https://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’Hugoen