U.S. patent application number 12/967320 was filed with the patent office on 2011-06-09 for reporter assay screens for protein targets in saccharomyces cerevisiae.
Invention is credited to Marvin Bayne, Thomas J. Hosted, Scott Walker.
Application Number | 20110136128 12/967320 |
Document ID | / |
Family ID | 36206622 |
Filed Date | 2011-06-09 |
United States Patent
Application |
20110136128 |
Kind Code |
A1 |
Hosted; Thomas J. ; et
al. |
June 9, 2011 |
REPORTER ASSAY SCREENS FOR PROTEIN TARGETS IN SACCHAROMYCES
CEREVISIAE
Abstract
The present invention comprises responsive promoters along with
screening methods which make use of the promoters in order to
identify anti-fungal substances.
Inventors: |
Hosted; Thomas J.; (Summit,
NJ) ; Walker; Scott; (Basking Ridge, NJ) ;
Bayne; Marvin; (Westfield, NJ) |
Family ID: |
36206622 |
Appl. No.: |
12/967320 |
Filed: |
December 14, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11226605 |
Sep 14, 2005 |
7872115 |
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12967320 |
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60609940 |
Sep 15, 2004 |
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Current U.S.
Class: |
435/6.13 ;
435/254.11; 435/320.1; 536/24.1 |
Current CPC
Class: |
C12Q 1/18 20130101; C12Q
2600/158 20130101; C12Q 1/6897 20130101; C07K 14/395 20130101 |
Class at
Publication: |
435/6.13 ;
536/24.1; 435/320.1; 435/254.11 |
International
Class: |
C12Q 1/68 20060101
C12Q001/68; C07H 21/04 20060101 C07H021/04; C12N 15/63 20060101
C12N015/63; C12N 1/15 20060101 C12N001/15 |
Claims
1. An isolated hybrid polynucleotide comprising a member selected
from the group consisting of: S. cerevisiae YOR387C Promoter, S.
cerevisiae YMR175W Promoter, S. cerevisiae YFR026C Promoter, S.
cerevisiae YJL153C Promoter, S. cerevisiae YPL033C Promoter, S.
cerevisiae YOL058W Promoter, S. cerevisiae YOR255W Promoter, S.
cerevisiae YDR250C Promoter, S. cerevisiae YDR446W Promoter, S.
cerevisiae YDR536W Promoter, S. cerevisiae YOR255W Promoter, S.
cerevisiae YDL243C Promoter S. cerevisiae YDR256C Promoter, S.
cerevisiae YFL020C Promoter, S. cerevisiae YPL205C Promoter, S.
cerevisiae YGL205W Promoter, S. cerevisiae YGL117W Promoter, S.
cerevisiae YHR029C Promoter, S. cerevisiae YML116W Promoter S.
cerevisiae YJR109C Promoter, S. cerevisiae YCL030C Promoter, S.
cerevisiae YLR338W Promoter, S. cerevisiae YIL066W-A Promoter, S.
cerevisiae YNL093W Promoter, S. cerevisiae YLR381W Promoter, S.
cerevisiae YKL159C Promoter, S. cerevisiae YIL058W Promoter, S.
cerevisiae YKR037C Promoter, S. cerevisiae YNL279W Promoter, S.
cerevisiae YOR032C Promoter, S. cerevisiae YML058W Promoter, S.
cerevisiae YMR303C Promoter, S. cerevisiae YJL153C Promoter S.
cerevisiae YLR092W Promoter, S. cerevisiae YFL052W Promoter, S.
cerevisiae YIR017C Promoter, S. cerevisiae YLL062C Promoter, S.
cerevisiae YMR323W Promoter S. cerevisiae YPL171C Promoter, S.
cerevisiae YER065C Promoter, S. cerevisiae YDR114C Promoter, S.
cerevisiae YHR082C Promoter, S. cerevisiae YDL126C Promoter S.
cerevisiae YBR106W Promoter, S. cerevisiae YLL043W Promoter, S.
cerevisiae YPR108W Promoter, S. cerevisiae YIL041W Promoter, S.
cerevisiae YJL166W Promoter S. cerevisiae YDR165W Promoter, S.
cerevisiae YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S.
cerevisiae YHR012W Promoter, S. cerevisiae YKL053W Promoter, S.
cerevisiae YKL196C Promoter, S. cerevisiae YDL103C Promoter S.
cerevisiae YLR427W Promoter, S. cerevisiae YLL020C Promoter and S.
cerevisiae YBL065W Promoter; operably linked to a reporter.
2. An isolated polynucleotide of claim 1 wherein said promoter
comprises a nucleotide sequence selected from the group consisting
of SEQ ID NOs: 1-41 and 52-68; and/or the reporter comprises a
nucleotide sequence selected from the group consisting of SEQ ID
NOs: 42-51.
3. An isolated vector comprising a polynucleotide of claim 1.
4. An isolated host cell comprising a vector of claim 3.
5. An isolated plasmid polynucleotide comprising a nucleotide
sequence selected from the group consisting of SEQ ID NOs:
85-88.
6. A method for identifying a substance that inhibits fungal cell
growth comprising: (a) introducing a hybrid comprising a promoter
selected from the group consisting of S. cerevisiae YOR387C
Promoter, S. cerevisiae YMR175W Promoter, S. cerevisiae YFR026C
Promoter, S. cerevisiae YJL153C Promoter, S. cerevisiae YPL033C
Promoter, S. cerevisiae YOL058W Promoter, S. cerevisiae YOR255W
Promoter, S. cerevisiae YDR250C Promoter, S. cerevisiae YDR446W
Promoter, S. cerevisiae YDR536W Promoter, S. cerevisiae YOR255W
Promoter, S. cerevisiae YDL243C Promoter S. cerevisiae YDR256C
Promoter, S. cerevisiae YFL020C Promoter, S. cerevisiae YPL205C
Promoter, S. cerevisiae YGL205W Promoter, S. cerevisiae YGL117W
Promoter, S. cerevisiae YHR029C Promoter, S. cerevisiae YML116W
Promoter S. cerevisiae YJR109C Promoter, S. cerevisiae YCL030C
Promoter, S. cerevisiae YLR338W Promoter, S. cerevisiae YIL066W-A
Promoter, S. cerevisiae YNL093W Promoter, S. cerevisiae YLR381W
Promoter, S. cerevisiae YKL159C Promoter, S. cerevisiae YIL058W
Promoter, S. cerevisiae YKR037C Promoter, S. cerevisiae YNL279W
Promoter, S. cerevisiae YOR032C Promoter, S. cerevisiae YML058W
Promoter, S. cerevisiae YMR303C Promoter, S. cerevisiae YJL153C
Promoter S. cerevisiae YLR092W Promoter, S. cerevisiae YFL052W
Promoter, S. cerevisiae YIR017C Promoter, S. cerevisiae YLL062C
Promoter, S. cerevisiae YMR323W Promoter, S. cerevisiae YPL171C
Promoter, S. cerevisiae YER065C Promoter and S. cerevisiae YDR114C
Promoter; operably linked to a reporter; into a suitable fungal
cell; (b) contacting the cell with a substance to be tested for the
ability to inhibit fungal cell growth; and (c) detecting expression
driven by the promoter in the hybrid; wherein the substance is
identified as a fungal growth inhibitor if the substance modulates
expression from the promoter.
7. A method of claim 6 wherein the promoter comprises a nucleotide
sequence selected from the group consisting of SEQ ID NOs: 1-41 and
68.
8. A method of claim 6 further comprising a negative-control
comprising: (i) introducing the hybrid into a suitable fungal cell;
(ii) contacting the cell with a blank substance which is known to
not modulate expression from the promoter in the hybrid; and (iii)
detecting expression driven by the promoter in the hybrid; and (iv)
comparing the expression level from the promoter in the cell
contacted with the blank substance with the expression level from
the promoter in the cell contacted with the substance to be tested
for the ability to inhibit fungal cell growth.
9. A method of claim 6 further comprising a positive-control
comprising: (i) introducing the hybrid into a suitable fungal cell;
(ii) contacting the cell with a positive-control which is known to
modulate expression from the promoter in the hybrid; (iii)
detecting expression driven by the promoter in the hybrid; and (iv)
comparing the expression level from the promoter in the cell
contacted with the positive-control substance with the expression
level from the promoter in the cell contacted with the substance to
be tested for the ability to inhibit fungal cell growth.
10. A method for identifying a substance that inhibits fungal cell
growth comprising: (a) introducing, into a suitable fungal cell, a
first hybrid comprising a first promoter selected from the group
consisting of: S. cerevisiae YOR387C Promoter, S. cerevisiae
YMR175W Promoter, S. cerevisiae YFR026C Promoter, S. cerevisiae
YJL153C Promoter, S. cerevisiae YPL033C Promoter, S. cerevisiae
YOL058W Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae
YDR250C Promoter, S. cerevisiae YDR446W Promoter, S. cerevisiae
YDR536W Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae
YDL243C Promoter S. cerevisiae YDR256C Promoter, S. cerevisiae
YFL020C Promoter, S. cerevisiae YPL205C Promoter, S. cerevisiae
YGL205W Promoter, S. cerevisiae YGL117W Promoter, S. cerevisiae
YHR029C Promoter, S. cerevisiae YML116W Promoter S. cerevisiae
YJR109C Promoter, S. cerevisiae YCL030C Promoter, S. cerevisiae
YLR338W Promoter, S. cerevisiae YIL066W-A Promoter, S. cerevisiae
YNL093W Promoter, S. cerevisiae YLR381W Promoter, S. cerevisiae
YKL159C Promoter, S. cerevisiae YIL058W Promoter, S. cerevisiae
YKR037C Promoter, S. cerevisiae YNL279W Promoter, S. cerevisiae
YOR032C Promoter, S. cerevisiae YML058W Promoter, S. cerevisiae
YMR303C Promoter, S. cerevisiae YJL153C Promoter S. cerevisiae
YLR092W Promoter, S. cerevisiae YFL052W Promoter, S. cerevisiae
YIR017C Promoter, S. cerevisiae YLL062C Promoter, S. cerevisiae
YMR323W Promoter S. cerevisiae YPL171C Promoter, S. cerevisiae
YER065C Promoter, S. cerevisiae YDR114C Promoter and S. cerevisiae
YBL065W Promoter; operably linked to a first reporter; and a second
hybrid comprising a second promoter selected from the group
consisting of: S. cerevisiae YHR082C Promoter, S. cerevisiae
YDL126C Promoter S. cerevisiae YBR106W Promoter, S. cerevisiae
YLL043W Promoter, S. cerevisiae YPR108W Promoter, S. cerevisiae
YIL041W Promoter, S. cerevisiae YJL166W Promoter S. cerevisiae
YDR165W Promoter, S. cerevisiae YLR330 W Promoter, S. cerevisiae
YDL058W Promoter, S. cerevisiae YHR012W Promoter, S. cerevisiae
YKL053W Promoter, S. cerevisiae YKL196C Promoter, S. cerevisiae
YDL103C Promoter S. cerevisiae YLR427W Promoter and S. cerevisiae
YLL020C Promoter; operably linked to a second reporter; wherein
said first and second reporters are different; (b) contacting the
cell with a substance to be tested for the ability to inhibit
fungal cell growth; and (c) detecting expression driven from the
first and second promoters; wherein the substance is identified as
a fungal growth inhibitor if the substance causes the first
promoter expression level to increase or decrease in relation to
the expression level from the second reporter.
11. The method of claim 10 further comprising a negative-control
comprising: (i) introducing said first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a blank
substance which is known to not modulate expression from the
promoters in the hybrids; (iii) detecting expression driven by the
promoters in the hybrids; and (iv) comparing the expression level
from the first promoter in relation to the expression level from
the second promoter in the cell contacted with the blank substance
with the expression level from the first promoter in relation to
the expression level from the second promoter in the cell contacted
with the substance to be tested for the ability to inhibit fungal
cell growth.
12. The method of claim 10 further comprising a positive-control
comprising: (i) introducing said first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a
positive-control substance which is known to modulate expression
from the promoters in the hybrids; (iii) detecting expression
driven by the promoters in the hybrids; and (iv) comparing the
expression level from the first promoter in relation to the
expression level from the second promoter in the cell contacted
with the positive-control substance with the expression level from
the first promoter in relation to the expression level from the
second promoter in the cell contacted with the substance to be
tested for the ability to inhibit fungal cell growth.
13. A method of claim 10 wherein the first promoter comprises a
nucleotide sequence selected from the group consisting of SEQ ID
NOs: 1-41 and 68; the second promoter comprises a nucleotide
sequence selected from the group consisting of SEQ ID NOs: 52-67;
and/or the reporter comprises a nucleotide sequence selected from
the group consisting of SEQ ID NOs: 42-51.
