DNA

Part:BBa_K1896020

Designed by: Bob Van Hove, Maarten Van Brempt   Group: iGEM16_UGent_Belgium   (2016-10-13)


pXW-GG

This part can be used to construct cloning vectors that are compatible with Golden Gate assembly.[1] Two diverging BsaI cloning sites are situated inbetween a weak to medium constitutive promoter+RBS and double terminator. One or more linear fragments can be cloned into this vector in an efficient one-pot restriction and ligation reaction. The outer sticky ends are GATG and TACT and the resulting constructs are indistinguishable from those created by standard Registry assembly methods, since the same scars are generated.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 77
    Illegal BsaI.rc site found at 66


References

  1. Engler, C., Gruetzner, R., Kandzia, R., & Marillonnet, S. (2009). Golden gate shuffling: a one-pot DNA shuffling method based on type IIs restriction enzymes. PloS one, 4(5), e5553.
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