DNA
Part:BBa_K676007:Design
Designed by: Kheng Tee Ng Group: iGEM11_UCL_London (2011-09-17)
Gyrase A
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1593
Illegal BglII site found at 2538 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 27
Illegal AgeI site found at 703
Illegal AgeI site found at 760
Illegal AgeI site found at 1399
Illegal AgeI site found at 2155
Illegal AgeI site found at 2523 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 43
Illegal SapI.rc site found at 51
Illegal SapI.rc site found at 1510
Design Notes
This is the E.coli GyrA ORF with a Pst 1 restriction site at the 1212th bp position. SIte directed mutagenesis should be done to remove the site.
Using Eurofins operon ( Prof Ward had kindly allowed us to use his account), we designed the appropriate primers which include the standard EcoR1 and XbaI restriction sites upstream of the ORFs while another SpeI and PstI restriction sites in the downstream of the ORFs. Here are the primers we used: gyrA (For) TAG TTC GAA TTC TCT AGA ATG AGC GAC CTT GCG AGA GAA AT (41 bp) gyrA (Rev) ATC ATC CTG CAG ACT AGT TTA TTC TTC TTC TGG CTC GTC GTC (42 bp)
Source
TOP10 E.coli genome