Difference between revisions of "Part:BBa M36532:Design"

(→‎Design Notes)
(→‎Design Notes)
Line 13: Line 13:
  
 
http://imageshack.com/a/img907/2517/DITZFh.png
 
http://imageshack.com/a/img907/2517/DITZFh.png
 +
 +
 +
[[File:plasmid2.jpg]]
  
 
http://imageshack.com/a/img907/9338/NX12gL.png
 
http://imageshack.com/a/img907/9338/NX12gL.png

Revision as of 03:00, 25 October 2015

Arsenate Bioremediation Actuator in S. Cerevisiae


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2711
    Illegal XbaI site found at 1680
    Illegal PstI site found at 1035
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2711
    Illegal PstI site found at 1035
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2711
    Illegal BglII site found at 1471
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2711
    Illegal XbaI site found at 1680
    Illegal PstI site found at 1035
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2711
    Illegal XbaI site found at 1680
    Illegal PstI site found at 1035
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2184


Design Notes

Below is an example of a possible implementation of BBa_M36532, the Arsenate Bioremediation Actuator. Due to the necessity of including genes PHO84, ACR 1, and ACR 2 and size limitations, it was necessary to design two separate constructs: one with the PHO84 (M36533)[1] and another with ACR 1 and 2 (M36534) [2]. In this example, a GAL1 inducible sensor has been attached upstream for both designs. This image shows the different components of the Arsenate Bioremediation Actuator. Part numbers for the Kozak sequence and the PHO84, ACR 1, and ACR 2 genes can be found in the following section.


Example implementation:

http://imageshack.com/a/img907/2517/DITZFh.png


Plasmid2.jpg

http://imageshack.com/a/img907/9338/NX12gL.png

Source

DNA 2.0 Plasmid (pD1201) with GAL1 inducible promoter, high copy number, and Uracil as the selectable marker. [3]

DNA 2.0 Plasmid (pD1204) with GAL1 inducible promoter, high copy number, and Uracil as the selectable marker. [4]

RBS consensus yeast Kozak sequence taken from the Silver Lab, BBa_J63003[5] , was used for RBS1 and 2 on both constructs.

Naturally occurring genes in S. cerevisiae: Pho84, ACR 1 and 2 Sequences were referenced from the yeast genome website and then were entered into the registry.

PHO84: BBa_M36535 [6]

ACR 1: BBa_M36536 [7]

ACR 2: BBa_M36537 [8]

References

Engineering of Saccharomyces cerevisiae for Bioremediation of Arsenate and Development of Yeast Vector Tools [http://gradworks.umi.com/35/02/3502760.htm]