Part:BBa_K3021005

pilA-Laccase-HisTag

This is a new composite part designed to allow Laccase secretion from our engineered cyanobacteria through embedded the secretion peptide, pilA1. Laccase is an important enzyme, which would degrade different chemicals. The Laccase sequence in this part is based on the laccase cDNA sequence from Trametes versicolor in iGEM part k863030. Secretion of heterologous proteins into cell culture medium offers significant advantages for downstream processing over production. pilA1 is the commonly know with its effect on the cyanobacteria secretion.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 928
  Illegal BglII site found at 1360
  Illegal XhoI site found at 960
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1599
  Illegal AgeI site found at 327
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 882
  Illegal SapI.rc site found at 392
  

Sequencing and sequence alignment of K3021005

< img src="https://2019.igem.org/wiki/images/5/56/T--PuiChing_Macau--Alignment_1.jpg">

Figure 3. Representative sequencing results of K3021005_pSCBe (pilA_K863030_pSCBe)

To future confirm that our constructs have correct sequence, we send our clones (with positive PCR results) out to sequencing.

The sequencing results show that we have successfully assembled target genes K3021005 (pilA_K863030_6xHis) to the vector (pSCBe).