14. A method for determining whether a substance inhibits growth of
a fungal cell comprising: (a) introducing a hybrid comprising a
promoter selected from the group consisting of: S. cerevisiae
YHR082C Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae
YBR106W Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae
YPR108W Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae
YJL166W Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae
YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae
YHR012W Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae
YKL196C Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae
YLR427W Promoter and S. cerevisiae YLL020C Promoter; operably
linked to a reporter into a suitable fungal cell; (b) contacting
the cell with a substance to be tested for the ability to inhibit
growth of the cell; and (c) detecting signal from the reporter;
wherein the substance is identified as a fungal growth inhibitor if
the substance causes a reduction in the signal from the
reporter.
15. The method of claim 14 further comprising a negative-control
comprising: (i) introducing the hybrid into a suitable fungal cell;
(ii) contacting the cell with a blank substance which is known to
not inhibit fungal cell growth; (iii) detecting signal from the
reporter in the hybrid; and (iv) comparing the signal from the
reporter in the cell contacted with the blank substance with the
signal from the reporter in the cell contacted with the substance
to be tested for the ability to inhibit fungal cell growth.
16. The method of claim 14 further comprising a positive-control
comprising: (i) introducing the hybrid into a suitable fungal cell;
(ii) contacting the cell with a positive-control substance which is
known to inhibit fungal cell growth; (iii) detecting signal from
the reporter in the hybrid; and (iv) comparing the signal from the
reporter in the cell contacted with the positive-control substance
with the signal from the reporter in the cell contacted with the
substance to be tested for the ability to inhibit fungal cell
growth.
17. A method of claim 14 wherein the promoter comprises a
nucleotide sequence selected from the group consisting of SEQ ID
NOs: 52-67; and/or the reporter comprises a nucleotide sequence
selected from the group consisting of SEQ ID NOs: 42-51.
18. A method for determining whether a substance inhibits growth of
a fungal cell comprising: (a) introducing a first hybrid comprising
a first promoter selected from the group consisting of: S.
cerevisiae YHR082C Promoter, S. cerevisiae YDL126C Promoter S.
cerevisiae YBR106W Promoter, S. cerevisiae YLL043W Promoter, S.
cerevisiae YPR108W Promoter, S. cerevisiae YIL041W Promoter, S.
cerevisiae YJL166W Promoter S. cerevisiae YDR165W Promoter, S.
cerevisiae YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S.
cerevisiae YHR012W Promoter, S. cerevisiae YKL053W Promoter, S.
cerevisiae YKL196C Promoter, S. cerevisiae YDL103C Promoter S.
cerevisiae YLR427W Promoter and S. cerevisiae YLL020C Promoter;
operably linked to a first reporter; and a second hybrid comprising
a second promoter selected from the group consisting of: S.
cerevisiae YHR082C Promoter, S. cerevisiae YDL126C Promoter S.
cerevisiae YBR106W Promoter, S. cerevisiae YLL043W Promoter, S.
cerevisiae YPR108W Promoter, S. cerevisiae YIL041W Promoter, S.
cerevisiae YJL166W Promoter S. cerevisiae YDR165W Promoter, S.
cerevisiae YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S.
cerevisiae YHR012W Promoter, S. cerevisiae YKL053W Promoter, S.
cerevisiae YKL196C Promoter, S. cerevisiae YDL103C Promoter S.
cerevisiae YLR427W Promoter and S. cerevisiae YLL020C Promoter;
operably linked to a second reporter into a suitable fungal cell;
wherein said first and second promoter are different and wherein
said first and second reporter are different; (b) contacting the
cell with a substance to be tested for the ability to inhibit
growth of the cell; and (c) detecting signal from the reporters;
wherein the substance is identified as a fungal growth inhibitor if
the substance causes a reduction in the signal from both the first
and second reporters.
19. The method of claim 18 further comprising a negative-control
comprising: (i) introducing the first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a blank
substance which is known to not inhibit fungal cell growth; (iii)
detecting signal from first and second reporters in the hybrids;
and (iv) comparing the signal from the first and second reporters
in the cell contacted with the blank substance with the signal from
the first and second reporters in the cell contacted with the
substance to be tested for the ability to inhibit fungal cell
growth.
20. The method of claim 18 further comprising a positive-control
comprising: (i) introducing the first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a positive
control substance which is known to inhibit fungal cell growth;
(iii) detecting signal from first and second reporters in the
hybrids; and (iv) comparing the signal from the first and second
reporters in the cell contacted with the positive control substance
with the signal from the first and second reporters in the cell
contacted with the substance to be tested for the ability to
inhibit fungal cell growth.
21. A method of claim 18 wherein said promoters comprise a
nucleotide sequence selected from the group consisting of SEQ ID
NOs: 52-67; and/or the reporter comprises a nucleotide sequence
selected from the group consisting of SEQ ID NOs: 42-51.
22. A method for determining whether a substance inhibits growth of
a malignant cell comprising (a) introducing, into a suitable fungal
cell, a first hybrid comprising a first promoter selected from the
group consisting of: S. cerevisiae YLR338W, S. cerevisiae YIL066W,
S. cerevisiae YNL093W, S. cerevisiae YDR250C, S. cerevisiae
YLR381W, S. cerevisiae YBL065W, S. cerevisiae YMR323W and S.
cerevisiae YPL171C; operably linked to a first reporter; and a
second hybrid comprising a second promoter selected from the group
consisting of: S. cerevisiae YHR082C Promoter, S. cerevisiae
YDL126C Promoter S. cerevisiae YBR106W Promoter, S. cerevisiae
YLL043W Promoter, S. cerevisiae YPR108W Promoter, S. cerevisiae
YIL041W Promoter, S. cerevisiae YJL166W Promoter S. cerevisiae
YDR165W Promoter, S. cerevisiae YLR330 W Promoter, S. cerevisiae
YDL058W Promoter, S. cerevisiae YHR012W Promoter, S. cerevisiae
YKL053W Promoter, S. cerevisiae YKL196C Promoter, S. cerevisiae
YDL103C Promoter S. cerevisiae YLR427W Promoter and S. cerevisiae
YLL020C Promoter; operably linked to a second reporter; (b)
contacting the cell with a substance to be tested for the ability
to inhibit growth of a malignant cell; and (c) detecting expression
driven by the first and second promoters; wherein the substance is
identified as a malignant cell growth inhibitor if the substance
causes the expression level from the first promoter to increase or
decrease in relation to the expression level from the second
promoter.
23. The method of claim 22 wherein the first promoter comprises a
nucleotide sequence selected from the group consisting of SEQ ID
NOs: 8, 22-25, 38, 39 and 68; and/or the second promoter comprises
a nucleotide sequence selected from the group consisting of SEQ ID
NOs: 52-67.
24. The method of claim 22 further comprising a negative-control
comprising: (i) introducing said first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a blank
substance which is known to not modulate expression from the
promoters in the hybrids; and (iii) detecting expression driven by
the promoters in the hybrids; (iv) comparing the expression level
from the first promoter in realtion to the expression level from
the second promoter in the cell contacted with the blank substance
with the expression level from the first promoter in realtion to
the expression level from the second promoter in the cell contacted
with the substance to be tested for the ability to inhibit fungal
cell growth.
25. The method of claim 22 further comprising a positive-control
comprising: (i) introducing said first and second hybrids into a
suitable fungal cell; (ii) contacting the cell with a
positive-control substance which is known to modulate expression
from the promoters in the hybrids; (iii) detecting expression
driven by the promoters in the hybrids; and (iv) comparing the
expression level from the first promoter in realtion to the
expression level from the second promoter in the cell contacted
with the positive control substance with the expression level from
the first promoter in realtion to the expression level from the
second promoter in the cell contacted with the substance to be
tested for the ability to inhibit fungal cell growth.
Description
[0001] The present application is a continuation of U.S. patent
application Ser. No. 11/226,605; filed Sep. 14, 2005 which claims
the benefit of U.S. provisional patent application No. 60/609,940;
filed Sep. 15, 2004; each of which is herein incorporated by
reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to compositions and methods
for screening for compounds which modulate the function of an
essential gene, for example, in fungal cells.
BACKGROUND OF THE INVENTION
[0003] High throughput assays, for identifying candidate compounds
which inhibit one or more target proteins in an organism, are most
efficient when they include an easily identifiable signal for
identifying compounds with the desired inhibitory activity. One
such well known system utilizes a responsive promoter, which is
activated by the inactivation of the target by a candidate
compound, and which is fused to an easily identifiable reporter.
When the target protein is inactivated by a candidate compound, the
reporter is expressed and the corresponding compound is identified
as inhibitory. Such responsive promoters can be identified simply
by identifying promoters which are activated by the genetic
activation or inactivation of a target gene of interest (e.g., by
construction of a temperature-sensitive mutation in the gene).
[0004] Responsive promoter screening assays have been used to
identify anti-fungal compounds such as azoles which inhibit the
egosterol biosynthesis pathway. U.S. Pat. No. 5,527,687; Dixon et
al., J. Steroid Biochem. Mol. Biol. 1997 June; 62(2-3):165-71. In
addition, screens have been developed to screen for inhibitors of
protein secretion, membrane stability, protein synthesis and cell
wall integrity. Bianchi et al., Appl Environ. Microbiol. 1999
November; 65(11):5023-7; Alksne et al., Antimicrob. Agents
Chemother. 2000 June; 44(6):1418-27; Shapiro et al., Antimicrob.
Agents Chemother. 2002 August; 46(8):2490-7; Sun et al., J.
Antibiot. (Tokyo). 2002; 55(3): 279-287. These screens are
convenient for high-throughput assay systems requiring short
incubation times, sub-lethal compound concentrations and the
selective induction of the reporter fusion indicates that the
compound is perturbing the pathway of interest. Fischer et al.,
Genome Res. 2004 January; 14(1):90-8.
[0005] Although some responsive promoters are known and they have
been used to identify anti-microbial substances, there still exists
a need in the art for the identification of new responsive
promoters which may be used to identify other substances of
clinical interest.
SUMMARY OF THE INVENTION
[0006] The present invention addresses the need in the art for
additional responsive promoters for use in assays to identify novel
anti-fungal and anti-cancer substances.
[0007] The present invention provides an isolated hybrid
polynucleotide comprising a member selected from the group
consisting of S. cerevisiae YOR387C Promoter, S. cerevisiae YMR175W
Promoter, S. cerevisiae YFR026C Promoter, S. cerevisiae YJL153C
Promoter, S. cerevisiae YPL033C Promoter, S. cerevisiae YOL058W
Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae YDR250C
Promoter, S. cerevisiae YDR446W Promoter, S. cerevisiae YDR536W
Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae YDL243C
Promoter S. cerevisiae YDR256C Promoter, S. cerevisiae YFL020C
Promoter, S. cerevisiae YPL205C Promoter, S. cerevisiae YGL205W
Promoter, S. cerevisiae YGL117W Promoter, S. cerevisiae YHR029C
Promoter, S. cerevisiae YML116W Promoter S. cerevisiae YJR109C
Promoter, S. cerevisiae YCL030C Promoter, S. cerevisiae YLR338W
Promoter, S. cerevisiae YIL066W-A Promoter, S. cerevisiae YNL093W
Promoter, S. cerevisiae YLR381W Promoter, S. cerevisiae YKL159C
Promoter, S. cerevisiae YIL058W Promoter, S. cerevisiae YKR037C
Promoter, S. cerevisiae YNL279W Promoter, S. cerevisiae YOR032C
Promoter, S. cerevisiae YML058W Promoter, S. cerevisiae YMR303C
Promoter, S. cerevisiae YJL153C Promoter S. cerevisiae YLR092W
Promoter, S. cerevisiae YFL052W Promoter, S. cerevisiae YIR017C
Promoter, S. cerevisiae YLL062C Promoter, S. cerevisiae YMR323W
Promoter S. cerevisiae YPL171C Promoter, S. cerevisiae YER065C
Promoter, S. cerevisiae YDR114C Promoter, S. cerevisiae YHR082C
Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae YBR106W
Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae YPR108W
Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae YJL166W
Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae YLR330 W
Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae YHR012W
Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae YKL196C
Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae YLR427W
Promoter, S. cerevisiae YLL020C Promoter and S. cerevisiae YBL065W
Promoter (e.g., SEQ ID NOs: 1-41, 52-68); optionally operably
linked to a reporter (e.g., S. cerevisiae ADE2, S. cerevisiae LYS2,
S. cerevisiae TRP1, S. cerevisiae LEU2, S. cerevisiae URA3, S.
cerevisiae HIS3, Aequorea victoria GFP mutant 3, Renilla
luciferase, Photinus pyralis luciferase, Photinus pyralis
luciferase slk mutant, Vibrio fischeri luxA, Vibrio fischeri luxB,
Vibrio fischeri luxC, Vibrio fischeri luxD, Vibrio fischeri luxE,
Vibrio fischeri luxAB, Vibrio fischeri luxCDABE, Vibrio harveyi
luxA, Vibrio harveyi luxB, Vibrio harveyi luxC, Vibrio harveyi
luxD, Vibrio harveyi luxE, Vibrio harveyi luxAB, Vibrio harveyi
luxCDABE, Photorhabdus luminscens LuxA, Photorhabdus luminscens
LuxB, Photorhabdus luminscens LuxC, Photorhabdus luminscens LuxD,
Photorhabdus luminscens LuxE, Photorhabdus luminscens LuxCDABE, E.
coli lacZ, the Aequorea victoria Aequorin gene, KanMX, pat1, nat1,
hph, CAT, Sh Ble, GUS, CYH2 or CAN1 (e.g., SEQ ID NOs: 42-51)).
