Difference between revisions of "Part:BBa K1692021"

Revision as of 03:21, 20 September 2015

PanK + hybrid promoter phaCAB

S. aureus panK sequence added to hybrid promoter phaCAB operon

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 8
Illegal NheI site found at 31
Illegal NheI site found at 912
Illegal NheI site found at 935
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1884
Illegal BglII site found at 2709
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1190
Illegal NgoMIV site found at 1261
Illegal NgoMIV site found at 1861
Illegal NgoMIV site found at 2173
Illegal NgoMIV site found at 2452
Illegal NgoMIV site found at 3104
Illegal NgoMIV site found at 3126
Illegal AgeI site found at 418
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 4970

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1692021 short</partinfo>
@@ Line 7: / Line 6: @@
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Coenzyme A biosynthesis is a five-step process that is primarily regulated by the first enzyme in the pathway, pantothenate kinase (encoded by the gene PanK, which is also called CoaA) [4]. Pantothenate kinase stringently controls amount of coenzyme A produced in E. coli because the pantothenate kinase produced by E. coli is significantly inhibited by coenzyme A and its thioesters (such as acetyl-coA). Therefore, since we wanted to increase coenzyme A synthesis, we had to find a way to get around this negative feedback inhibition. Fortunately, the pantothenate kinase made in Staphylococcus aureus does not experience feedback inhibition from coenzyme A or its thioesters [5]. Indeed, this allows S. aureus to accumulate high levels of coenzyme A. We ordered the S. aureus pantothenate kinase sequence from IDT and inserted into the backbone PSB1C3 in front of the phaCAB operon designed by the Tokyo Tech iGEM team in 2012 with the hybrid promoter designed by the Imperial College iGEM team in 2013. The results section below shows that inserting the gene for pantothenate kinase allows for higher levels of P(3HB) production in E. coli.
 <!-- -->