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 （）

・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 No part name specified with partinfo tag., 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 K40320xx

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]