DNA

Part:BBa_K676007:Design

Designed by: Kheng Tee Ng   Group: iGEM11_UCL_London   (2011-09-17)

Gyrase A


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1593
    Illegal BglII site found at 2538
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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

References