C3/PArsR-amilCP

Part Type: Composite
Creator: Tacoma RAINmakers
Description: pSB1C3/PArsR-amilCP is used to test part of the complete two-part arsenic circuit. This part contains an E. coli-derived ArsR promoter (PArsR), a Ribosome Binding Site, the spisPink chromoprotein, and a Double Terminator. The expression of the amilCP chromoprotein is regulated only if ArsR is present. In this plasmid the chromoprotein expression is unregulated.
Website: http://parts.igem.org/Part:BBa_K2673004

BBa_K2673005 pSB1C3/T7-amilCP

Part Type: Composite
Creator: Tacoma RAINmakers
Description: To test the AmilCP promoter, we created a seperate circuit consisting of the AmilCP reporter an a high expression T7 promoter. This should produce unregulated chromoprotien, however in lab testing proved unsuccessful. We have speculated that this is due to a lack of sequence spacing around the promoter, but have yet to test this theory.
Website: http://parts.igem.org/Part:BBa_K2673005

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