Also within the scope of the invention is an embodiment comprising
one of the foregoing promoters, operably linked to a reporter with
the proviso that the reporter is not the open reading frame that is
naturally located downstream of the promoter in the S. cerevisiae
genome. An embodiment of the invention comprises a vector
comprising a promoter of the present invention or hybrid thereof
along with a host cell comprising the vector. In another embodiment
of the invention, the vector in the host cell is episomal or
integrated into a chromosome of the host cell.
[0008] The present invention also provides the isolated plasmids
pSPRT47; pSPRT50; pSPRT190 and pSPRT192 (SEQ ID NOs.:85-88).
[0009] The present invention also provides a method for identifying
a substance that inhibits fungal cell growth comprising (a)
introducing a hybrid comprising a promoter selected from the group
consisting of S. cerevisiae YOR387C Promoter, S. cerevisiae YMR175W
Promoter, S. cerevisiae YFR026C Promoter, S. cerevisiae YJL153C
Promoter, S. cerevisiae YPL033C Promoter, S. cerevisiae YOL058W
Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae YDR250C
Promoter, S. cerevisiae YDR446W Promoter, S. cerevisiae YDR536W
Promoter, S. cerevisiae YOR255W Promoter, S. cerevisiae YDL243C
Promoter S. cerevisiae YDR256C Promoter, S. cerevisiae YFL020C
Promoter, S. cerevisiae YPL205C Promoter, S. cerevisiae YGL205W
Promoter, S. cerevisiae YGL117W Promoter, S. cerevisiae YHR029C
Promoter, S. cerevisiae YML116W Promoter S. cerevisiae YJR109C
Promoter, S. cerevisiae YCL030C Promoter, S. cerevisiae YLR338W
Promoter, S. cerevisiae YIL066W-A Promoter, S. cerevisiae YNL093W
Promoter, S. cerevisiae YLR381W Promoter, S. cerevisiae YKL159C
Promoter, S. cerevisiae YIL058W Promoter, S. cerevisiae YKR037C
Promoter, S. cerevisiae YNL279W Promoter, S. cerevisiae YOR032C
Promoter, S. cerevisiae YML058W Promoter, S. cerevisiae YMR303C
Promoter, S. cerevisiae YJL153C Promoter S. cerevisiae YLR092W
Promoter, S. cerevisiae YFL052W Promoter, S. cerevisiae YIR017C
Promoter, S. cerevisiae YLL062C Promoter, S. cerevisiae YMR323W
Promoter S. cerevisiae YPL171C Promoter, S. cerevisiae YER065C
Promoter, S. cerevisiae YDR114C Promoter and S. cerevisiae YBL065W
Promoter (e.g., SEQ ID NOs: 1-41 or 68) operably linked to a
reporter (e.g., S. cerevisiae ADE2, S. cerevisiae LYS2, S.
cerevisiae TRP1, S. cerevisiae LEU2, S. cerevisiae URA3, S.
cerevisiae HIS3, Aequorea victoria GFP mutant 3, Renilla
luciferase, Photinus pyralis luciferase, Photinus pyralis
luciferase slk mutant, Vibrio fischeri luxA, Vibrio fischeri luxB,
Vibrio fischeri luxC, Vibrio fischeri luxD, Vibrio fischeri luxE,
Vibrio fischeri luxAB, Vibrio fischeri luxCDABE, Vibrio harveyi
luxA, Vibrio harveyi luxB, Vibrio harveyi luxC, Vibrio harveyi
luxD, Vibrio harveyi luxE, Vibrio harveyi luxAB, Vibrio harveyi
luxCDABE, Photorhabdus luminscens LuxA, Photorhabdus luminscens
LuxB, Photorhabdus luminscens LuxC, Photorhabdus luminscens LuxD,
Photorhabdus luminscens LuxE, Photorhabdus luminscens LuxCDABE, E.
coli lacZ, the Aequorea victoria Aequorin gene, KanMX, pat1, nat1,
hph, CAT, Sh Ble, GUS, CYH2 or CAN1 (e.g., SEQ ID NOs: 42-51)) into
a suitable fungal cell (e.g., Absidia corymbifera; Absidia spp;
Acremonium spp; Ajellomyces capsulatus; Ajellomyces dermatitidis;
Alternaria spp; Aphanoascus fulvescens; Apophysomyces spp;
Arthroderma benhamiae; Arthroderma fulvum; Arthroderma gypseum;
Arthroderma incurvatum; Arthroderma otae; Arthroderma
vanbreuseghemii; Aspergillus flavus; Aspergillus fumigatus;
Aspergillus glaucus; Aspergillus nidulans; Aspergillus niger;
Aspergillus oryzae; Aspergillus spp; Aspergillus sydowi;
Aspergillus terreus; Aspergillus ustus; Aspergillus versicolor;
Aureobasidium pullulans; Basidiomycetes; Beauveria spp; Bipolaris
hawaiiensis; Bipolaris spicifera; Bipolaris spp; Bjerkandera
adusta; Blastomyces dermatitidis; Blastoschizomyces capitatus;
Candida albicans; Candida beigelii; Candida colluculosa; Candida
dubliniensis; Candida dubliniensis; Candida famata; Candida famata;
Candida glabrata; Candida guilliermondii; Candida haemulonii;
Candida holmii; Candida inconspicua; Candida intermedia; Candida
keyfrr; Candida krusei; Candida krusei; Candida lambica; Candida
lipolytica; Candida lusitaniae; Candida maris; Candida melibiosica;
Candida norvegensis; Candida parapsilosis; Candida parapsilosis;
Candida pelliculosa; Candida pelliculosa; Candida pseudotropicalis;
Candida pulcherrima; Candida rugosa; Candida sake; Candida
sphaerica; Candida spp; Candida stellatoidea; Candida tropicalis;
Candida tropicalis; Candida viswanathii; Candida zeylanoides;
Chrysosporium spp; Cladophialophora bantiana; Cladophialophora
carrionii; Cladosporium spp; Coccidioides immitis; Cokeromyces
recurvatus; Coprinus spp; Cryptococcus albidus; Cryptococcus
gattii; Cryptococcus laurentii; Cryptococcus neoformans;
Cunninghamella bertholletiae; Cunninghamella spp; Curvularia
lunata; Curvularia spp; Dekkera bruxellensis; Epidermophyton
floccosum; Epidermophyton floccosum; Exophiala dermatitidis;
Exophiala jeanselmei; Exophiala moniliae; Exserohilum rostratum;
Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium dimerum;
Fusarium moniliforme; Fusarium oxysporum; Fusarium proliferatum;
Fusarium solani; Fusarium spp; Geotrichum candidum; Geotrichum spp;
Histoplasma capsulatum; Hortaea werneckii; Issatschenkia
orientalis; Kluveromyces lactis; Kluyveromyces marxianus; Madurella
grisae; Malassezia furfur; Malassezia globosa; Malassezia obtusa;
Malassezia pachydermatis; Malassezia restricta; Malassezia
slooffiae; Malassezia sympodialis; Metarrhizium anisopliae;
Microsporum audouinii; Microsporum canis; Microsporum fulvum;
Microsporum gypseum; Microsporum persicolor; Mucor circinelloides;
Mucor hiemalis; Mucor racemosus; Mucor rouxii; Mucor spp;
Nattrassia mangiferae; Nectria haematococca; Onychocola canadensis;
Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces variotii;
Paracoccidioides brasiliensis; Penicillium marneffei; Penicillium
spp; Phialophora spp; Phialophora verrucosa; Phoma spp; Pichia
anomala; Pichia etchellsii; Pichia guilliermondii; Pichia ohmeri;
Pithomyces spp; Pneumocystis carinii; Pseudallescheria boydii;
Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor pusillus;
Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus; Rhizopus
oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis and Yarrowia
lipolytica); (b) contacting the cell with a substance to be tested
for the ability to inhibit fungal cell growth; (c) detecting
expression driven by the promoter or signal from the reporter in
the hybrid; (d) selecting the substance if it modulates expression
from the promoter or signal from the reporter. In an embodiment of
the invention, the method is carried out along with a
negative-control comprising (i) introducing the hybrid into a
suitable fungal cell; (ii) contacting the cell with a blank
substance which is known to not modulate expression from the
promoter or signal from the reporter in the hybrid; and (iii)
detecting expression driven by the promoter or signal from the
reporter in the hybrid; and (iv) comparing the expression level
from the promoter or signal from the reporter in the cell contacted
with the blank substance with the expression level from the
promoter or the signal from the reporter in the cell contacted with
the substance to be tested for the ability to inhibit fungal cell
growth. Embodiments of the invention also include methods wherein a
positive-control is carried out comprising (i) introducing the
hybrid into a suitable fungal cell; (ii) contacting the cell with a
positive-control which is known to modulate expression from the
promoter or signal from the reporter in the hybrid; (iii) detecting
expression driven by the promoter or signal from the reporter in
the hybrid; and (iv) comparing the expression level from the
promoter or signal from the reporter in the cell contacted with the
positive-control substance with the expression level from the
promoter or signal from the reporter in the cell contacted with the
substance to be tested for the ability to inhibit fungal cell
growth.
[0010] The present invention further comprises a method for
identifying a substance that inhibits fungal cell growth comprising
(a) introducing, into a suitable fungal cell (e.g., Absidia
corymbifera; Absidia spp; Acremonium spp; Ajellomyces capsulatus;
Ajellomyces dermatitidis; Alternaria spp; Aphanoascus fulvescens;
Apophysomyces spp; Arthroderma benhamiae; Arthroderma falvum;
Arthroderma gypseum; Arthroderma incurvatum; Arthroderma otae;
Arthroderma vanbreuseghemii; Aspergillus flavus; Aspergillus
fumigatus; Aspergillus glaucus; Aspergillus nidulans; Aspergillus
niger; Aspergillus oryzae; Aspergillus spp; Aspergillus sydowi;
Aspergillus terreus; Aspergillus ustus; Aspergillus versicolor;
Aureobasidium pullulans; Basidiomycetes; Beauveria spp; Bipolaris
hawaiiensis; Bipolaris spicifera; Bipolaris spp; Bjerkandera
adusta; Blastomyces dermatitidis; Blastoschizomyces capitatus;
Candida albicans; Candida beigelii; Candida colluculosa; Candida
dubliniensis; Candida dubliniensis; Candida famata; Candida famata;
Candida glabrata; Candida guilliermondii; Candida haemulonii;
Candida holmii; Candida inconspicua; Candida intermedia; Candida
keyfyr; Candida krusei; Candida krusei; Candida lambica; Candida
lipolytica; Candida lusitaniae; Candida maris; Candida melibiosica;
Candida norvegensis; Candida parapsilosis; Candida parapsilosis;
Candida pelliculosa; Candida pelliculosa; Candida pseudotropicalis;
Candida pulcherrima; Candida rugosa; Candida sake; Candida
sphaerica; Candida spp; Candida stellatoidea; Candida tropicalis;
Candida tropicalis; Candida viswanathii; Candida zeylanoides;
Chrysosporium spp; Cladophialophora bantiana; Cladophialophora
carrionii; Cladosporium spp; Coccidioides immitis; Cokeromyces
recurvatus; Coprinus spp; Cryptococcus albidus; Cryptococcus
gattii; Cryptococcus laurentii; Cryptococcus neoformans;
Cunninghamella bertholletiae; Cunninghamella spp; Curvularia
lunata; Curvularia spp; Dekkera bruxellensis; Epidermophyton
floccosum; Epidermophyton floccosum; Exophiala dermatitidis;
Exophiala jeanselmei; Exophiala moniliae; Exserohilum rostratum;
Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium dimerum;
Fusarium moniliforme; Fusarium oxysporum; Fusarium proliferatum;
Fusarium solani; Fusarium spp; Geotrichum candidum; Geotrichum spp;
Histoplasma capsulatum; Hortaea werneckii; Issatschenkia
orientalis; Kluveromyces lactis; Kluyveromyces marxianus; Madurella
grisae; Malassezia furfur; Malassezia globosa; Malassezia obtusa;
Malassezia pachydermatis; Malassezia restricta; Malassezia
slooffiae; Malassezia sympodialis; Metarrhizium anisopliae;
Microsporum audouinii; Microsporum canis; Microsporum fulvum;
Microsporum gypseum; Microsporum persicolor; Mucor circinelloides;
Mucor hiemalis; Mucor racemosus; Mucor rouxii; Mucor spp;
Nattrassia mangiferae; Nectria haematococca; Onychocola canadensis;
Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces variotii;
Paracoccidioides brasiliensis; Penicillium marneffei; Penicillium
spp; Phialophora spp; Phialophora verrucosa; Phoma spp; Pichia
anomala; Pichia etchellsii; Pichia guilliermondii; Pichia ohmeri;
Pithomyces spp; Pneumocystis carinii; Pseudallescheria boydii;
Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor pusillus;
Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus; Rhizopus
oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis or Yarrowia
lipolytica), a first hybrid comprising a first promoter selected
from the group consisting of S. cerevisiae YOR387C Promoter, S.
