Difference between revisions of "Part:BBa K3600002"

Latest revision as of 03:51, 28 October 2020

YCF1 promoter part plasmid

YCF1 promoter part plasmid was made in order to create a part plasmid which contains the YCF1 promoter and is compatible with the Yeast Toolkit. To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.

Usage and Biology

The YCF1 gene encodes the Yeast cadmium factor 1 ¹ and is required for yeast cells to better resist exposure to cadmium ². The YCF1 promoter is regulated by the transcription factor Yap1p. The binding site for Yap1p, in the promoter YCF1, is TTACTAA ⁴. Using the yeast toolkit for modular assembly, the YCF1 promoter plasmid was used as a part of type II .

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 935
Illegal EcoRI site found at 1665
Illegal XbaI site found at 944
Illegal XbaI site found at 1842
Illegal SpeI site found at 2810
Illegal PstI site found at 2819
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 935
Illegal EcoRI site found at 1665
Illegal SpeI site found at 2810
Illegal PstI site found at 2819
Illegal NotI site found at 923
Illegal NotI site found at 2829
21
INCOMPATIBLE WITH RFC[21]
Illegal suffix found in sequence at 2412
Illegal EcoRI site found at 935
Illegal EcoRI site found at 1665
Illegal BglII site found at 1280
Illegal BglII site found at 2283
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 935
Illegal EcoRI site found at 1665
Illegal XbaI site found at 944
Illegal XbaI site found at 1842
Illegal SpeI site found at 2810
Illegal PstI site found at 2819
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 935
Illegal EcoRI site found at 1665
Illegal XbaI site found at 944
Illegal XbaI site found at 1842
Illegal SpeI site found at 2810
Illegal PstI site found at 2819
Illegal NgoMIV site found at 2767
1000
COMPATIBLE WITH RFC[1000]

Measurements

References

[1] Bhuiyan, M. S. U. et al. Overexpression of a yeast cadmium factor 1 (YCF1) enhances heavy metal tolerance and accumulation in Brassica juncea. Plant Cell Tiss Organ Cult 105, 85–91 (2011).

[2] Li, Z.-S., Szczypka, M., Lu, Y.-P., Thiele, D. J. & Rea, P. A. The Yeast Cadmium Factor Protein (YCF1) Is a Vacuolar Glutathione S-Conjugate Pump. J. Biol. Chem. 271, 6509–6517 (1996).

[3] Lee, M. E., DeLoache, W. C., Cervantes, B. & Dueber, J. E. A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth. Biol. 4, 975–986 (2015).

[4] Kuge, S. & Jones, N. YAP1 dependent activation of TRX2 is essential for the response of Saccharomyces cerevisiae to oxidative stress by hydroperoxides. The EMBO Journal 13, 655–664 (1994).

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3600002 short</partinfo>
-.
+YCF1 promoter part plasmid was made in order to create a part plasmid which contains the YCF1 promoter and is compatible with the Yeast Toolkit. To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.
 ===Usage and Biology===
-* activator: Yap1p
+The YCF1 gene encodes the Yeast cadmium factor 1 <html><a href="#Bhuiyan2011"><sup>1</sup></a></html> and is required for yeast cells to better resist exposure to cadmium <html><a href="#Li1996"><sup>2</sup></a></html>. The YCF1 promoter is regulated by the transcription factor Yap1p. The binding site for Yap1p, in the promoter YCF1, is TTACTAA <html><a href="#Kuge1994"><sup>4</sup></a></html>. Using the yeast toolkit for modular assembly, the YCF1 promoter plasmid was used as a part of type II .
-*TXR2 gene encodes thioredoxin protein in Saccharomyces cerevisiae.<html><a href="#Kuge1994"><sup>1</sup></a></html>
-*The promoter TXR2 has been chosen in order to express fluorescent protein [ref] when the S.cervisiae is exposed to an oxidative stress.
-<!-- -->*The binding site for Yap1p, in the promoter YCF1, is TTACTAA.<html><a href="#Kuge1994"><sup>1</sup></a><!-- --></html>
+<!---->
@@ Line 16: / Line 17: @@
-<!-- Uncomment this to enable Functional Parameter display
+<!--
 ===Functional Parameters===
 <partinfo>BBa_K3600002 parameters</partinfo>
 <!-- -->
+===Measurements===
+===References===
+<HTML><a name=Bhuiyan2011 style="color:black">
+[1] Bhuiyan, M. S. U. et al. Overexpression of a yeast cadmium factor 1 (YCF1) enhances heavy metal tolerance and accumulation in Brassica juncea. Plant Cell Tiss Organ Cult 105, 85–91 (2011).
+</a></html>
+<HTML><a name=Li1996 style="color:black">
+[2] Li, Z.-S., Szczypka, M., Lu, Y.-P., Thiele, D. J. & Rea, P. A. The Yeast Cadmium Factor Protein (YCF1) Is a Vacuolar Glutathione S-Conjugate Pump. J. Biol. Chem. 271, 6509–6517 (1996).
+</a></html>
+<HTML><a name=Lee2015 style="color:black">
+[3] Lee, M. E., DeLoache, W. C., Cervantes, B. & Dueber, J. E. A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth. Biol. 4, 975–986 (2015).
+</a></html>
+<HTML><a name=Kuge1994 style="color:black">
+[4] Kuge, S. & Jones, N. YAP1 dependent activation of TRX2 is essential for the response of Saccharomyces cerevisiae to oxidative stress by hydroperoxides. The EMBO Journal 13, 655–664 (1994).
+</a></html>