Part:BBa_K2384018

Repressor gene xylR+Strong PxylA promoter+RBS+DsbA-MBP(Pb)+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.The periplasm display protein Dsba-MBP, which binding the lead of the enviroment. Disulfide bond formation protein A (DsbA) is a powerful oxidative protein of the thioredoxin family. DsbA can help protein to fold correctly both in vivo and in vitro 。The C-terminal fusion protein of the maltose-binding protein can be efficiently and rapidly output into the periplasm as a key component of the expression of the metal-binding peptide of the cyclin. This year our parts were doing codon optimisation but without terminator codon.

Usage and Biology

Optimization Report

Gene Name:DSBA-MBP(Pb)

Expression System:Bacillus megaterium

Gene Length:963 (bp)

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

Figure 1:Before Codon Adjustment

Figure 2:After Codon Adjustment

2.Codon Used Distribution Show the relative codon used distribution

Figure 3:Before Optimization

Figure 4:After Optimization

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

Figure 5:Before Optimization

Figure 6:After Optimization

Figure 7:The expression of DsbA+MBP(Pb) detected using SDS-PAGE

Figure 8:Plasmid digested by EcoRI-PstI

Figure 9:Our genetic circuit

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 1668
  Illegal BglII site found at 1680
  Illegal XhoI site found at 847
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]