Part:BBa_K596004

Algae Protein Expression Vector

This Biobrick Plasmid is created by putting biobrick prefix and suffix between HSP70/RBSC2 promoter and 3'UTR. We used pRbcRL(Hsp196)for backbone plasmid. We excised luciferase gene(crluc)using XhoI/BamHI restriction. Then we ligated with biobrick prefix and suffix. This plasmid can be used for protein expression in C.Reinhardtii. HSP70 is a transcriptional activator that enhances expression of a gene.

References: 1. Schroda M, Blöcker D, Beck CF. The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The Plant journal : for cell and molecular biology. 2000;21(2):121-31. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10743653.

2. Fuhrmann M, Hausherr A, Ferbitz L, et al. Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene. Plant molecular biology. 2004;55(6):869-81. Available at: http://www.ncbi.nlm.nih.gov/pubmed/15604722.

Map:

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 3787
  Illegal suffix found in sequence at 1
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3787
  Illegal NheI site found at 3376
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3793
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3787
  Illegal BamHI site found at 23
  Illegal XhoI site found at 3779
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 3787
  Illegal suffix found in sequence at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 3787
  Illegal XbaI site found at 3802
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 2598
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1621
  Illegal SapI site found at 538