Device

Part:BBa_M36532:Design

Designed by: Inderpreet Kaur Hayer   Group: Stanford BIOE44 - S11   (2015-10-24)
Revision as of 07:10, 8 December 2015 by Htale1 (Talk | contribs) (→‎Design Notes)

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 size limitations, PHO84, ACR 1, and ACR 2, this actuator was created in two 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 of both designs. This image shows the different components of the Arsenate Bioremediation Actuator.


Example implementation:

http://imageshack.com/a/img907/5935/3yeRre.png


http://imageshack.com/a/img907/254/OL6vcf.png

Source

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

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

In order to have a multi-protein expression cassette, we incorporated an Internal Ribosome Entry Site (IRES), which was a T2A sequence. We got the amino acid sequence for this part from the following site [http://blog.addgene.org/plasmids-101-multicistronic-vectors]

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 [5]

ACR 1: BBa_M36536 [6]

ACR 2: BBa_M36537 [7]

References

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