Part:BBa_K4770018:Design

NR_Nitrate_Reductase_Level_1

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1517
  Illegal PstI site found at 2781
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 3361
  Illegal PstI site found at 1517
  Illegal PstI site found at 2781
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 3916
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1517
  Illegal PstI site found at 2781
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1517
  Illegal PstI site found at 2781
  Illegal NgoMIV site found at 1406
  Illegal NgoMIV site found at 2000
  Illegal NgoMIV site found at 2615
  Illegal NgoMIV site found at 3732
  Illegal AgeI site found at 1178
  Illegal AgeI site found at 3491
  
• 1000
  COMPATIBLE WITH RFC[1000]

Source of this part

- Original PPSAD sequence: BBa_K4770007

- Original NR sequence: BBa_K4770002

- Original F2A sequence: BBa_K4770012

- Original NanoLuc sequence: BBa_K4770011

- Original TPSAD sequence: BBa_K4770008

Design considerations

We performed Chlamydomonas reinhardtii's domestication for NR sequence. This process includes a reduction of the GC% content to make the sequence suitable for commercial synthesis while maintaining it high enough to fit Chlamydomonas reinhardtii's codon usage. This was done using our optimizing software (see AlgaGenix's wiki for additional information). Moreover, recognition sites for BbsI and BsaI were eliminated, changing said codons with synonymous ones. Besides, we also added an intron to perform what is known as intron-mediated enhancement (IME), which studies show aids with stable high-level expression of a foreign gene (Lumbreras et al., 1998) AlgaGenix’s Level 1s are designed to have the same structure and not having to build different pieces with different positions for each gene.