Composite

Part:BBa_K571007:Design

Designed by: Lu-Chu Ke   Group: iGEM11_TzuChiU_Formosa   (2011-10-03)


R0011+acsAB+acsCD+CMCax+Ccp/pSB1C3


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 666
    Illegal EcoRI site found at 2067
    Illegal EcoRI site found at 7892
    Illegal EcoRI site found at 11030
    Illegal PstI site found at 822
    Illegal PstI site found at 1218
    Illegal PstI site found at 2058
    Illegal PstI site found at 4802
    Illegal PstI site found at 4988
    Illegal PstI site found at 5438
    Illegal PstI site found at 7274
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 666
    Illegal EcoRI site found at 2067
    Illegal EcoRI site found at 7892
    Illegal EcoRI site found at 11030
    Illegal PstI site found at 822
    Illegal PstI site found at 1218
    Illegal PstI site found at 2058
    Illegal PstI site found at 4802
    Illegal PstI site found at 4988
    Illegal PstI site found at 5438
    Illegal PstI site found at 7274
    Illegal NotI site found at 3233
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 666
    Illegal EcoRI site found at 2067
    Illegal EcoRI site found at 7892
    Illegal EcoRI site found at 11030
    Illegal BglII site found at 1222
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 666
    Illegal EcoRI site found at 2067
    Illegal EcoRI site found at 7892
    Illegal EcoRI site found at 11030
    Illegal PstI site found at 822
    Illegal PstI site found at 1218
    Illegal PstI site found at 2058
    Illegal PstI site found at 4802
    Illegal PstI site found at 4988
    Illegal PstI site found at 5438
    Illegal PstI site found at 7274
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 666
    Illegal EcoRI site found at 2067
    Illegal EcoRI site found at 7892
    Illegal EcoRI site found at 11030
    Illegal PstI site found at 822
    Illegal PstI site found at 1218
    Illegal PstI site found at 2058
    Illegal PstI site found at 4802
    Illegal PstI site found at 4988
    Illegal PstI site found at 5438
    Illegal PstI site found at 7274
    Illegal NgoMIV site found at 2441
    Illegal NgoMIV site found at 2481
    Illegal NgoMIV site found at 5321
    Illegal NgoMIV site found at 6397
    Illegal NgoMIV site found at 7517
    Illegal NgoMIV site found at 9742
    Illegal NgoMIV site found at 10493
    Illegal AgeI site found at 4613
    Illegal AgeI site found at 5533
    Illegal AgeI site found at 6824
    Illegal AgeI site found at 9160
    Illegal AgeI site found at 9799
    Illegal AgeI site found at 10652
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1155


Design Notes

1.The sequence has a few restriction enzyme cutting sites, therefore checking is needed when we do enzyme digestion and ligation so that the sequence is correctly connected. 2.It is a very long sequence, below is few measures to take note during cloning.

  • The concentration agarose gel during gel electrophoresis should be about 0.8-1%
  • The enzyme digestion reaction time is advised to be elongated so that digestion could be more perfect.
  • The respective restriction enzyme at the both end of primer should not cut the sequence of another gene


Source

  • organism: Gluconacetobacter hansenii ATCC 23769
  • acs operon: GenBank AB091060.1
  • CMCax: GenBank EFG85213.1
  • Ccp: GenBank AAA16970.

References