Composite

Part:BBa_J70310:Design

Designed by: Tom Knight   Group: Knight Lab   (2009-06-16)
Revision as of 22:52, 21 July 2009 by Tk (Talk | contribs) (→‎Source)

Mesoplasma florum pBG7AU plasmid origin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 99
    Illegal NheI site found at 120
    Illegal NheI site found at 141
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 779
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 747
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Unclear location for the cut site of the circular plasmid.



Source

PCR from recircularized pBG7AU cut at the ClaI site. Original plasmid from Djordjevic01. Primers:

GTTTCTTC GAATTC GCGGCCGC T TCTAGA Gctcctttttttggggttagcgatag,BG7AU-F2

GTTTCTTC CTGCAG CGGCCGC T ACTAGT Acaaaaaaaggagaatttactcc,BG7AU-R


Original part design used this primer, which had primer-dimer issues with -R primer. Redesign using -F2 primer, modified the part sequence at the 5' end. GTTTCTTC GAATTC GCGGCCGC T TCTAGA Ggagtaaattctcctttttttgg,BG7AU-F

References