https://pages.htgaa.org/2026b/francis-chanda/index.htmlAbout me I’m a Bioinformatician from Zambia with a background in biology, bioinformatics and data science. I enjoy building things and figuring out how technology can be used to solve practical problems, especially in biology and healthcare.Hugoenhttps://pages.htgaa.org/2026b/francis-chanda/week-01-hw-project-ideas/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/francis-chanda/week-01-hw-project-ideas/index.htmlBiological −20°C Low-Cost Fluorescent Time–Temperature Indicator Sticker What I would like to biomanufacture and why? What is a Biological −20°C Low-Cost Fluorescent Time–Temperature Indicator Sticker? This a sticker that can be attached to temperature-sensitive laboratory reagents and provide an early visual warning when they experience inappropriate warming. The indicator would ideally respond to both the temperature reached and the duration of exposure. Under appropriate storage conditions, it would remain in a low-fluorescence state. When the temperature rises above the −20°C storage threshold, it would begin producing a detectable fluorescent signal. A longer or more severe temperature excursion would produce a stronger signal, providing an indication of the severity of the exposure.https://pages.htgaa.org/2026b/francis-chanda/week-02-hw-dna-design/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/francis-chanda/week-02-hw-dna-design/index.htmlRepository : Chanda-BiologicalFTTI Expressing : GFP-ssrA and AmilCP GFP-ssrA iGEM Registry -> BBa_K819007 5’ - CTTGTGGCAACAATTTCTACAAACACTTGATACCGTATGAGCATACGGTATAATTGCTTCAACAGAACATATTGACTATCATTAAAGAGGAGAAATACTAGATGCGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATT AGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGGTTATGGTGTTCAAT GCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTA AGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTT GTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGCTGCTAACGACGAAAACTACGCTCTGGCTGCTTACTAGTAG - ‘3 AmilCP iGEM Registry -> BBa_K3988019 5’ - ATGTCAGTGATCGCTAAGCAAATGACATACAAAGTCTATATGTCTGGGACCGTAAATGGCCACTACTTTGAAGTGGAAGGGGATGGCAAGGGGAAGCCTTACGAGGGTGAACAGACCGTAAAACTGACTGTAACAAAAGGGGGCCCTTTACCTT TCGCATGGGACATCCTTTCCCCTCAATGTCAGTACGGCTCCATCCCTTTTACGAAGTACCCGGAGGATATCCCTGATTATGTTAAGCAGTCTTTCCCAGAAGGCTACACCTGGGAACGGATTATGAATTTCGAGGATGGTGCGGTTTGTACCGTCTCAAAC GACTCATCGATCCAAGGTAACTGCTTCATTTATCACGTTAAATTTTCTGGTCTCAATTTTCCACCTAATGGTCCGGTCATGCAGAAAAAAACCCAAGGGTGGGAGCCAAACACGGAGCGGTTGTTCGCACGCGACGGCATGTTACTCGGGAACAATT Host Chasis : E.Coli Layer Part Type Source Justification Sensor C120 Promoter Kernel Provides continuous expression of the engineered thermal transcriptional regulator so it is available to respond when temperature changes. Sensor BBa_B0034 RBS Kernel Enables efficient translation of the engineered AsLOV2-EL222 HTH fusion in E. coli. Sensor Engineered AsLOV2-I532A + EL222 HTH Domain CDS Designed Combines the engineered temperature-responsive AsLOV2-I532A module with the EL222 DNA-binding domain to couple temperature-dependent conformational changes to transcriptional regulation. Sensor Terminator Terminator Kernel Real-Time Output C120 Promoter Kernel Provides the promoter activated by the EL222 HTH domain to drive the temperature-responsive reporter. Real-Time Output BBa_B0034 RBS Kernel Enables translation of the fluorescent reporter in E. coli. Real-Time Output GFP-ssrA CDS Kernel Provides a fluorescent signal while the thermal response is active, with the ssrA tag promoting degradation after the signal is switched off. Real-Time Output Terminator Terminator Kernel Terminates transcription of the GFP reporter cassette. Memory C120 Promoter Same as output Memory BBa_B0034 RBS Kernel Provides a well-characterized ribosome-binding site for efficient translation initiation in E. coli. Memory amilCP CDS Kernel Uses the same thermal-responsive regulatory input so that cumulative colour production begins under the same temperature conditions as the fluorescent response. Memory Terminator Terminator Kernel Provides reliable transcription termination, helping define the end of each expression cassette and reduce unwanted read-through transcription. Vector E.Coli Backbone Backbone Existing vector Provides the replication origin and selectable marker required to maintain the construct in E. coli. Sequences before Fusing both to create Chimeric Protein Engineered AsLOV2-I532A 5’ GEFLATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGRNCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQYFIGVQLDGTEHVRDAAEREGVMLAKKTAENIDEAAKEL ‘3 EL222 HTH Domain 5’ ATGTTGGATATGGGACAAGATCGGCCGATCGATGGAAGTGGGGCACCCGGGGCAGACGACACACGCGTTGAGGTGCAACCGCCGGCGCAGTGGGTCCTCGACCTGATCGAGGCCAGCCCGATCGCATCGGTCGTGTCCGATCCGCGTCTCGCCGACAATCCGCTGATCGCCATCAACCAGGCCTTCACCGACCTGACCGGCTATTCCGAAGAAGAGTGCGTCGGCCGCAATTGCCGATTCCTGGCAGGTTCCGGCACCGAGCCGTGGCTGACCGACAAGATCCGCCAAGGCGTGCGCGAGCACAAGCCGGTGCTGGTCGAGATCCTGAACTACAAGAAGGACGGCACGCCGTTCCGCAACGCCGTGCTCGTTGCACCGATCTACGACGACGACGACGAGCTTCTCTATTTCCTCGGCAGCCAGGTCGAAGTCGACGACGACCAGCCCAACATGGGCATGGCGCGCCGCGAACGCGCCGCGGAAATGCTCAAGACGCTGTCGCCGCGCCAGCTCGAGGTTACGACGCTGGTGGCATCGGGCTTGCGCAACAAGGAAGTGGCGGCCCGGCTCGGCCTGTCGGAGAAAACCGTCAAGATGCACCGCGGGCTGGTGATGGAAAAGCTCAACCTGAAGACCAGCGCCGATCTGGTGCGAATTGCCGTCGAAGCCGGAATCTGA ‘3’https://pages.htgaa.org/2026b/francis-chanda/week-03-hw-cell-free/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/francis-chanda/week-03-hw-cell-free/index.htmlA. COMPARE CELL-FREE REACTION COMPOSITIONS (20 minutes) 1. Why does the concentration of potassium glutamate and magnesium glutamate differ between compositions? Which is more sensitive? Magnesium Glutamate is more sensitive when compared to Potassium Glutamate, small deviations from the optimal concentration can lead to significant drops in protein production [20] where as research has shown that protein production is relatively independent of potassium glutamate concentration over a wide range [20]. Magnesium (Mg²⁺) binds to ATP and other nucleotides, and phosphate can pull it out of solution. So a recipe with more nucleotides or phosphate needs more Mg²⁺ to keep enough free for the ribosomes and T7 RNA polymerase. Potassium (K⁺) mostly sets the overall ionic strength and helps ribosomes and translation factors work, so it can vary over a wide range, often roughly 80-200 mM.https://pages.htgaa.org/2026b/francis-chanda/final-project/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/francis-chanda/final-project/index.html