Difference between revisions of "Part:BBa C0073:Design"

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===References===
 
===References===
 
* Identification of Functionally Important Residues in the DNA Binding Region of the MNT Repressor<br>
 
* Identification of Functionally Important Residues in the DNA Binding Region of the MNT Repressor<br>
−
Knight, KL; Sauer, RT<br>
+
**Knight, KL; Sauer, RT<br>
−
The Journal of Biological Chemistry, 264(23):13706-13710, 1989
+
**The Journal of Biological Chemistry, 264(23):13706-13710, 1989

Latest revision as of 22:04, 11 July 2006


mnt repressor (weak) from Salmonella phage P22 (+LVA)


Barcodes are discontinued, but one was appended to the sequence of this part. Composite parts using this part will include the barcode. More ...

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

dimeric repressor from enterobacteriophage p22 with dissociation constant of 3*10^-8.

sequence was translated with the E.Coli K12 codon usage table, and nucleotides 95-97 was changed from AAT to AAC to avoid the cut site GAATTC (94-99).

the mutation R10->K10 (ctg->aaa) decreases the binding affinity of the mnt protein to its operator by approximately 12,000 fold (Kd: 2.5e-12 to 3e-8)



Source

enterobacteriophage p22

References

  • Identification of Functionally Important Residues in the DNA Binding Region of the MNT Repressor
    • Knight, KL; Sauer, RT
    • The Journal of Biological Chemistry, 264(23):13706-13710, 1989