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New BioBricks

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Cripto-1 Expression System (K1908001)

This part is a semi-composite part that was synthesized by IDT which holds a Saccharomyces cerevisiae Kozak sequence, a coding sequence for the Homo sapien membrane protein Cripto-1, and an ADH1 Terminator region. The coding sequence was codon-optimized for expression in yeast using the proprietary codon optimization program on the Integrated DNA Technologies website. Our design was intended to induce a strong expression of Cripto-1 in yeast to test for membrane localization. We hope that our contribution of a eukaryotic receptor protein to the iGEM Registry is found to be useful and beneficial.

Hepatitis B Antigen Expression System (K1908002)

The Hepatitis B Surface antigen (HBsAg) is a subunit from the surface antigen of the Hepatitis B virus. The blue chromoprotein is added as a visual indicator of the expression of the antigen. The 2A is a short amino acid sequence that is commonly used by picornaviruses to cleave proteins, and will allow the separation of the blue chromaprotein from HBsAg. The yeast secretion tag will allow the antigen to be secreted into the media. The supernatant of the media can be tested for the secretion of the antigen using an HBsAg ELISA Kit. This composite part also contains a yeast ADH1 Terminator. We hope that this part would serve as a dual reporter in more complex genetically engineered systems.

BioBrick Improvements

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fwYellow-amilCP Osmolarity Color Toggle (K1908000)

This system is intended to be a blue-yellow color toggle that expresses fwYellow (K1033910) and amilCP (K592009) dependent on the activation of the OmpC promoter at varied ambient osmolarities. A negative feedback loop inhibits production of fwYellow (regulated by pLac) when the OmpC promoter is active and amilCP/LacI is being produced. Growth in high-osmolarity media is expected to give a blue color while growth in low-osmolarity media is expected to produce a yellow color. Further characterization of the construct is necessary to understand the mathematical relationship between protein expression levels at varied ambient osmolarities.

Parts

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Part Name	Part Description
BBa_K1908000	BY-Toggle
BBa_K1908001	Cripto-1 Coding Sequence
BBa_K1908002	HBsAg Expression Construct