cerevisiae YMR175W Promoter, S. cerevisiae YFR026C Promoter, S.
cerevisiae YJL153C Promoter, S. cerevisiae YPL033C Promoter, S.
cerevisiae YOL058W Promoter, S. cerevisiae YOR255W Promoter, S.
cerevisiae YDR250C Promoter, S. cerevisiae YDR446W Promoter, S.
cerevisiae YDR536W Promoter, S. cerevisiae YOR255W Promoter, S.
cerevisiae YDL243C Promoter S. cerevisiae YDR256C Promoter, S.
cerevisiae YFL020C Promoter, S. cerevisiae YPL205C Promoter, S.
cerevisiae YGL205W Promoter, S. cerevisiae YGL117W Promoter, S.
cerevisiae YHR029C Promoter, S. cerevisiae YML116W Promoter S.
cerevisiae YJR109C Promoter, S. cerevisiae YCL030C Promoter, S.
cerevisiae YLR338W Promoter, S. cerevisiae YIL066W-A Promoter, S.
cerevisiae YNL093W Promoter, S. cerevisiae YLR381W Promoter, S.
cerevisiae YKL159C Promoter, S. cerevisiae YIL058W Promoter, S.
cerevisiae YKR037C Promoter, S. cerevisiae YNL279W Promoter, S.
cerevisiae YOR032C Promoter, S. cerevisiae YML058W Promoter, S.
cerevisiae YMR303C Promoter, S. cerevisiae YJL153C Promoter S.
cerevisiae YLR092W Promoter, S. cerevisiae YFL052W Promoter, S.
cerevisiae YIR017C Promoter, S. cerevisiae YLL062C Promoter, S.
cerevisiae YMR323W Promoter S. cerevisiae YPL171C Promoter, S.
cerevisiae YER065C Promoter, S. cerevisiae YDR114C Promoter and S.
cerevisiae YBL065W Promoter (e.g., SEQ ID NOs: 1-41 or 68);
operably linked to a first reporter (e.g., S. cerevisiae ADE2, S.
cerevisiae LYS2, S. cerevisiae TRP1, S. cerevisiae LEU2, S.
cerevisiae URA3, S. cerevisiae HIS3, Aequorea victoria GFP mutant
3, Renilla luciferase, Photinus pyralis luciferase, Photinus
pyralis luciferase slk mutant, Vibrio fischeri luxA, Vibrio
fischeri luxB, Vibrio fischeri luxC, Vibrio fischeri luxD, Vibrio
fischeri luxE, Vibrio fischeri luxAB, Vibrio fischeri luxCDABE,
Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio harveyi luxC,
Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio harveyi luxAB,
Vibrio harveyi luxCDABE, Photorhabdus luminscens LuxA, Photorhabdus
luminscens LuxB, Photorhabdus luminscens LuxC, Photorhabdus
luminscens LuxD, Photorhabdus luminscens LuxE, Photorhabdus
luminscens LuxCDABE, E. coli lacZ, the Aequorea victoria Aequorin
gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS, CYH2 or CAN1 (e.g.,
SEQ ID NOs: 42-51)); and a second hybrid comprising a second
promoter selected from the group consisting of: S. cerevisiae
YHR082C Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae
YBR106W Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae
YPR108W Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae
YJL166W Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae
YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae
YHR012W Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae
YKL196C Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae
YLR427W Promoter and S. cerevisiae YLL020C Promoter (e.g., SEQ ID
NOs: 52-67); operably linked to a second reporter (e.g., S.
cerevisiae ADE2, S. cerevisiae LYS2, S. cerevisiae TRP1, S.
cerevisiae LEU2, S. cerevisiae URA3, S. cerevisiae HIS3, Aequorea
victoria GFP mutant 3, Renilla luciferase, Photinus pyralis
luciferase, Photinus pyralis luciferase slk mutant, Vibrio fischeri
luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio fischeri
luxD, Vibrio fischeri luxE, Vibrio fischeri luxAB, Vibrio fischeri
luxCDABE, Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio harveyi
luxC, Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio harveyi
luxAB, Vibrio harveyi luxCDABE, Photorhabdus luminscens LuxA,
Photorhabdus luminscens LuxB, Photorhabdus luminscens LuxC,
Photorhabdus luminscens LuxD, Photorhabdus luminscens LuxE,
Photorhabdus luminscens LuxCDABE, E. coli lacZ, the Aequorea
victoria Aequorin gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS,
CYH2 and CAN1 (e.g., SEQ ID NOs: 42-51)); wherein said first and
second reporters are different; (b) contacting the cell with a
substance to be tested for the ability to inhibit fungal cell
growth; (c) detecting expression driven by the first and second
promoters or signal from the reporters; and (d) selecting the
substance if it causes the first promoter expression level or
reporter signal to increase or decrease in relation to the
expression level from the second promoter or signal from the second
reporter. In an embodiment of the present invention, the method is
carried out along with a negative-control comprising (i)
introducing said first and second hybrids into a suitable fungal
cell; (ii) contacting the cell with a blank substance which is
known to not modulate expression from the promoters or signal from
the reporters in the hybrids; and (iii) detecting expression driven
by the promoters or signal from the reporters in the hybrids; (iv)
comparing the expression level from the first promoter or signal
from the first reporter in relation to the expression level from
the second promoter or signal from the second reporter in the cell
contacted with the blank substance with the expression level from
the first promoter or signal from the first reporter in relation to
the expression level from the second promoter or signal from the
second reporter in the cell contacted with the substance to be
tested for the ability to inhibit fungal cell growth. In an
embodiment of the invention, the method is carried out along with a
positive-control comprising: (i) introducing said first and second
hybrids into a suitable fungal cell; (ii) contacting the cell with
a positive-control substance which is known to modulate expression
from the promoters or signal from the reporters in the hybrids;
(iii) detecting expression driven by the promoters or signal from
the reporters in the hybrids; and (iv) comparing the expression
level from the first promoter or signal from the first reporter in
relation to the expression level from the second promoter or signal
from the second reporter in the cell contacted with the
positive-control substance with the expression level from the first
promoter or signal from the first reporter in relation to the
expression level from the second promoter or signal from the second
reporter in the cell contacted with the substance to be tested for
the ability to inhibit fungal cell growth.
[0011] The present invention also provides a method for determining
whether a substance inhibits growth of a fungal cell comprising (a)
introducing a hybrid comprising a promoter selected from the group
consisting of S. cerevisiae YHR082C Promoter, S. cerevisiae YDL126C
Promoter S. cerevisiae YBR106W Promoter, S. cerevisiae YLL043W
Promoter, S. cerevisiae YPR108W Promoter, S. cerevisiae YIL041W
Promoter, S. cerevisiae YJL166W Promoter S. cerevisiae YDR165W
Promoter, S. cerevisiae YLR330 W Promoter, S. cerevisiae YDL058W
Promoter, S. cerevisiae YHR012W Promoter, S. cerevisiae YKL053W
Promoter, S. cerevisiae YKL196C Promoter, S. cerevisiae YDL103C
Promoter S. cerevisiae YLR427W Promoter and S. cerevisiae YLL020C
Promoter (e.g., SEQ ID NOs: 52-67); operably linked to a reporter
(e.g., S. cerevisiae ADE2, S. cerevisiae LYS2, S. cerevisiae TRP1,
S. cerevisiae LEU2, S. cerevisiae URA3, S. cerevisiae HIS3,
Aequorea victoria GFP mutant 3, Renilla luciferase, Photinus
pyralis luciferase, Photinus pyralis luciferase slk mutant, Vibrio
fischeri luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio
fischeri luxD, Vibrio fischeri luxE, Vibrio fischeri luxAB, Vibrio
fischeri luxCDABE, Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio
harveyi luxC, Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio
harveyi luxAB, Vibrio harveyi luxCDABE, Photorhabdus luminscens
LuxA, Photorhabdus luminscens LuxB, Photorhabdus luminscens LuxC,
Photorhabdus luminscens LuxD, Photorhabdus luminscens LuxE,
Photorhabdus luminscens LuxCDABE, E. coli lacZ, the Aequorea
victoria Aequorin gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS,
CYH2 or CAN1 (e.g., SEQ ID NOs: 42-51)) into a suitable fungal cell
(e.g., Absidia corymbifera; Absidia spp; Acremonium spp;
Ajellomyces capsulatus; Ajellomyces dermatitidis; Alternaria spp;
Aphanoascus fulvescens; Apophysomyces spp; Arthroderma benhamiae;
Arthroderma falvum; Arthroderma gypseum; Arthroderma incurvatum;
Arthroderma otae; Arthroderma vanbreuseghemii; Aspergillus flavus;
Aspergillus fumigatus; Aspergillus glaucus; Aspergillus nidulans;
Aspergillus niger; Aspergillus oryzae; Aspergillus spp; Aspergillus
sydowi; Aspergillus terreus; Aspergillus ustus; Aspergillus
versicolor; Aureobasidium pullulans; Basidiomycetes; Beauveria spp;
Bipolaris hawaiiensis; Bipolaris spicifera; Bipolaris spp;
Bjerkandera adusta; Blastomyces dermatitidis; Blastoschizomyces
capitatus; Candida albicans; Candida beigelii; Candida colluculosa;
Candida dubliniensis; Candida dubliniensis; Candida famata; Candida
famata; Candida glabrata; Candida guilliermondii; Candida
haemulonii; Candida holmii; Candida inconspicua; Candida
intermedia; Candida keyfyr; Candida krusei; Candida krusei; Candida
lambica; Candida lipolytica; Candida lusitaniae; Candida maris;
Candida melibiosica; Candida norvegensis; Candida parapsilosis;
Candida parapsilosis; Candida pelliculosa; Candida pelliculosa;
Candida pseudotropicalis; Candida pulcherrima; Candida rugosa;
Candida sake; Candida sphaerica; Candida spp; Candida stellatoidea;
Candida tropicalis; Candida tropicalis; Candida viswanathii;
Candida zeylanoides; Chrysosporium spp; Cladophialophora bandana;
Cladophialophora carrionii; Cladosporium spp; Coccidioides immitis;
Cokeromyces recurvatus; Coprinus spp; Cryptococcus albidus;
Cryptococcus gattii; Cryptococcus laurentii; Cryptococcus
neoformans; Cunninghamella bertholletiae; Cunninghamella spp;
Curvularia lunata; Curvularia spp; Dekkera bruxellensis;
Epidermophyton floccosum; Epidermophyton floccosum; Exophiala
dermatitidis; Exophiala jeanselmei; Exophiala moniliae; Exserohilum
rostratum; Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium
dimerum; Fusarium moniliforme; Fusarium oxysporum; Fusarium
proliferatum; Fusarium solani; Fusarium spp; Geotrichum candidum;
Geotrichum spp; Histoplasma capsulatum; Hortaea werneckii;
Issatschenkia orientalis; Kluveromyces lactis; Kluyveromyces
marxianus; Madurella grisae; Malassezia furfur; Malassezia globosa;
Malassezia obtusa; Malassezia pachydermatis; Malassezia restricta;
Malassezia slooffiae; Malassezia sympodialis; Metarrhizium
anisopliae; Microsporum audouinii; Microsporum canis; Microsporum
fulvum; Microsporum gypseum; Microsporum persicolor; Mucor
circinelloides; Mucor hiemalis; Mucor racemosus; Mucor rouxii;
Mucor spp; Nattrassia mangiferae; Nectria haematococca; Onychocola
canadensis; Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces
variotii; Paracoccidioides brasiliensis; Penicillium marneffei;
Penicillium spp; Phialophora spp; Phialophora verrucosa; Phoma spp;
Pichia anomala; Pichia etchellsii; Pichia guilliermondii; Pichia
ohmeri; Pithomyces spp; Pneumocystis carinii; Pseudallescheria
boydii; Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor
pusillus; Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus;
Rhizopus oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis or Yarrowia
lipolytica); (b) contacting the cell with a substance to be tested
for the ability to inhibit growth of the cell; (c) detecting signal
from the reporter; and (d) selecting the substance if it causes a
reduction in the signal from the reporter and/or calculating the IC
of the substance. In an embodiment of the present invention, the
method is carried out along with a negative-control comprising: (i)
introducing the hybrid into a suitable fungal cell; (ii) contacting
the cell with a blank substance which is known to not inhibit
fungal cell growth; (iii) detecting signal from the reporter in the
hybrid; and (iv) comparing the signal from the reporter in the cell
contacted with the blank substance with the signal from the
reporter in the cell contacted with the substance to be tested for
the ability to inhibit fungal cell growth. In an embodiment of the
invention, the method is carried out along with a positive-control
comprising (i) introducing the hybrid into a suitable fungal cell;
(ii) contacting the cell with a positive-control substance which is
known to inhibit fungal cell growth; (iii) detecting signal from
the reporter in the hybrid; and (iv) comparing the signal from the
reporter in the cell contacted with the positive-control substance
with the signal from the reporter in the cell contacted with the
substance to be tested for the ability to inhibit fungal cell
growth.
