Generator

Part:BBa_J70304:Design

Designed by: Tom Knight   Group: Knight Lab   (2009-05-06)


constitutive cat cassette for Mesoplasma florum


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Test for KpnI sites in the source plasmid.


Source

PCR from pSB1T3-J70026 with primers

GTTTCTTCggtacctttcttcattagaatcaatcc,cat-mut-R

GTTTCTTCggtacctttaggaggcatatcaaatg,cat-mut-F

These primers bind to the source plasmid and remove the left side of the hairpin and add a pair of KpnI sites, which will be cut and religated. I hope to select on Cm plates, since the construct should be Cm resistant.


References