Part:BBa_K2384013

Repressor gene xylR+Strong PxylA promoter+RBS+GST-CRS5+His-tag

Xylose-inducible expression system: strong PxylA promoter and repressor gene xylR; after xylose addition, the repressor is released from the PxylA that activates transcription initiation.(BBa_K733002) They have been designed for high expression in B. megaterium. This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell(BBa_K1460002). This year our parts were doing codon optimisation but without terminator codon.

Usage and Biology

Optimization Report

1.Codon Used Adjustment The best value is 1 for sequence optimization.

Figure 1:After Codon Adjustment

2.GC Content: The comparison of GC content between original sequence and optimized sequence

Figure 2:After Optimization

Figure 3:The expression of GST-CRS5 detected using SDS-PAGE

Figure 4:Plasmid digested by EcoRI and HindIII

Figure 5:Our genetic circuit

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2159
  Illegal XhoI site found at 847
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1571