DNA

Part:BBa_J70032:Design

Designed by: Julie Norville   Group: Knight Lab   (2008-12-12)

A Composite BioScaffold Part (PpiI and PsrI) for Protein Fusions (see Design Page)


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 NgoMIV site found at 111
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 43
    Illegal BsaI.rc site found at 54
    Illegal BsaI.rc site found at 69


Design Notes

Can be used to create protein fusions, see BBF RFC15. (The part that links the two BioScaffold parts can be altered as desired, here a 22 bp placeholder is used that leaves at least 7 bp beyond the internal BioBrick scars.) TACTAGAG and TACAGAT represent sequences for BioBrick scars that are created when two BioBrick parts are fused together. ^s represent restriction enzyme cut sites.

General Function (after cutting with PpiI and PsrI)

XXXXX XXXXX^XXX TACTAGAG  BioScaffold Part BBa_J70032 TACTAGAT XX XXXXX^XXXXX
XXXXX^XXXXX XXX ATGATCAC                              ATGATCTA XX^XXXXX XXXXX

Function when the left and right parts are protein parts (after cutting with PpiI and PsrI)

XXXXX XXXXX^TAA TACTAGAG  BioScaffold Part BBa_J70032 TACTAGAT GX XXXXX^XXXXX
XXXXX^XXXXX ATT ATGATCAC                              ATGATCTA CX^XXXXX XXXXX

Function when the left and right parts are protein parts (after cutting with PpiI and PsrI,) with appropriately positioned BamHI sites.

XXXXG GATCC^TAA TACTAGAG  BioScaffold Part BBa_J70032 TACTAGAT GG GATCC^XXXXX
XXXXC^CTAGG ATT ATGATCAC                              ATGATCTA CC^CTAGG XXXXX


What happens after cutting with MabI and purifying with either gel purification or a Qiagen PCR cleanup spin column

General Case

XXXXX XXXXX^                                                    ^XXXXX
XXXXX^                                                    ^XXXXX XXXXX

Case when left and right parts are a protein part

XXXXX XXXXX^                                                    ^XXXXX
XXXXX^                                                    ^XXXXX XXXXX

Case when left and right parts are protein parts with appropriately positioned BamHI sites

XXXXG GATCC^                                                    ^XXXXX
XXXXC^                                                    ^CTAGG XXXXX

Source

Designed

References