Part:BBa_K2273103

AmyE_EV version of B. subtilis alpha-amylase without its native signal peptide

The AmyE_EV which was amplified from AmyE_RFC25 (BBa_K2273104) is used as gene of interest for proof of concept of the Signal Peptide Toolbox of [http://2017.igem.org/Team:TU_Dresden iGEM Team TU Dresden 2017 (EncaBcillus - It's a trap!)].

The AmyE_EV version of B. subtilis alpha-amylase (Uniprot [http://www.uniprot.org/uniprot/P00691 P00691]) without its first 33 amino acids (Brockmeier et al, 2006) which code for the native signal peptide would naturally be secreted into the supernatant to degrade extracellular starch. However, as its native signal peptide has been cut off, it lacks the naturally occuring secretion capability.

This part was generated in a modified version of RFC25 and has the following prefix and suffix:

Prefix with	EcoRI, NotI, XbaI, AgeI and NgoMIV	5'-GAATTCGCGGCCGCTTCTAGAACCGGTGCCGGC-3'
Suffix with	SpeI, NotI and PstI	5'-TACTAGTAGCGGCCGCTGCAG-3'

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI are red. Additionally, the AgeI and NgoMIV are marked in orange. NgoMIV was underlined for clarity.)

Sequence and Features

The DNA sequence of this part has been verified via sequencing before it was sent in.

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]