Part:BBa_K1717171
Protein coding sequence for KeratinaseA
Keratinases are proteolytic enzymes which break the disulphide bonds in various keratin substrates. KERA is a basic part comprised from a synthesized KERA protein coding sequence, optimized for expression in E. coli and most active in degrading keratin in feathers. This gene, when expressed in cells, can be tested for activity through looking for zones of clearing on skim milk agar plates where cells are growing, as well as by looking at degradation of keratin-containing substrates (feathers, hair, chemically synthesized keratin, etc.) when grown with cells.
SZ_SHD 2021's Improvement
Introduction
The composite part BBa_K1717171 is an optimized version of KERA that can be constructed into pSB1C3 plasmid with the highest protein expression in E.coli BL21(DE3). The KERA is a basic part comprised from a synthesized KERA protein coding sequence. The gene can be expressed in cells by looking for clear areas of cell growth on AGAR plates of skimmed milk, and by observing keratin-containing substrates (feathers, hair, chemosynthetic keratin, etc.) growing with cells.
Plasmid Construction
Experiment details
• Throughput: 2500
• Organism: E. coli
• Strain type: BL21(DE3)
• Temperature: 37°C
• Media: 250 ml LB
• Period of time: 16 hours
• Plasmid backbone: pSB1C3
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 130
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 391
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 761
Illegal SapI site found at 1064
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