[0012] The present invention further provides a method for
determining whether a substance inhibits growth of a fungal cell
comprising (a) introducing a first hybrid comprising a first
promoter selected from the group consisting of S. cerevisiae
YHR082C Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae
YBR106W Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae
YPR108W Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae
YJL166W Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae
YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae
YHR012W Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae
YKL196C Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae
YLR427W Promoter and S. cerevisiae YLL020C Promoter (e.g., SEQ ID
NOs: 52-67); operably linked to a first reporter (e.g., S.
cerevisiae ADE2, S. cerevisiae LYS2, S. cerevisiae TRP1, S.
cerevisiae LEU2, S. cerevisiae URA3, S. cerevisiae HIS3, Aequorea
victoria GFP mutant 3, Renilla luciferase, Photinus pyralis
luciferase, Photinus pyralis luciferase slk mutant, Vibrio fischeri
luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio fischeri
luxD, Vibrio fischeri luxE, Vibrio fischeri luxAB, Vibrio fischeri
luxCDABE, Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio harveyi
luxC, Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio harveyi
luxAB, Vibrio harveyi luxCDABE, Photorhabdus luminscens LuxA,
Photorhabdus luminscens LuxB, Photorhabdus luminscens LuxC,
Photorhabdus luminscens LuxD, Photorhabdus luminscens LuxE,
Photorhabdus luminscens LuxCDABE, E. coli lacZ, the Aequorea
victoria Aequorin gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS,
CYH2 or CAN1 (e.g., SEQ ID NOs: 42-51)) and a second hybrid
comprising a second promoter selected from the group consisting of
S. cerevisiae YHR082C Promoter, S. cerevisiae YDL126C Promoter S.
cerevisiae YBR106W Promoter, S. cerevisiae YLL043W Promoter, S.
cerevisiae YPR108W Promoter, S. cerevisiae YIL041W Promoter, S.
cerevisiae YIL166W Promoter S. cerevisiae YDR165W Promoter, S.
cerevisiae YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S.
cerevisiae YHR012W Promoter, S. cerevisiae YKL053W Promoter, S.
cerevisiae YKL196C Promoter, S. cerevisiae YDL103C Promoter S.
cerevisiae YLR427W Promoter and S. cerevisiae YLL020C Promoter
(e.g., SEQ ID NOs: 52-67); operably linked to a second reporter
(e.g., S. cerevisiae ADE2, S. cerevisiae LYS2, S. cerevisiae TRP1,
S. cerevisiae LEU2, S. cerevisiae URA3, S. cerevisiae HIS3,
Aequorea victoria GFP mutant 3, Renilla luciferase, Photinus
pyralis luciferase, Photinus pyralis luciferase slk mutant, Vibrio
fischeri luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio
fischeri luxD, Vibrio fischeri luxE, Vibrio fischeri luxAB, Vibrio
fischeri luxCDABE, Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio
harveyi luxC, Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio
harveyi luxAB, Vibrio harveyi luxCDABE, Photorhabdus luminscens
LuxA, Photorhabdus luminscens LuxB, Photorhabdus luminscens LuxC,
Photorhabdus luminscens LuxD, Photorhabdus luminscens LuxE,
Photorhabdus luminscens LuxCDABE, E. coli lacZ, the Aequorea
victoria Aequorin gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS,
CYH2 or CAN1 (e.g., SEQ ID NOs: 42-51)) into a suitable fungal cell
(e.g., Absidia corymbifera; Absidia spp; Acremonium spp;
Ajellomyces capsulatus; Ajellomyces dermatitidis; Alternaria spp;
Aphanoascus fulvescens; Apophysomyces spp; Arthroderma benhamiae;
Arthroderma fulvum; Arthroderma gypseum; Arthroderma incurvatum;
Arthroderma otae; Arthroderma vanbreuseghemii; Aspergillus flavus;
Aspergillus fumigatus; Aspergillus glaucus; Aspergillus nidulans;
Aspergillus niger; Aspergillus oryzae; Aspergillus spp; Aspergillus
sydowi; Aspergillus terreus; Aspergillus ustus; Aspergillus
versicolor; Aureobasidium pullulans; Basidiomycetes; Beauveria spp;
Bipolaris hawaiiensis; Bipolaris spicifera; Bipolaris spp;
Bjerkandera adusta; Blastomyces dermatitidis; Blastoschizomyces
capitatus; Candida albicans; Candida beigelii; Candida colluculosa;
Candida dubliniensis; Candida dubliniensis; Candida famata; Candida
famata; Candida glabrata; Candida guilliermondii; Candida
haemulonii; Candida holmii; Candida inconspicua; Candida
intermedia; Candida keyfyr; Candida krusei; Candida krusei; Candida
lambica; Candida lipolytica; Candida lusitaniae; Candida maris;
Candida melibiosica; Candida norvegensis; Candida parapsilosis;
Candida parapsilosis; Candida pelliculosa; Candida pelliculosa;
Candida pseudotropicalis; Candida pulcherrima; Candida rugosa;
Candida sake; Candida sphaerica; Candida spp; Candida stellatoidea;
Candida tropicalis; Candida tropicalis; Candida viswanathii;
Candida zeylanoides; Chrysosporium spp; Cladophialophora bantiana;
Cladophialophora carrionii; Cladosporium spp; Coccidioides immitis;
Cokeromyces recurvatus; Coprinus spp; Cryptococcus albidus;
Cryptococcus gattii; Cryptococcus laurentii; Cryptococcus
neoformans; Cunninghamella bertholletiae; Cunninghamella spp;
Curvularia lunata; Curvularia spp; Dekkera bruxellensis;
Epidermophyton floccosum; Epidermophyton floccosum; Exophiala
dermatitidis; Exophiala jeanselmei; Exophiala moniliae; Exserohilum
rostratum; Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium
dimerum; Fusarium moniliforme; Fusarium oxysporum; Fusarium
proliferatum; Fusarium solani; Fusarium spp; Geotrichum candidum;
Geotrichum spp; Histoplasma capsulatum; Hortaea werneckii;
Issatschenkia orientalis; Kluveromyces lactis; Kluyveromyces
marxianus; Madurella grisae; Malassezia furfur; Malassezia globosa;
Malassezia obtusa; Malassezia pachydermatis; Malassezia restricta;
Malassezia slooffiae; Malassezia sympodialis; Metarrhizium
anisopliae; Microsporum audouinii; Microsporum canis; Microsporum
fulvum; Microsporum gypseum; Microsporum persicolor; Mucor
circinelloides; Mucor hiemalis; Mucor racemosus; Mucor rouxii;
Mucor spp; Nattrassia mangiferae; Nectria haematococca; Onychocola
canadensis; Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces
variotii; Paracoccidioides brasiliensis; Penicillium marneffei;
Penicillium spp; Phialophora spp; Phialophora verrucosa; Phoma spp;
Pichia anomala; Pichia etchellsii; Pichia guilliermondii; Pichia
ohmeri; Pithomyces spp; Pneumocystis carinii; Pseudallescheria
boydii; Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor
pusillus; Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus;
Rhizopus oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis or Yarrowia
lipolytica); wherein said first and second promoter are different
and wherein said first and second reporter are different; (b)
contacting the cell with a substance to be tested for the ability
to inhibit growth of the cell; (c) detecting signal from the
reporters; and (d) selecting the substance if it causes a reduction
in the signal from both the first and second reporter and/or
calculating the IC of the substance. In an embodiment of the
present invention, the method is carried out along with a
negative-control comprising (i) introducing the first and second
hybrids into a suitable fungal cell; (ii) contacting the cell with
a blank substance which is known to not inhibit fungal cell growth;
(iii) detecting signal from the first and second reporters in the
hybrids; and (iv) comparing the signal from the first and second
reporters in the cell contacted with the blank substance with the
signal from the first and second reporters in the cell contacted
with the substance to be tested for the ability to inhibit fungal
cell growth. In another embodiment of the invention, the method is
carried out along with a positive-control comprising (i)
introducing the first and second hybrids into a suitable fungal
cell; (ii) contacting the cell with a positive control substance
which is known to inhibit fungal cell growth; (iii) detecting
signal from the first and second promoters in the hybrids; and (iv)
comparing the signal from the first and second reporters in the
cell contacted with the positive control substance with the signal
from the first and second reporters in the cell contacted with the
substance to be tested for the ability to inhibit fungal cell
growth.
[0013] The present invention also provides a method for determining
whether a substance inhibits growth of a malignant cell comprising
(a) introducing, into a suitable fungal cell (e.g., Absidia
corymbifera; Absidia spp; Acremonium spp; Ajellomyces capsulatus;
Ajellomyces dermatitidis; Alternaria spp; Aphanoascus fulvescens;
Apophysomyces spp; Arthroderma benhamiae; Arthroderma fulvum;
Arthroderma gypseum; Arthroderma incurvatum; Arthroderma otae;
Arthroderma vanbreuseghemii; Aspergillus flavus; Aspergillus
fumigatus; Aspergillus glaucus; Aspergillus nidulans; Aspergillus
niger; Aspergillus oryzae; Aspergillus spp; Aspergillus sydowi;
Aspergillus terreus; Aspergillus ustus; Aspergillus versicolor;
Aureobasidium pullulans; Basidiomycetes; Beauveria spp; Bipolaris
hawaiiensis; Bipolaris spicifera; Bipolaris spp; Bjerkandera
adusta; Blastomyces dermatitidis; Blastoschizomyces capitatus;
Candida albicans; Candida beigelii; Candida colluculosa; Candida
dubliniensis; Candida dubliniensis; Candida famata; Candida famata;
Candida glabrata; Candida guilliermondii; Candida haemulonii;
Candida holmii; Candida inconspicua; Candida intermedia; Candida
keyfyr; Candida krusei; Candida krusei; Candida lambica; Candida
lipolytica; Candida lusitaniae; Candida maris; Candida melibiosica;
Candida norvegensis; Candida parapsilosis; Candida parapsilosis;
Candida pelliculosa; Candida pelliculosa; Candida pseudotropicalis;
Candida pulcherrima; Candida rugosa; Candida sake; Candida
sphaerica; Candida spp; Candida stellatoidea; Candida tropicalis;
Candida tropicalis; Candida viswanathii; Candida zeylanoides;
Chrysosporium spp; Cladophialophora bantiana; Cladophialophora
carrionii; Cladosporium spp; Coccidioides immitis; Cokeromyces
recurvatus; Coprinus spp; Cryptococcus albidus; Cryptococcus
gattii; Cryptococcus laurentii; Cryptococcus neoformans;
Cunninghamella bertholletiae; Cunninghamella spp; Curvularia
lunata; Curvularia spp; Dekkera bruxellensis; Epidermophyton
floccosum; Epidermophyton floccosum; Exophiala dermatitidis;
Exophiala jeanselmei; Exophiala moniliae; Exserohilum rostratum;
Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium dimerum;
Fusarium moniliforme; Fusarium oxysporum; Fusarium proliferatum;
Fusarium solani; Fusarium spp; Geotrichum candidum; Geotrichum spp;
Histoplasma capsulatum; Hortaea werneckii; Issatschenkia
orientalis; Kluveromyces lactis; Kluyveromyces marxianus; Madurella
grisae; Malassezia furfur; Malassezia globosa; Malassezia obtusa;
Malassezia pachydermatis; Malassezia restricta; Malassezia
slooffiae; Malassezia sympodialis; Metarrhizium anisopliae;
Microsporum audouinii; Microsporum canis; Microsporum fulvum;
Microsporum gypseum; Microsporum persicolor; Mucor circinelloides;
Mucor hiemalis; Mucor racemosus; Mucor rouxii; Mucor spp;
Nattrassia mangiferae; Nectria haematococca; Onychocola canadensis;
Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces variotii;
Paracoccidioides brasiliensis; Penicillium marneffei; Penicillium
spp; Phialophora spp; Phialophora verrucosa; Phoma spp; Pichia
anomala; Pichia etchellsii; Pichia guilliermondii; Pichia ohmeri;
Pithomyces spp; Pneumocystis carinii; Pseudallescheria boydii;
Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor pusillus;
Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus; Rhizopus
oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis or Yarrowia
lipolytica), a first hybrid comprising a first promoter selected
from the group consisting of: S. cerevisiae YLR338W, S. cerevisiae
YIL066W, S. cerevisiae YNL093W, S. cerevisiae YDR250C, S.
