Difference between revisions of "Part:BBa K1898200:Design"

 
 
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===Design Notes===
 
===Design Notes===
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The purchased GSR cDNA has two internal PstI and three EcoRI cutting sites. After making silent mutations to the sequence, we sent the cDNA to MissionBiotech for mutagenesis to remove these cutting sites
+
The purchased GSR cDNA has two internal PstI and three EcoRI cutting sites. After making silent mutations to the sequence, we sent the cDNA for mutagenesis to remove these cutting sites.
  
 +
Primers were designed to remove the stop codon and to move the DNA into iGEM BioBrick. The primers are show below:
 +
 +
forward: 5' ATATgAATTCgCggCCgCTTCTAgATggCCCTgCTgCCC 3' (39)
 +
 +
reverse: 5' ATATCTgCAgCggCCgCTACTAgTAACgAAgTgTgACCAgC 3' (41)
  
  
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the cDNA of GSR is ordered from Origene.  
 
the cDNA of GSR is ordered from Origene.  
  
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===References===
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Mutagenesis is performed by MissionBiotech and primers were synthesized by Tri-I Biotech

Latest revision as of 12:12, 18 October 2016


GSR, glutathione reductase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 743
    Illegal BamHI site found at 1427
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 28
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1138
    Illegal SapI.rc site found at 1546


Design Notes

The purchased GSR cDNA has two internal PstI and three EcoRI cutting sites. After making silent mutations to the sequence, we sent the cDNA for mutagenesis to remove these cutting sites.

Primers were designed to remove the stop codon and to move the DNA into iGEM BioBrick. The primers are show below:

forward: 5' ATATgAATTCgCggCCgCTTCTAgATggCCCTgCTgCCC 3' (39)

reverse: 5' ATATCTgCAgCggCCgCTACTAgTAACgAAgTgTgACCAgC 3' (41)


Source

the cDNA of GSR is ordered from Origene.

Mutagenesis is performed by MissionBiotech and primers were synthesized by Tri-I Biotech