Part:BBa_K3272005

LTNF without stop codon in pPIC9K

The original coding sequence of the LTNF is inserted in a pPIC9K vector to express it in Pichia pastoris. The DNA sequence was optimized for use in Pichia pastoris by using IDT’s Codon Optimization tool. In this construct, we excluded stop codon before the histidine cleavage site of the vector. By this way, we gain our protein with histidine tag, which will make easier to detect and purify our protein. This construct is used to compare the efficiency of His-tagged LTNF to His-tagged LTNF with two Cysteine residues. The vector contains AOX1 promoter for methanol-induced expression, α-factor secretion signal to direct secreted expression, kanamycin and ampicillin resistance gene for selectivity, the c-myc epitope and polyhistidine (6xHis) tag for detection and purification of our protein.

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]