DNA
RE

Part:BBa_J3101:Design

Designed by: Erin Zwack, Sabriya Rosemond   Group: iGEM06_Davidson   (2006-06-02)
Revision as of 09:31, 30 October 2008 by Rshetty (Talk | contribs) (→‎References)

Recombinational Enhancer (RE) for Hin/Hix inverting


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The C > T point mutation occurred during cloning. It is located in the spacer region between the Fis binding sites; therefore, we believe that the mutation will not impact RE function. There is an insertion immediately before the BioBrick suffix and several bases after the distal fis binding site that should affect RE function. RE is cloned in plasmid pSB1A2.

The BioBrick prefix and suffix on this part are not wildtype but the restriction sites are still viable.

Standard BioBrick Cloning Sites (Knight) 5'--GAATTC GCGGCCGC T TCTAGA G ----insert---- T ACTAGT A GCGGCCG CTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT C -------------- A TGATCA T CGCCGGC GACGTC--
BBa_J31001 Cloning Sites 5'--GAATTC GCGGCCGC T TCTAGA * ------RE------ G ACTAGT T GCGGCCGCCTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT * -------------- C TGATCA A CGCCGGCGGACGTC--

Prefix
There is no G spacer (*) between the XbaI and the RE.
Suffix
The T spacer between the RE and the SpeI site has changed to a G.
The A spacer between the SpeI and the NotI has changed to a T.
There is an extra C between the NotI site and the PstI site


Source

The Recombination Enhancer sequence from Salmonella typhimurium.

References

<biblio>

  1. Johnson pmid=2548848
  2. Haykinson pmd=8508775

</biblio>

  • Haykinson and Johnson (1993) DNA looping and the helical repeat in vitro and in vivo: effect of HU protein and enhancer location on Hin invertasome assembly

[http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=8508775]