Team:Lambert GA/Parts

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PARTS DESCRIPTION

PHO PARTS DESCRIPTION

BBa_K116401

External Phosphate Sensing Promoter Sequence

The External Phosphate Sensing Promoter is a basic, regulatory part created by iGEM08_NYMU-Taipei. The inducible promoter is activated by the binding of phosphorylated PhoB transcription factor from the Pho Regulon signaling pathway.

BBa_K2447000

Extracellular phosphate sensor with GFP reporter (PhoB)

BBa_K2447000, a composite part created by NUS Singapore iGEM 2017 as an improvement to BBa_K116404, is an extracellular phosphate sensor with a PhoB promoter and a GFP reporter.

BBa_K2447000 consists of an inducible promoter (BBa_K116401) that is activated by the binding of phosphorylated PhoB transcription factor. The natural genes of the Pho Regulon signaling pathway are replaced with a GFP reporter (BBa_E0040) so the activation of the promoter following low levels of extracellular phosphate would result in GFP expression.

BBa_K1682012

PhoA Promoter Sequence

The PhoA Promoter is a basic, regulatory part created by 2015 HKUST-Rice iGEM. The inducible promoter was originally made to control the expression of the PhoA alkaline phosphatase gene after activation by the phosphorylated PhoB transcription factor.

BBa_K3725110

PhoA Promoter with GFP Reporter

BBa_K3725110, a composite part, is an improvement upon BBa_K2447000 composed of a PhoA promoter and a GFP reporter. The inducible promoter BBa_K1682012, created by 2015 HKUST-Rice iGEM, was originally made to control the expression of the PhoA alkaline phosphatase gene after activation by the phosphorylated PhoB transcription factor. By replacing the downstream PhoA gene with GFP, the part also expresses fluorescence in the presence of low levels of extracellular phosphate.

BBa_K3725130

PhoB Mutated Consensus Promoter

The PhoB Mutated Consensus Promoter is a basic, regulatory part created by Lambert iGEM. The promoter utilizes a PhoB consensus sequence that mutated the PhoB recognition/binding site of the BBa_K116401 promoter.

BBa_K3725120

PhoB Mutated Consensus Promoter with GFP Reporter

BBa_K3725120 is a composite part containing a mutated promoter that utilizes a PhoB consensus sequence. We engineered the promoter BBa_K3725130 by replacing parts of the PhoB promoter sequence with the PhoB recognition/binding consensus sequence found in literature.

BBa_K3725140

PhoB Redesigned Mutated Consensus Promoter

The PhoB Mutated Consensus Promoter is a basic, regulatory part created by Lambert iGEM. The promoter utilizes a potentially more efficient consensus sequence by directly mutating the RNS polymerase binding site.

BBa_K3725150

PhoB Redesigned Mutated Consensus Promoter w/ GFP Reporter

In an attempt to increase the sensitivity of BBa_3725130 to extracellular phosphate, we designed the promoter BBa_K3725140. The new composite part BBa_K3725150 includes the modified promoter, serving as the phosphate-sensitive biosensor.

NAR PARTS DESCRIPTION

K3725210

BBa_I746909 Superfolder GFP driven by T7 promoter

BBa_I746909 is a generator T7 promoter that drives sfGFP expression.

K3725210

Phosphorylated NarL-Specific Promoter Sequence (pNar)

BBa_K3725210 is a regulatory promoter responsive to phosphorylated NarL. It was designed by Adam Silverman from Jewett Lab at Northwestern University.

K3725220

Phosphorylated NarL-Specific w/ GFP Reporter

BBa_K3725210 is a composite part including the pNar promoter BBa_K3725210, sfGFP, and trrnb terminator. In the presence of phosphorylated NarL, pNar initiates the transcription and translation of downstream sfGFP.

PPB PARTS DESCRIPTION

K3725050

Phytophthora Toehold Switch Sequence

The Phytophthora Toehold Switch Sequence is a basic, intermediate part created by Lambert_GA 2021, and is used within the novel BBa_K3725010 Phytophthora Toehold w/ GFP Reporter.

This Phytophthora Toehold Switch Sequence contains a stem-loop domain and a trigger-complementary region. The trigger-complementary region is derived from the X24 gene, which is required for pathogenicity in Phytophthora cryptogea. We designed this toehold switch using Takahashi et. al.'s Nucleic Acid Package (NUPACK) software code provided by Mrs. McSweeney from the Styzcynski Lab at Georgia Tech.

K3725030

Phytophthora Trigger Sequence

The Phytophthora Trigger Sequence is a basic, intermediate part created by Lambert_GA 2021.

Used in conjunction with the T7 promoter, The Phytophthora trigger RNA sequence can be used to induce GFP expression for Lambert iGEM's Phytophthora w/ GFP reporter. When the trigger RNA is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop, allowing the reporter protein, GFP, to be expressed. This complementary sequence was generated using Nucleic Acid Package (NUPACK) software, and the trigger was experimentally tested with the part BBa_K3725010 for fluorescence.

