Part:BBa_K528020

Expression Adapter s03_65k

This part encodes expression adaptor designed to have strength 92,86% relative to B0034 and uniform expression level regardless of attached protein.

Authors

Clonning:

• Paweł Urbański
• Elżbieta Jankowska
• Krzysztof Szczepaniak
• Dorota Kaja Sabat

The lab work was supervised by:

• Michał Lower
• Ania Olchowik

Construct design

The part is in RFC25 standard. It consists of BBa_J23100 promoter followed by BBa_B0034 RBS, 5 nucleotide spacer and short peptide.

Spacer and peptide were designed using in silico evolution to provide uniform expression level regardless of following protein coding sequence.

Summary of design algorithm:

Measurements

Using RBSCalculator program we have determined that this part expression strength is 92,86% relative to BBa_B0034

All our measurement results are summarized in the table below:

Part Name	Description	Expression level in thousands of absolute RBSDesigner units	Expression level as %B0034+YFP
BBa_K528015	Expression Adapters s00_45k	45	64,29%
BBa_K528016	Expression Adapters s01_24k	24	34,29%
BBa_K528017	expression adapters s02_20k	20	28,57%
BBa_K528018	Expression Adapters s06_16k	16	22,86%
BBa_K528019	Expression Adapters s05_12k	12	17,14%
BBa_K528020	expression adapters s03_65k	65	92,86%
BBa_K528021	BBa_K528018+YFP measurement device	16	22,86%
reference	BBa_J23102+B0034+YFP (100% reference construct)	70	100,00%

For more info please look at Team Warsaw 2011 [http://2011.igem.org/Team:Warsaw/ExpressionAdaptors wiki page].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 8
  Illegal NheI site found at 31
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]