Team:Lambert GA/Parts

Parts Overview

In conjunction with the 2019 project to detect helminths in fecal and soil samples, Lambert iGEM submitted seven parts to the Registry of Standard Biological Parts, four of which are specific to the Caenorhabditis elegans lin-4 gene pre-mRNA primary transcript F59G1.6. T7 C. elegans trigger (BBa_K2974400) is an RNA sequence to be used with BBa_J23106 C. elegans Toehold GFP (BBa_K2974316) to induce GFP expression in a biosensor system. When the trigger RNA sequence is present, it will bind to the complementary sequence in the toehold switch and unbind the hairpin loop allowing the reporter protein to be expressed.

Two of the parts, BBa_K2974101 and BBa_K2974700, were designed using Lambert 2018's toehold sequence and GFP in order to draw a comparison between the effects of weak and strong promoters within riboregulators. Last year, a strong T7 promoter had the unintended side effect of overexpression and induced downstream transcription without the presence of the trigger sequence, leading to false-positive results. In compliance with iGEM's gold medal requirement for improvement, BBa_K2974500 was submitted with the same Cholera-specific toehold switch and LacZ reporter but with a weaker promoter to combat overexpression.

New Parts

♡ = Favorite Part
✓ = Confirmed Sequencing

Work Stream Name Type Description Designer Length
✓ C. elegans BBa_K2974410 Intermediate C. elegans Trigger Megan Hong, Courtney Taing, Julia Chang, Sri Pinnamareddy, Shanthi Hegde, Janet Standeven 100
♡ ✓ C. elegans BBa_K2974310 Intermediate C. elegans Toehold Switch Megan Hong, Courtney Taing, Julia Chang, Sri Pinnamareddy, Shanthi Hegde, Janet Standeven 471
♡ ✓ C. elegans BBa_K2974316 Composite BBa_J23106 C. elegans Toehold GFP Megan Hong, Courtney Taing, Julia Chang, Sri Pinnamareddy, Shanthi Hegde, Janet Standeven 3168
✓ C. elegans BBa_K2974400 Composite T7 C. elegans Trigger Megan Hong, Courtney Taing, Julia Chang, Sri Pinnamareddy, Shanthi Hegde, Janet Standeven 2585
✓ C. elegans Proof of Concept BBa_K2974101 Composite BBa_J23106 Toehold GFP Abby Bell, Ashtyn Cauffman, Monica Cho, Janet Standeven 5347
✓ C. elegans Proof of Concept BBa_K2974700 Composite BBa_J23100 Toehold GFP Abby Bell, Ashtyn Cauffman, Monica Cho, Janet Standeven 3977
Improved BBa_K2974500 Composite BBa_J23106 Toehold LacZ Abby Bell, Ashtyn Cauffman, Monica Cho, Janet Standeven 3977

CHARACTERIZED PARTS

Work Stream Name Type Description Designer Length
✓ Improved BBa_K2550000 Composite BBa_J23100 Toehold Ribosome Switch with LacZ expression Janet Standeven, Abby Bell, Megan Hong, Yan Zhang, Courtney Taing, Ashtyn Cauffman 3218
✓ Improved BBa_K2550001 Composite T7 promoter with a trigger sequence to match the toehold sequence of part BBa_K2550000 Yan Zhang, Janet Standeven, Abby Bell, Megan Hong 152
✓ Modeling BBa_J10050 Composite Weak promoter + Weak RBS + LacZa.GFP Fusion Natalie Kuldell 1017
✓ Modeling BBa_J10051 Composite Weak promoter + Strong RBS + LacZa.GFP Fusion Natalie Kuldell 1015
✓ Modeling BBa_J10052 Composite Weak promoter + Medium RBS + LacZa.GFP Fusion Natalie Kuldell 1016
✓ Modeling BBa_J10053 Composite Medium promoter + Weak RBS + LacZa.GFP Fusion Natalie Kuldell 1017
✓ Modeling BBa_J10054 Composite Medium promoter + Medium RBS + LacZa.GFP Fusion Natalie Kuldell 1016
✓ Modeling BBa_J10055 Composite Medium promoter + Strong RBS + LacZa.GFP Fusion Natalie Kuldell 1015
✓ Modeling BBa_J10056 Composite Strong promoter + Weak RBS + LacZa.GFP Fusion Natalie Kuldell 1017
✓ Modeling BBa_J10057 Composite Strong promoter + Medium RBS + LacZa.GFP Fusion Natalie Kuldell 1016
✓ Modeling BBa_J10058 Composite Strong promoter + Strong RBS + LacZa.GFP Fusion Natalie Kuldell 1015
✓ Modeling BBa_J23115 Promoter Constitutive Promoter Family Member John Anderson 35
✓ Modeling BBa_B0035 Ribosomal Binding Site RBS (B0030 derivative) Jason Kelly 14