Part:BBa_K2384006

Pveg+RBS+CspC+6*HIS+T1 terminator+T7 terminator

This is an improve part.This gene is a cold resistant gene from the Arctic cryophile Polaribacter irgensii.It can largely increase the cold resistance of bacterias.CDS sequence optimization in (BBa_K538004) is better suited for expression in Bacillus megaterium and we add a 6*His tag at the C end.After that, the promoter (BBa_K143012) and RBS (BBa_K090505) were combined.

Optimization Report

Expression System:Bacillus megaterium

Gene Length:189 (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

4.Restriction Enzyme

Figure 7:After Optimization

Figure 8: Gene on pSB1C3 Digested with SpeI

Usage and Biology

Freeze/thaw cycle survival rate of Bacillus megaterium containing an empty vector (red line), a vector carrying CspC gene (blue line).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]