Plasmid
APEV

Part:BBa_K596004

Designed by: Ebuzer Kalyoncu   Group: iGEM11_Bilkent_UNAM_Turkey   (2011-09-19)
Revision as of 15:59, 21 September 2011 by Rezube (Talk | contribs)

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: Algae protein expression vector pAPEV.png

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


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