Difference between revisions of "Part:BBa K1898250:Design"

Revision as of 03:57, 19 October 2016

Strong promoter + Strong RBS + GSR + 10x Histidine tag + Double terminator

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 804
Illegal BamHI site found at 1488
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 89
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 1199
Illegal SapI.rc site found at 1607

Design Notes

We did five silent point mutations to the GSR sequence to remove the two Pst1 and three EcoR1 internal cutting sites. The sequence was sent out for mutagenesis.

Primers were designed to remove the stop codon from GSR and to move the cDNA into iGEM Biobrick:

forward: 5' ATATgAATTCgCggCCg 3' (17)

reverse: 5' ATATCTgCAgCggCC 3' (15)

Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. This is showed in this part's main page under sequencing. The sequence was sent to mutagenesis to add two base pairs in between the two genes.

Source

the cDNA of GSR was ordered from OriGene. Mutagensis were performed by MissionBiotech. Primers were synthesized by Tri-I Biotech.

@@ Line 9: / Line 9: @@
 We did five silent point mutations to the GSR sequence to remove the two Pst1 and three EcoR1 internal cutting sites. The sequence was sent out for mutagenesis.
-Primers were designed to remove the stop codon from GSR and to move the cDNA into iGEM Biobrick.
+Primers were designed to remove the stop codon from GSR and to move the cDNA into iGEM Biobrick:
+forward: 5' ATATgAATTCgCggCCg 3' (17)
+reverse: 5' ATATCTgCAgCggCC 3' (15)
 Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. This is showed in this part's main page under sequencing.