cerevisiae YLR381W, S. cerevisiae YBL065W, S. cerevisiae YMR323W
and S. cerevisiae YPL171C (e.g., SEQ ID NOs: 8, 22-25, 38, 39 and
68); operably linked to a first reporter (e.g., S. cerevisiae ADE2,
S. cerevisiae LYS2, S. cerevisiae TRP1, S. cerevisiae LEU2, S.
cerevisiae URA3, S. cerevisiae HIS3, Aequorea victoria GFP mutant
3, Renilla luciferase, Photinus pyralis luciferase, Photinus
pyralis luciferase slk mutant, Vibrio fischeri luxA, Vibrio
fischeri luxB, Vibrio fischeri luxC, Vibrio fischeri luxD, Vibrio
fischeri luxE, Vibrio fischeri luxAB, Vibrio fischeri luxCDABE,
Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio harveyi luxC,
Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio harveyi luxAB,
Vibrio harveyi luxCDABE, Photorhabdus luminscens LuxA, Photorhabdus
luminscens LuxB, Photorhabdus luminscens LuxC, Photorhabdus
luminscens LuxD, Photorhabdus luminscens LuxE, Photorhabdus
luminscens LuxCDABE, E. coli lacZ, the Aequorea victoria Aequorin
gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS, CYH2 or CAN1 (e.g.,
SEQ ID NO: 42-51)); and a second hybrid comprising a second
promoter selected from the group consisting of: S. cerevisiae
YHR082C Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae
YBR106W Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae
YPR108W Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae
YJL166W Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae
YLR330 W Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae
YHR012W Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae
YKL196C Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae
YLR427W Promoter and S. cerevisiae YLL020C Promoter (e.g., SEQ ID
NOs: 52-67); operably linked to a second reporter (e.g., S.
cerevisiae ADE2, S. cerevisiae LYS2, S. cerevisiae TRP1, S.
cerevisiae LEU2, S. cerevisiae URA3, S. cerevisiae HIS3, Aequorea
victoria GFP mutant 3, Renilla luciferase, Photinus pyralis
luciferase, Photinus pyralis luciferase slk mutant, Vibrio fischeri
luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio fischeri
luxD, Vibrio fischeri luxE, Vibrio fischeri luxAB, Vibrio fischeri
luxCDABE, Vibrio harveyi luxA, Vibrio harveyi luxB, Vibrio harveyi
luxC, Vibrio harveyi luxD, Vibrio harveyi luxE, Vibrio harveyi
luxAB, Vibrio harveyi luxCDABE, Photorhabdus luminscens LuxA,
Photorhabdus luminscens LuxB, Photorhabdus luminscens LuxC,
Photorhabdus luminscens LuxD, Photorhabdus luminscens LuxE,
Photorhabdus luminscens LuxCDABE, E. coli lacZ, the Aequorea
victoria Aequorin gene, KanMX, pat1, nat1, hph, CAT, Sh Ble, GUS,
CYH2 or CAN1 (e.g., SEQ ID NO: 42-51)); (b) contacting the cell
with a substance to be tested for the ability to inhibit growth of
a malignant cell; (c) detecting expression driven by the first and
second promoters; and (d) selecting the substance if it causes the
expression level from the first promoter to increase or decrease in
relation to the expression level from the second promoter. In an
embodiment of the invention, the method is carried out along with a
negative-control comprising (i) introducing said first and second
hybrids into a suitable fungal cell; (ii) contacting the cell with
a blank substance which is known to not modulate expression from
the promoters in the hybrids; and (iii) detecting expression driven
by the promoters in the hybrids; (iv) comparing the expression
level from the first promoter in relation to the expression level
from the second promoter in the cell contacted with the blank
substance with the expression level from the first promoter in
relation to the expression level from the second promoter in the
cell contacted with the substance to be tested for the ability to
inhibit malignant cell growth. In an embodiment of the invention,
the method is carried out along with a positive-control comprising
(i) introducing said first and second hybrids into a suitable
fungal cell; (ii) contacting the cell with a positive-control
substance which is known to modulate expression from the promoters
in the hybrids; (iii) detecting expression driven by the promoters
in the hybrids; and (iv) comparing the expression level from the
first promoter in relation to the expression level from the second
promoter in the cell contacted with the positive-control substance
with the expression level from the first promoter in relation to
the expression level from the second promoter in the cell contacted
with the substance to be tested for the ability to inhibit
malignant cell growth.
DETAILED DESCRIPTION OF THE INVENTION
[0014] Each of the following isolated polynucleotides (with the
exception SEQ ID NOs: 42-51), optionally fused to a reporter, are
part of the present invention along with the various methods of use
thereof which are described herein.
TABLE-US-00001 TABLE 1 Polynucleic acids of the invention. SEQUENCE
SEQUENCE IDENTIFIER S. cerevisiae YOR387C Promoter SEQ ID NO: 1 S.
cerevisiae YMR175W Promoter SEQ ID NO: 2 S. cerevisiae YFR026C
Promoter SEQ ID NO: 3 S. cerevisiae YJL153C Promoter SEQ ID NO: 4
S. cerevisiae YPL033C Promoter SEQ ID NO: 5 S. cerevisiae YOL058W
Promoter SEQ ID NO: 6 S. cerevisiae YOR255W Promoter SEQ ID NO: 7
S. cerevisiae YDR250C Promoter SEQ ID NO: 8 S. cerevisiae YDR446W
Promoter SEQ ID NO: 9 S. cerevisiae YDR536W Promoter SEQ ID NO: 10
S. cerevisiae YOR255W Promoter SEQ ID NO: 11 S. cerevisiae YDL243C
Promoter SEQ ID NO: 12 S. cerevisiae YDR256C Promoter SEQ ID NO: 13
S. cerevisiae YFL020C Promoter SEQ ID NO: 14 S. cerevisiae YPL205C
Promoter SEQ ID NO: 15 S. cerevisiae YGL205W Promoter SEQ ID NO: 16
S. cerevisiae YGL117W Promoter SEQ ID NO: 17 S. cerevisiae YHR029C
Promoter SEQ ID NO: 18 S. cerevisiae YML116W Promoter SEQ ID NO: 19
S. cerevisiae YJR109C Promoter SEQ ID NO: 20 S. cerevisiae YCL030C
Promoter SEQ ID NO: 21 S. cerevisiae YLR338W Promoter SEQ ID NO: 22
S. cerevisiae YIL066W-A Promoter SEQ ID NO: 23 S. cerevisiae
YNL093W Promoter SEQ ID NO: 24 S. cerevisiae YLR381W Promoter SEQ
ID NO: 25 S. cerevisiae YKL159C Promoter SEQ ID NO: 26 S.
cerevisiae YIL058W Promoter SEQ ID NO: 27 S. cerevisiae YKR037C
Promoter SEQ ID NO: 28 S. cerevisiae YNL279W Promoter SEQ ID NO: 29
S. cerevisiae YOR032C Promoter SEQ ID NO: 30 S. cerevisiae YML058W
Promoter SEQ ID NO: 31 S. cerevisiae YMR303C Promoter SEQ ID NO: 32
S. cerevisiae YJL153C Promoter SEQ ID NO: 33 S. cerevisiae YLR092W
Promoter SEQ ID NO: 34 S. cerevisiae YFL052W Promoter SEQ ID NO: 35
S. cerevisiae YIR017C Promoter SEQ ID NO: 36 S. cerevisiae YLL062C
Promoter SEQ ID NO: 37 S. cerevisiae YMR323W Promoter SEQ ID NO: 38
S. cerevisiae YPL171C Promoter SEQ ID NO: 39 S. cerevisiae YER065C
Promoter SEQ ID NO: 40 S. cerevisiae YDR114C Promoter SEQ ID NO: 41
S. cerevisiae ADE2 SEQ ID NO: 42 S. cerevisiae LYS2 SEQ ID NO: 43
S. cerevisiae TRP1 SEQ ID NO: 44 S. cerevisiae LEU2 SEQ ID NO: 45
S. cerevisiae URA3 SEQ ID NO: 46 S. cerevisiae HIS3 SEQ ID NO: 47
Aequorea victoria GFP mutant 3 SEQ ID NO: 48 Renilla luciferase SEQ
ID NO: 49 Photinus pyralis luciferase SEQ ID NO: 50 E. coli lacZ
SEQ ID NO: 51 S. cerevisiae YHR082C Promoter SEQ ID NO: 52 S.
cerevisiae YDL126C Promoter SEQ ID NO: 53 S. cerevisiae YBR106W
Promoter SEQ ID NO: 54 S. cerevisiae YLL043W Promoter SEQ ID NO: 55
S. cerevisiae YPR108W Promoter SEQ ID NO: 56 S. cerevisiae YIL041W
Promoter SEQ ID NO: 57 S. cerevisiae YJL166W Promoter SEQ ID NO: 58
S. cerevisiae YDR165W Promoter SEQ ID NO: 59 S. cerevisiae YLR330W
Promoter SEQ ID NO: 60 S. cerevisiae YDL058W Promoter SEQ ID NO: 61
S. cerevisiae YHR012W Promoter SEQ ID NO: 62 S. cerevisiae YKL053W
Promoter SEQ ID NO: 63 S. cerevisiae YKL196C Promoter SEQ ID NO: 64
S. cerevisiae YDL103C Promoter SEQ ID NO: 65 S. cerevisiae YLR427W
Promoter SEQ ID NO: 66 S. cerevisiae YLL020C Promoter SEQ ID NO: 67
S. cerevisiae YBL065W Promoter SEQ ID NO: 68 PR131 Primer SEQ ID
NO: 69 PR132 Primer SEQ ID NO: 70 PR161 Primer SEQ ID NO: 71 PR
162Primer SEQ ID NO: 72 PHO 5 signal sequence SEQ ID NO: 73 PR172
Primer SEQ ID NO: 74 PR173B Primer SEQ ID NO: 75 PR207 Primer SEQ
ID NO: 76 PR208 Primer SEQ ID NO: 77 PR213 Primer SEQ ID NO: 78
PR214Primer SEQ ID NO: 79 PR167 Primer SEQ ID NO: 80 PR168SLK
Primer SEQ ID NO: 81 PHO5 signal sequence SEQ ID NO: 82 PR174
Primer SEQ ID NO: 83 PR175B Primer SEQ ID NO: 84 pSPRT47 plasmid
sequence SEQ ID NO: 85 pSPRT50 plasmid sequence SEQ ID NO: 86
pSPRT190 plasmid sequence SEQ ID NO: 87 pSPRT192 plasmid sequence
SEQ ID NO: 88
[0015] Since the Saccharmomyces cerevisiae genome has been
sequenced, the promoter sequences identified in Table 1 can also be
found in publicly available databases including the Saccharomyces
Genome Database; Stanford University, Calif.