K3725010

Phytophthora Toehold w/ GFP reporter

BBa_K3725010 Phytophthora Toehold w/ GFP reporter is a construct that the Lambert iGEM 2021 team developed. This composite construct is composed of a T7 promoter, BBa_J64997, the Toehold sequence, BBa_K3725050, GFP, BBa_E0040, a green fluorescent protein as a reporter, and the T7 terminator, and a T7 terminator, BBa_K731721. This part was assembled into the plasmid, pSB1A3. In the presence of the complementary trigger sequence, this toehold sequence will express GFP.

K3725040

T7 Phytophthora Trigger

BBa_K3725040 T7 Phytophthora Trigger is a construct that the Lambert iGEM 2021 team developed. This composite construct is composed of a T7 promoter, BBa_J64997, the Trigger sequence, BBa_K3725030, and a T7 terminator, BBa_K731721. is part was assembled into the plasmid, pSB1K3. This is an RNA sequence intended to be used in conjunction with the part BBa_K3725010 within a biosensor system. When the trigger RNA is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop, allowing the reporter protein, GFP, to be expressed. This complementary sequence was generated using Nucleic Acid Package (NUPACK) software, and the trigger was experimentally tested with the part BBa_K3725010 for fluorescence.

K3725080

Fusarium Toehold Switch Sequence

The Fusarium Toehold Switch Sequence is a basic, intermediate part created by Lambert_GA 2021.

Used in conjunction with the T7 promoter, The Fusarium trigger RNA sequence can be used to induce GFP expression for Lambert iGEM's Fusarium Toehold Switch w/ GFP reporter and Improved FusariumToehold Switch w/ GFP reporter. When the trigger RNA is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop, allowing the reporter protein, GFP, to be expressed. This complementary sequence was generated using Nucleic Acid Package (NUPACK) software, and the trigger was experimentally tested with the part BBa_K3725020 and BBa_K3725022 for fluorescence.

K3725060

Fusarium Trigger Sequence

The (Phytophthora/ Fusarium) trigger is an RNA sequence used in conjunction with the T7 promoter to induce GFP expression for Lambert iGEM's (Phytophthora w/ GFP reporter/ Fusarium Toehold Switch w/ GFP reporter and Improved Fusarium Toehold Switch w/ GFP reporter). When the trigger RNA is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop, allowing the reporter protein, GFP, to be expressed. This complementary sequence was generated using Nucleic Acid Package (NUPACK) software, and the trigger was experimentally tested with the part (BBa_K3725010/ BBa_K3725020 and BBa_K3725022) for fluorescence.

K3725020

Fusarium Toehold w/ GFP reporter

BBa_K3725020 Fusarium Toehold w/ GFP reporter is a construct that the Lambert iGEM 2021 team developed. This composite construct is composed of a T7 promoter, BBa_J64997, the Toehold sequence, BBa_K3725080, GFP, BBa_E0040, a green fluorescent protein as a reporter, and the T7 terminator, and a T7 terminator, BBa_K731721. This part was assembled into the plasmid, pSB1A3. In the presence of the complementary trigger sequence, this toehold sequence will express GFP.

K3725070

T7 Fusarium Trigger

This is the composite of the part, K3725060, a T7 promoter and a T7 terminator. This allows the Fusariumtrigger sequence, which is derived from the FRP1 gene in Fusarium oxysporum, to be constitutively expressed and activate the toehold domain.

BBa_K3725070 T7 Fusarium Trigger is a construct that the Lambert iGEM 2021 team developed. This composite construct is composed of a T7 promoter, BBa_J64997, the Trigger sequence, BBa_K3725060, and a T7 terminator, BBa_K731721. is part was assembled into the plasmid, pSB1K3. This is an RNA sequence intended to be used in conjunction with the part BBa_K3725020 and BBa_K3725022, within a biosensor system. When the trigger RNA is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop, allowing the reporter protein, GFP, to be expressed. This complementary sequence was generated using Nucleic Acid Package (NUPACK) software, and the trigger was experimentally tested with the part BBa_K3725020 and BBa_K3725022 for fluorescence.

K3725082

Improved Fusarium Toehold Switch Sequence

The Fusarium Toehold Switch Sequence is a basic, intermediate part created by Lambert_GA 2021, and is used within the novel BBa_K3725022 Improved Fusarium Toehold w/ GFP Reporter.

This Fusarium Toehold Switch Sequence contains a stem-loop domain and a trigger-complementary region. The trigger-complementary region is derived from the FRP1 gene, which is required for pathogenicity in Fusarium oxysporum f. sp. lycopersici. We designed this toehold switch using Takahashi et. al.'s Nucleic Acid Package (NUPACK) software code provided by Mrs. McSweeney from the Styzcynski Lab at Georgia Tech.

K3725022

Improved Fusarium Toehold w/ GFP reporter

BBa_K3725022 Improved Fusarium Toehold w/ GFP reporter is a construct that the Lambert iGEM 2021 team developed. This composite construct is composed of a T7 promoter, BBa_J64997, the Toehold sequence, BBa_K3725082, GFP, BBa_E0040, a green fluorescent protein as a reporter, and the T7 terminator, and a T7 terminator, BBa_K731721. This part was assembled into the plasmid, pSB1A3. In the presence of the complementary trigger sequence, this toehold sequence will express GFP.