Coding
cyaA

Part:BBa_K845008

Designed by: Grégory Hansen   Group: iGEM12_Grenoble   (2012-09-26)
Revision as of 13:40, 30 September 2012 by Greghansen (Talk | contribs) (→‎Usage and Biology)

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cyaA with its natural RBS

cyaA gene (from BW25113 E. Coli) encodes the Adenyl cyclase protein which catalyses the conversion of ATP (Adenosine Tri-Phosphate) into cAMP (cyclic Adenosine Mono-Phosphate).

Our saferty note: https://static.igem.org/mediawiki/2012/4/48/CyaA.pdf


Usage and Biology

In Escherichia coli, cAMP is involved in carbon catabolite repression [1] and binds to the cAMP receptor protein (CRP). The corresponding complex (CRP-cAMP) is a transcriptional factor controlling the expression of more than 220 operons [2]. It has been known for a long time that E. coli actively exports cAMP into the growth medium [3][4].

The transcription of the cyaA gene and the crp gene is negatively regulated by CRP-cAMP [5] [6]. The translation of adenylate cyclase mRNA is ineffective as to prevent excessive synthesis of adenylate cyclase. This can be attributed to the fact that overproduction of cAMP is lethal to Escherichia coli possibly due to an accumulation of methylglyoxal [7] [8]. As we want to use paraBAD like an AND Gate and because high a concentration of cAMP is lethal for E. Coli we make our device in a BW25113 ΔcyaA.

http://www.uniprot.org/uniprot/P00936

http://ecocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10170

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 288
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 922
    Illegal SapI.rc site found at 412
    Illegal SapI.rc site found at 652


[edit]
Categories
//cds/enzyme
Parameters
None