[0016] The term "fungal cell" or "fungi" or "fungus" includes, but
is not limited to, any of the following organisms: Absidia
corymbifera; Absidia spp; Acremonium spp; Ajellomyces capsulatus;
Ajellomyces dermatitidis; Alternaria spp; Aphanoascus fulvescens;
Apophysomyces spp; Arthroderma benhamiae; Arthroderma fulvum;
Arthroderma gypseum; Arthroderma incurvatum; Arthroderma otae;
Arthroderma vanbreuseghemii; Aspergillus flavus; Aspergillus
fumigatus; Aspergillus glaucus; Aspergillus nidulans; Aspergillus
niger; Aspergillus oryzae; Aspergillus spp; Aspergillus sydowi;
Aspergillus terreus; Aspergillus ustus; Aspergillus versicolor;
Aureobasidium pullulans; Basidiomycetes; Beauveria spp; Bipolaris
hawaiiensis; Bipolaris spicifera; Bipolaris spp; Bjerkandera
adusta; Blastomyces dermatitidis; Blastoschizomyces capitatus;
Candida albicans; Candida beigelii; Candida colluculosa; Candida
dubliniensis; Candida dubliniensis; Candida famata; Candida famata;
Candida glabrata; Candida guilliermondii; Candida haemulonii;
Candida holmii; Candida inconspicua; Candida intermedia; Candida
keyfyr; Candida krusei; Candida krusei; Candida lambica; Candida
lipolytica; Candida lusitaniae; Candida maris; Candida melibiosica;
Candida norvegensis; Candida parapsilosis; Candida parapsilosis;
Candida pelliculosa; Candida pelliculosa; Candida pseudotropicalis;
Candida pulcherrima; Candida rugosa; Candida sake; Candida
sphaerica; Candida spp; Candida stellatoidea; Candida tropicalis;
Candida tropicalis; Candida viswanathii; Candida zeylanoides;
Chrysosporium spp; Cladophialophora bantiana; Cladophialophora
carrionii; Cladosporium spp; Coccidioides immitis; Cokeromyces
recurvatus; Coprinus spp; Cryptococcus albidus; Cryptococcus
gattii; Cryptococcus laurentii; Cryptococcus neoformans;
Cunninghamella bertholletiae; Cunninghamella spp; Curvularia
lunata; Curvularia spp; Dekkera bruxellensis; Epidermophyton
floccosum; Epidermophyton floccosum; Exophiala dermatitidis;
Exophiala jeanselmei; Exophiala moniliae; Exserohilum rostratum;
Filobasidiella neoformans; Fonsecaea pedrosoi; Fusarium dimerum;
Fusarium moniliforme; Fusarium oxysporum; Fusarium prolaferatum;
Fusarium solani; Fusarium spp; Geotrichum candidum; Geotrichum spp;
Histoplasma capsulatum; Hortaea werneckii; Issatschenkia
orientalis; Kluveromyces lactis; Kluyveromyces marxianus; Madurella
grisae; Malassezia furfur; Malassezia globosa; Malassezia obtusa;
Malassezia pachydermatis; Malassezia restricta; Malassezia
slooffiae; Malassezia sympodialis; Metarrhizium anisopliae;
Microsporum audouinii; Microsporum canis; Microsporum fulvum;
Microsporum gypseum; Microsporum persicolor; Mucor circinelloides;
Mucor hiemalis; Mucor racemosus; Mucor rouxii; Mucor spp;
Nattrassia mangiferae; Nectria haematococca; Onychocola canadensis;
Paecilomyces lilacinus; Paecilomyces spp; Paecilomyces variotii;
Paracoccidioides brasiliensis; Penicillium marneffei; Penicillium
spp; Phialophora spp; Phialophora verrucosa; Phoma spp; Pichia
anomala; Pichia etchellsii; Pichia guilliermondii; Pichia ohmeri;
Pithomyces spp; Pneumocystis carinii; Pseudallescheria boydii;
Ramichloridium obovoideum; Rhizomucor miehei; Rhizomucor pusillus;
Rhizomucor spp; Rhizopus arrhizus; Rhizopus microsporus; Rhizopus
oryzae; Rhizopus schipperae; Rhizopus spp; Rhodotorula
mucilaginosa; Rhodotorula rubra; Rhodotorula spp; Saccharomyces
cerevisiae; Saccharomyces spp; Sagrahamala spp; Saksenaea
vasiformis; Scedosporium apiospermum; Scedosporium prolificans;
Schizophyllum commune; Schizosaccharomyces pombe; Scopulariopsis
brevicaulis; Scytalidium dimidiatum Ulocladium spp; Sporobolomyces
spp; Sporothrix schenckii; Trichoderma spp; Trichophyton krajdenii;
Trichophyton mentagrophytes; Trichophyton raubitschekii;
Trichophyton rubrum; Trichophyton soudanense; Trichophyton spp;
Trichophyton terrestre; Trichophyton tonsurans; Trichophyton
verrucosum; Trichophyton violaceum; Trichosporon asahii;
Trichosporon beigelii; Trichosporon capitatum; Trichosporon
cutaneum; Trichosporon inkin; Trichosporon mucoides; Trichosporon
spp; Tritirachium spp; Wangiella dermatitidis or Yarrowia
lipolytica.
[0017] A substance "inhibits" growth of a cell (e.g., a fungal
cell) if it reduces the ability of the cell to grow, to any degree
(e.g., 10%, 25%, 50%, 75%, 90%, 99%, 100%), as compared to a
similar cell's growth in the absence of the substance.
[0018] A promoter's expression level is "modulated" if it is
increased or decreased to any degree.
Molecular Biology
[0019] In accordance with the present invention there may be
employed conventional molecular biology, microbiology, and
recombinant DNA techniques within the skill of the art. Such
techniques are explained in the literature. See, e.g., Sambrook,
Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual,
Second Edition (1989) Cold Spring Harbor Laboratory Press, Cold
Spring Harbor, N.Y. (herein "Sambrook, et al., 1989"); DNA Cloning:
A Practical Approach, Volumes I and II (D. N. Glover ed. 1985);
Oligonucleotide Synthesis (M. J. Gait ed. 1984); Nucleic Acid
Hybridization (B. D. Hames & S. J. Higgins eds. (1985));
Transcription And Translation (B. D. Hames & S. J. Higgins,
eds. (1984)); Animal Cell Culture (R. I. Freshney, ed. (1986));
Immobilized Cells And Enzymes (IRL Press, (1986)); B. Perbal, A
Practical Guide To Molecular Cloning (1984); F. M. Ausubel, et al.
(eds.), Current Protocols in Molecular Biology, John Wiley &
Sons, Inc. (1994).
[0020] A "polynucleotide", "nucleic acid" or "nucleic acid
molecule" includes the polymeric form of ribonucleosides
(adenosine, guanosine, uridine or cytidine; "RNA molecules") or
deoxyribonucleosides (deoxyadenosine, deoxyguanosine,
deoxythymidine, or deoxycytidine; "DNA molecules"), or any
phosphoester analogs thereof, such as phosphorothioates and
thioesters, in single stranded form, double-stranded form or
otherwise.
[0021] A "polynucleotide sequence", "nucleic acid sequence" or
"nucleotide sequence" is a series of nucleotide bases (also called
"nucleotides") in a nucleic acid, such as DNA or RNA, and means any
chain of two or more nucleotides.
[0022] A "coding sequence" or a sequence "encoding" an expression
product, such as a RNA, polypeptide, protein, or enzyme, is a
nucleotide sequence that, when expressed, results in production of
the product.
[0023] The term "gene" means a DNA sequence that codes for or
corresponds to a particular sequence of ribonucleotides or amino
acids which comprise all or part of one or more RNA molecules,
proteins or enzymes, and may or may not include regulatory DNA
sequences, such as promoter sequences, which determine, for
example, the conditions under which the gene is expressed. Genes
may be transcribed from DNA to RNA which may or may not be
translated into an amino acid sequence.
[0024] As used herein, the term "oligonucleotide" refers to a
nucleic acid, generally of no more than about 100 nucleotides
(e.g., 30, 40, 50, 60, 70, 80, or 90), that may be hybridizable to
a genomic DNA molecule, a cDNA molecule, or an mRNA molecule
encoding a gene, mRNA, cDNA, or other nucleic acid of interest.
Oligonucleotides can be labeled, e.g., by incorporation of
.sup.32P-nucleotides, .sup.3H-nucleotides, .sup.14C-nucleotides,
.sup.35S-nucleotides or nucleotides to which a label, such as
biotin, has been covalently conjugated. In one embodiment, a
labeled oligonucleotide can be used as a probe to detect the
presence of a nucleic acid. In another embodiment, oligonucleotides
(one or both of which may be labeled) can be used as PCR primers,
either for cloning full length or a fragment of the gene, or to
detect the presence of nucleic acids. Generally, oligonucleotides
are prepared synthetically, preferably on a nucleic acid
synthesizer.
[0025] A "protein sequence", "peptide sequence" or "polypeptide
sequence" or "amino acid sequence" refers to a series of two or
more amino acids in a protein, peptide or polypeptide.
[0026] "Protein", "peptide" or "polypeptide" includes a contiguous
string of two or more amino acids.
[0027] The term "isolated polynucleotide" or "isolated polypeptide"
includes a polynucleotide (e.g., RNA or DNA molecule, or a mixed
polymer) or a polypeptide, respectively, which is partially or
fully separated from other components that are normally found in
cells or in recombinant DNA expression systems or any other
contaminant. These components include, but are not limited to, cell
membranes, cell walls, ribosomes, polymerases, serum components and
extraneous genomic sequences.
[0028] An isolated polynucleotide or polypeptide will, preferably,
be an essentially homogeneous composition of molecules but may
contain some heterogeneity.
[0029] "Amplification" of DNA as used includes the use of
polymerase chain reaction (PCR) to increase the concentration of a
particular DNA sequence within a mixture of DNA sequences. For a
description of PCR see Saiki, et al., Science (1988) 239:487.
[0030] The term "host cell" includes any cell of any organism that
is selected, modified, transfected, transformed, grown, or used or
manipulated in any way, for the production of a substance by the
cell, for example the expression or replication, by the cell, of a
gene, a DNA or RNA sequence or a protein. For example, a host cell
may be a bacteria such as E. coli. The scope of the present
invention includes a host cell comprising any of the responsive
promoters or baseline promoters described herein along with fusions
comprising a responsive promoter or baseline promoter fused to a
reporter (e.g., a reporter gene). Preferably, the promoter or the
fusion is in a vector or is genetically integrated into a
chromosome of the host cell. A host cell can be, for example, a
Saccharomyces cerevisiae cell (e.g., strain MS1554 or strain
WT1554).
[0031] The sequence of a nucleic acid may be sequenced by any
method known in the art (e.g., chemical sequencing or enzymatic
sequencing). "Chemical sequencing" of DNA may denote methods such
as that of Maxam and Gilbert (Proc. Natl. Acad. Sci. USA (1977)
74:560), in which DNA is randomly cleaved using individual
base-specific reactions. "Enzymatic sequencing" of DNA may denote
methods such as that of Sanger (Sanger, et al., Proc. Natl. Acad.
Sci. USA (1977) 74:5463).
[0032] Aside from the responsive promoters of the invention,
nucleic acids of the invention can be flanked by other natural
regulatory (expression control) sequences, which may be associated
with heterologous sequences, including internal ribosome entry
sites (IRES) and other ribosome binding site sequences, enhancers,
response elements, suppressors, signal sequences, polyadenylation
sequences, introns, 5'- and 3'-non-coding regions, and the
like.
[0033] In general, a "promoter" or "promoter sequence" is a DNA
regulatory region capable of binding an RNA polymerase in a cell
(e.g., directly or through other promoter-bound proteins or
substances) and initiating transcription of a coding sequence. A
promoter sequence is, in general, bounded at its 3' terminus by the
transcription initiation site and extends upstream (5' direction)
to include the minimum number of bases or elements necessary to
initiate transcription at any level. Within the promoter sequence
may be found a transcription initiation site (conveniently defined,
for example, by mapping with nuclease S1), as well as protein
binding domains (consensus sequences) responsible for the binding
of RNA polymerase. The promoter may be operably associated with
other expression control sequences, including enhancer and
repressor sequences or with a nucleic acid of the invention.
[0034] A coding sequence (e.g., of a reporter) is "operably linked
to", "under the control of", "functionally associated with" or
"operably associated with" a transcriptional and translational
control sequence (e.g., a responsive promoter) in a cell when the
sequence directs RNA polymerase mediated transcription of the
coding sequence into RNA, preferably mRNA, which then may be RNA
spliced (if it contains introns) and, optionally, translated into a
protein encoded by the coding sequence.
[0035] The terms "express" and "expression" mean allowing or
causing the information in a gene, RNA or DNA sequence to become
manifest; for example, producing a protein by activating the
cellular functions involved in transcription and translation of a
corresponding gene. A DNA sequence is expressed in or by a cell to
form an "expression product" such as an RNA (e.g., mRNA) or a
protein. The expression product itself may also be said to be
"expressed" by the cell.
[0036] The term "transformation" means the introduction of a
nucleic acid into a cell. The introduced gene or sequence may be
called a "clone". A host cell that receives the introduced DNA or
RNA has been "transformed" and is a "transformant" or a "clone."
