Part:BBa_E0033:Design

LacZ alpha fragment; complements matching N-terminal deletion mutant (lacZ-omega)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 93
  Illegal XhoI site found at 144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Restriction sites (modified)
76-79 (gcc to gca) 79-81 (gct to gca) 85-87 (gaa to gag) 106-108 (tgc to tgt) 109-111(agg to aga)

• RS 11:45, 20 July 2007 (EDT): As currently specified, this sequence appears to be wrong. There should be a T before the AATAA (last 5 nucleotides) in order for the double stop codon to be in frame with the rest of the protein. I am not sure if this error is only in the sequence entered into the Registry or if the physical DNA is also missing a T as well.
  • RS 18:00, 2 August 2007 (EDT): Sequencing of BBa_E0433 indicates that the sequence in the Registry does reflect the physical part sequence. So this parts lacks an in frame stop codon. This may render lacZalpha nonfunctional.
• RS 12:55, 20 July 2007 (EDT): This appears to be derived from a [http://www.addgene.org/pgvec1?f=c&cmd=showvecinfo&vectorid=5652 pBluescript II (SK-)] or similar plasmid. It contains an insert in the lacZ N terminal fragment to enable construction of a multiple cloning site within the lacZalpha coding sequence. The lacZ coding sequence is frequently used as a screening mechanism when cloning DNA fragments into vectors (blue-white selection). It appears that the E0033 part designers simply took the lacZalpha fragment from a cloning vector and mutated out the relevant restriction enzyme sites.

Source

www.ncbi.nlm.nih.gov

References

The lacZ-alpha/omega complementation system is well-described [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture4/Lecture4.html here].