Temporary

Part:BBa_J70572:Design

Designed by: Julie Norville   Group: Knight Lab   (2010-04-30)

J70570(AK) +J70568(AT) into (AC)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1671
    Illegal AgeI site found at 1783
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 836


Design Notes

original J70570 and J70568 had bad restriction enzyme sites in them

which were J70561 and J70550

Source

designed

digest J70562-AK with BsaXI and insert this to add a C-terminal tag

forward (J70562-fi): aaGGATCCCACCACCACCACCACCACTAATAAtactagtagcggc

reverse (J70562-ri): gctactagtaTTATTAGTGGTGGTGGTGGTGGTGGGATCCtttgt

look for green colonies, send six for sequencing, colony pcr-- think about doing his-tag purification later

References