Coding

Part:BBa_M10024:Design

Designed by: Hank Shih   Group: Berkeley BioE140L - S09   (2009-05-09)
Revision as of 02:25, 10 May 2009 by Hshih21 (Talk | contribs) (→‎Design Notes)

{a~Int-native>} Intimin (native)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 405
    Illegal PstI site found at 653
    Illegal PstI site found at 1100
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 405
    Illegal PstI site found at 653
    Illegal PstI site found at 1100
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 405
    Illegal PstI site found at 653
    Illegal PstI site found at 1100
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 405
    Illegal PstI site found at 653
    Illegal PstI site found at 1100
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1248


Design Notes

Construction of {a~Inv-native>} basic part
PCR Bhs001F/Bhs002R on E. coli strain 0157:H7     (146 bp, gp = A)
PCR Bhs002F/Bhs003R on E. coli strain 0157:H7     (1889 bp, gp = B)
----------------------------
PCR Bhs001F/Bhs003R on A+B                       (2009 bp, EcoRI/BamHI)
Digest pBca9495CA-Bca1144#5                      (EcoRI/BamHI, 3039+910, L)
Product is pBca9495CA-M10024                     {a~Inv-native>}
----------------------------
Bhs001F  Forward EcoRI for {a~Inv-native>}          cccaaGAATTCatgAGATCTtaacATGATTACTCATGGTTG
Bhs002F  Removing the EcoRI site from {a~Inv-native>}   GTTAATCAGAACTCATTTGCAAATGG
Bhs002R  Removing the EcoRI site from {a~Inv-native>}  CCATTTGCAAATGAGTTCTGATTAAC
Bhs003R  Reverse BamHI for {a~Inv-native>}          GCAAAggatccGGCCTTGGTTTGATCAAAAAATATAACCGCAC

Source

Genomic sequence of E. coli strain 0157:H7

References