Difference between revisions of "Part:BBa K411101:Design"
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===Design Notes===
 
===Design Notes===
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We used PCR to fuse the protein domains together.
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More details of the fusion can be found on [http://2010.igem.org/Team:NYMU-Taipei/Project/Speedy_reporter/Materials_and_Methods#GFP_fusion_part our wiki page].
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===Source===
 
===Source===
  
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PCR fusion of [[Part:BBa_E0040|GFP]] and [[Part:BBa_K411100|eIF4A]]. More details of the fusion can be found on [http://2010.igem.org/Team:NYMU-Taipei our wiki page].
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PCR fusion of amino acids 1-158 of [[Part:BBa_E0040|GFP]], a GSSGSSGSGS peptide linker, and amino acids 1-215 of [[Part:BBa_K411100|eIF4A]].  
 
These four primers were used to fuse them together:
 
These four primers were used to fuse them together:
 
* GFP_split_A_FP: 5' - gaattcgcggccgcttctagag atgcgtaaaggagaagaacttttc - 3' (46bp)
 
* GFP_split_A_FP: 5' - gaattcgcggccgcttctagag atgcgtaaaggagaagaacttttc - 3' (46bp)

Latest revision as of 17:16, 26 October 2010

GFP_split_A + Linker + eIF4A_split_A


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 741
    Illegal BglII site found at 1037
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1139


Design Notes

We used PCR to fuse the protein domains together. More details of the fusion can be found on [http://2010.igem.org/Team:NYMU-Taipei/Project/Speedy_reporter/Materials_and_Methods#GFP_fusion_part our wiki page].

Source

PCR fusion of amino acids 1-158 of GFP, a GSSGSSGSGS peptide linker, and amino acids 1-215 of eIF4A. These four primers were used to fuse them together:

  • GFP_split_A_FP: 5' - gaattcgcggccgcttctagag atgcgtaaaggagaagaacttttc - 3' (46bp)
  • GFP_split_A_RP: 5' - gctgccgctgctgccgctgctgcc ttgtttgtctgccatgatgtatac - 3' (48bp)
  • eIF4A_split_A_FP: 5' - cggcagcagcggcagcggcagc atggagccggaaggcgtcatcga - 3' (45bp)
  • eIF4A_split_A_RP: 5' - ctgcagcggccgctactagta tca agggtctctcataaatttcttgg - 3' (47bp)

References