Part:BBa_K4032102

lacI+RBS+α-gal+double terminator

Contents:

・The lac promoter (from BBa_R0010)

・The RBS (from BBa_B0034)

・The E.coli alpha-galactosidase gene melA （BBa_K4032000）

・The double terminator (from BBa_B0015)

Usage and Biology

This enzyme catalyzes the hydrolysis of terminal, non-reducing alpha-D-galactose residues in alpha-D-galactosides, including galactose oligosaccharides, galactomannans and galactolipids. https://www.uniprot.org/uniprot/P06720

Design

The lac promoter and the double terminator are added to BBa_K4032005, forming this part.

The lac promoter and the double terminator are used from BBa_J04450.

This part was created by In-Fusion method using BBa_J04450 and melA from E.coli K-12 as an insert.

Fig.1 The plasmid design of K40320102

Experiments

Time course BL21(DE3)

Fig. 2 The growth of E.coli (BL21(DE3)) expressing of alpha-galactosidase

Pre-culture : 37 ℃, 16 h (130 rpm)

Culture : 37 ℃ (130rpm)

･Measuring OD600 every 4 hours

DH5α

Fig. 3 The growth of E.coli (DH5α)) expressing of alpha-galactosidase

Pre-culture : 34 ℃, 16 h (130 rpm)

Culture : 37 ℃ (130rpm)

･Measuring OD600 every 4 hours

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 2137
  Illegal AgeI site found at 2249
  
• 1000
  COMPATIBLE WITH RFC[1000]