Part:BBa_K4288005

cadA promoter-signal peptide of RpmG-ferulyol esterase

cadA promoter-signal peptide of RpmG-ferulyol esterase

BBa_K4288005

Name: cadA promoter-signal peptide of RpmG-feruloyl esterase

Base Pairs: 1128 bp

Origin: Bacillus subtilis, genome

Properties: response to the cadmium concentration

Usage and biology

The biosensor used to monitor the cadmium and zinc in the environment.

BBa_K4288000

Name: cadA promoter

Base Pairs: 159 bp

Origin: Bacillus subtilis, 50S ribosomal protein

Properties: signal peptide

Usage and biology

The cadmium resistance in bacteria using Cad A system. in high concentration of cadmium, the cad A system uptake the cadmium by ATP-dependent pump.

BBa_K4288002

Name: signal peptide of RpmG

Base Pairs: 87 bp

Origin: Bacillus subtilis, 50S ribosomal protein

Properties: signal peptide

Usage and biology

BBa_K4288002 is a coding sequence of RpmG, which is an amino acid sequence of 1-29. we used it as signal peptide.

BBa_K4288001

Name: Feruloyl esterase

Base Pairs: 870 bp

Origin: E.coli

Properties: hydrolysis the ester bands

Usage and biology

Feruloyl esterase (FAE) produced from bacteria, fungi and plant. The enzyme mainly functions as hydrolysis the ester bands. based on amino acids sequence alignment and substrate specificities, the feruloyl esterase classified into four types, including A, B, C, and D. The type D of feruloyl esterase acts as xylanase, which is the hemicellulolytic enzyme. thus, the feruloyl esterase was used as additive to improve the bioconversion and utilization of the agricultural stuff.

Construct design

The reporter feruloyl esterase gene controlled under the cad promoter which monitors the cadmium and zine concentration of solution. Further, the reporter fused the signal peptide RpmG. The gene fragment was inserted into the pHY300PLK vector.

Experimental approach

We synthesized the complete gene fragment including cadA promoter, signal peptide RpmG, and biosensor feruloyl esterase. The gene was further cloned in pHY300PLK by BamHI and EcoR1 (Figure 1).

Figure 1. The map of pHY300PLK cadA-RpmG-feruloyl esterase.

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]