The DNA or RNA introduced to a host cell can come from any source,
including cells of the same genus or species as the host cell, or
from cells of a different genus or species.
[0037] The present invention includes vectors which comprise
polynucleotides of the invention (e.g., SEQ ID NOs: 1-41 or 52-68
or a functional variant thereof). The term "vector" includes a
vehicle (e.g., a plasmid) by which a DNA or RNA sequence can be
introduced into a host cell, so as to transform the host and,
optionally, promote expression and/or replication of the introduced
sequence. In general, a plasmid is circular, includes an origin
(e.g., 2 .mu.m origin) and, preferably includes a selectable
marker. In plasmids which can be maintained in yeast, the most
commonly used yeast markers include URA3, HIS3, LEU2, TRP1 and
LYS2, which complement specific auxotrophic mutations in a yeast
host cell, such as ura3-52, his3-D1, leu2-D1, trp1-D1 and lys2-201,
respectively. If the plasmid can be maintained in E. coli, it will
include a bacterial origin (ori) and a selectable market such as
the .beta.-lactamase gene (bla or AMP.sup.r). Commonly used
yeast/E. coli shuttle vectors are the Yip (see Myers et al., Gene
45: 299-310, (1986)), Yep (see Myers et al., Gene 45: 299-310,
(1986)), YCp and YRp plasmids. The YIp integrative vectors do not
replicate autonomously, but integrate into the genome at low
frequencies by homologous recombination. The YEp yeast episomal
plasmid vectors replicate autonomously because of the presence of a
segment of the yeast 2 .mu.m plasmid that serves as an origin of
replication (2 .mu.m ori). The 2 .mu.m ori is responsible for the
high copy-number and high frequency of transformation of YEp
vectors. The YCp yeast centromere plasmid vectors are autonomously
replicating vectors containing centromere sequences, CEN, and
autonomously replicating sequences, ARS. The YCp vectors are
typically present at very low copy numbers, from 1 to 3 per cell.
Autonomously replicating plasmids (YRp) which carry a yeast origin
of replication (ARS sequence; but not centromere) that allows the
transformed plasmids to be propagated several hundred-fold. YIp,
YEp, YCp and YRp are commonly known in the art and widely
available. Another acceptable yeast vector is a yeast artificial
chromosome (YAC). A yeast artificial chromosome is a biological
vector. It is an artificially constructed chromosome and contains
the telomeric, centromeric, and replication origin sequences needed
for replication in yeast cells (see Marchuk et al., Nucleic Acids
Res. 16(15):7743 (1988); Rech et al., Nucleic Acids Res. 18(5):1313
(1990)).
[0038] Vectors that could be used in this invention include
plasmids, viruses, bacteriophage, integratable DNA fragments, and
other vehicles that may facilitate introduction of the nucleic
acids into the genome of the host. Plasmids are the most commonly
used form of vector but all other forms of vectors which serve a
similar function and which are, or become, known in the art are
suitable for use herein. See, e.g., Pouwels, et al., Cloning
Vectors: A Laboratory Manual, 1985 and Supplements, Elsevier, N.Y.,
and Rodriguez et al. (eds.), Vectors: A Survey of Molecular Cloning
Vectors and Their Uses, 1988, Buttersworth, Boston, Mass.
[0039] A polynucleotides may be expressed in an expression system.
The term "expression system" means a host cell and compatible
vector which, under suitable conditions, can express a protein or
nucleic acid which is carried by the vector and introduced to the
host cell. Common expression systems include E. coli host cells and
plasmid vectors, insect host cells and Baculovirus vectors, and
mammalian host cells and vectors.
[0040] The present invention also contemplates any superficial or
slight modification to the responsive promoter and baseline
promoter nucleotide sequences of the invention. For example, the
present invention includes any "functional variant" of any
responsive promoter of the present invention (e.g., SEQ ID NOs:
1-41 or 68) or of any baseline promoter of the present invention
(e.g., SEQ ID NOs: 52-67). A functional variant of a promoter of
the present invention includes any sequence variant that retains
the ability to cause the expression of a downstream sequence (e.g.,
reporter such as SEQ ID NOs: 42-51) at any detectable level.
Methods for determining whether a particular promoter sequence
variant retains the ability to promoter expression of a down-stream
sequence are conventional and well known in the art.
[0041] The present invention contemplates sequence conservative
variants of the nucleic acids (e.g., SEQ ID NOs: 1-41 or 52-68).
"Sequence-conservative variants" of a polynucleotide sequence
(e.g., SEQ ID NOs: 1-41 or 52-68) are those in which a change of
one or more nucleotides results in no significant alteration in the
function of the promoter.
[0042] The present invention includes nucleic acids which hybridize
to the responsive promoter and baseline promoter polynucleotides of
the invention (e.g., SEQ ID NOs: 1-41 or 52-68). Preferably, the
nucleic acids hybridize under low stringency conditions, more
preferably under moderate stringency conditions and most preferably
under high stringency conditions. A nucleic acid molecule is
"hybridizable" to another nucleic acid molecule when a single
stranded form of the nucleic acid molecule can anneal to the other
nucleic acid molecule under the appropriate conditions of
temperature and solution ionic strength (see Sambrook, et al.,
supra). The conditions of temperature and ionic strength determine
the "stringency" of the hybridization. Typical low stringency
hybridization conditions may be 55.degree. C., 5.times.SSC, 0.1%
SDS, 0.25% milk, and no formamide; or 30% formamide, 5.times.SSC,
0.5% SDS. Typical, moderate stringency hybridization conditions are
similar to the low stringency conditions except the hybridization
is carried out in 40% formamide, with 5.times. or 6.times.SSC. High
stringency hybridization conditions are similar to low stringency
conditions except the hybridization conditions are carried out in
50% formamide, 5.times. or 6.times.SSC and, optionally, at a higher
temperature (e.g., 57.degree. C., 59.degree. C., 60.degree. C.,
62.degree. C., 63.degree. C., 65.degree. C. or 68.degree. C.). In
general, SSC is 0.15M NaCl and 0.015M Na-citrate. Hybridization
requires that the two nucleic acids contain complementary
sequences, although, depending on the stringency of the
hybridization, mismatches between bases are possible. The
appropriate stringency for hybridizing nucleic acids depends on the
length of the nucleic acids and the degree of complementation,
variables well known in the art. The greater the degree of
similarity or homology between two nucleotide sequences, the higher
the stringency under which the nucleic acids may hybridize. For
hybrids of greater than 100 nucleotides in length, equations for
calculating the melting temperature have been derived (see
Sambrook, et al., supra, 9.50-9.51). For hybridization with shorter
nucleic acids, i.e., oligonucleotides, the position of mismatches
becomes more important, and the length of the oligonucleotide
determines its specificity (see Sambrook, et al., supra,
11.7-11.8).
[0043] Also included in the present invention are polynucleotides
comprising nucleotide sequences which are at least about 70%
identical, preferably at least about 80% identical, more preferably
at least about 90% identical and most preferably at least about 95%
identical (e.g., 95%, 96%, 97%, 98%, 99%, 100%) to the reference
responsive promoter and baseline promoter polynucleotides (e.g.,
SEQ ID NO: 1-41 and 52-68) when the comparison is performed by a
BLAST algorithm wherein the parameters of the algorithm are
selected to give the largest match between the respective sequences
over the entire length of the respective reference sequences.
[0044] The following references regarding the BLAST algorithm are
herein incorporated by reference: BLAST ALGORITHMS: Altschul, S.
F., et al., J. Mol. Biol. (1990) 215:403-410; Gish, W., et al.,
Nature Genet. (1993) 3:266-272; Madden, T. L., et al., Meth.
Enzymol. (1996) 266:131-141; Altschul, S. F., et al., Nucleic Acids
Res. (1997) 25:3389-3402; Zhang, J., et al., Genome Res. (1997)
7:649-656; Wootton, J. C., et al., Comput. Chem. (1993) 17:149-163;
Hancock, J. M., et al., Comput. Appl. Biosci. (1994) 10:67-70;
ALIGNMENT SCORING SYSTEMS: Dayhoff, M. O., et al., "A model of
evolutionary change in proteins." in Atlas of Protein Sequence and
Structure, (1978) vol. 5, suppl. 3. M. O. Dayhoff (ed.), pp.
345-352, Natl. Biomed. Res. Found., Washington, D.C.; Schwartz, R.
M., et al., "Matrices for detecting distant relationships." in
Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3."
M. O. Dayhoff (ed.), pp. 353-358, Natl. Biomed. Res. Found.,
Washington, D.C.; Altschul, S.F., J. Mol. Biol. (1991) 219:555-565;
States, D. J., et al., Methods (1991) 3:66-70; Henikoff, S., et
al., Proc. Natl. Acad. Sci. USA (1992)89:10915-10919; Altschul, S.
F., et al., J. Mol. Evol. (1993) 36:290-300; ALIGNMENT STATISTICS:
Karlin, S., et al., Proc. Natl. Acad. Sci. USA (1990) 87:2264-2268;
Karlin, S., et al., Proc. Natl. Acad. Sci. USA (1993) 90:5873-5877;
Dembo, A., et al., Ann. Prob. (1994) 22:2022-2039; and Altschul, S.
F. "Evaluating the statistical significance of multiple distinct
local alignments." in Theoretical and Computational Methods in
Genome Research (S. Suhai, ed.), (1997) pp. 1-14, Plenum, N.Y.
Promoters and Screening Assays
[0045] The present invention provides several "gene responsive
promoters", "responsive promoters" or "biomarkers" which have been
identified in S. cerevisiae (e.g., SEQ ID NOs: 1-41 or 68). The
expression from each responsive promoter has been correlated with
the function of a particular gene which is essential in S.
cerevisiae. Below, in table 2, each gene ("gene" column) whose
temperature-sensitive allele was inactivated by shift to the
restrictive temperature is correlated with a promoter ("RP" column)
which was up-regulated in response to the shift along with the
function of the gene. The responsive promoters of the present
invention can be used to identify substances that modulate
(increase or decrease) the function of an essential fungal gene,
for example, in the assays described below. A substance which
modulates (increases or decreases) the function of any of the
essential genes mentioned herein is useful as an anti-fungal
compound which can be used, inter alia, to treat or prevent fungal
infections. For example, if the substance inhibits the activity of
the essential gene's translated product, it will lead to the death,
or decreased growth, of the host cell carrying the gene. Since
fungi, such as Saccharomyces cerevisiae, Aspergillus fumigatus,
Aspergillus niger, Aspergillus flavus and Candida albicans, are
genetically related, a substance which modulates an essential gene
in S. cerevisiae and kills the cell or inhibits its growth will be
lethal to or inhibit the growth of all of the cell types. A
substance which modulates the function of any of the essential
genes mentioned herein is also useful to the treatment or
prevention of cancer (e.g., by inhibition of the growth of a
malignant cell).
[0046] Substances which inhibit fungal cell growth or malignant
cell growth reduce it by any measurable degree. For example, a
substance which inhibits fungal cell or malignant cell growth can
kill the cell completely or reduce its level of growth by 10%. A
substance which inhibits fungal cell growth can be "fungistatic" in
that it inhibits growth of the cell to any measurable degree
without killing the cell.
[0047] One embodiment of the present invention includes a screening
assay wherein a biomarker of the present invention (e.g., SEQ ID
NO:1-41 or 68 or a functional variant thereof) is used to identify
a substance which inhibits fungal cell growth (a modulator of an
essential fungal gene) comprising the following steps: (a)
introducing a hybrid comprising a responsive promoter (e.g., SEQ ID
NO:1-41 or 68 or a functional variant thereof) operably linked to a
reporter (e.g., reporter gene such as SEQ ID NO: 42-51 or a
functional variant thereof) into a suitable cell (e.g., a fugal
cell); (b) contacting the cell with a substance to be tested for
the ability to inhibit fungal cell growth; (c) detecting expression
driven by the promoter in the hybrid; (d) selecting the substance
if it modulates expression from the responsive promoter.
[0048] If the level of expression driven by the promoter is
modulated (increased or decreased) in response to the substance,
this indicates that the substance does modulate the activity of the
essential gene and inhibits fungal cell growth.
[0049] In an embodiment of the invention, a negative-control
screening assay is also carried out. In this embodiment, a fungal
cell comprising the hybrid is contacted with the substance being
tested and another, separate cell comprising the same hybrid is
contacted with a blank (i.e., a substance known not to modulate the
expression level from the promoter) such as DMSO, water or buffer.
If the substance being tested increases or decreases the expression
level from the promoter in relation to the expression level from
the promoter/reporter in the cell contacted with the blank, this
indicates that the substance does modulate the expression level
from the responsive promoter, and, that it does modulate the
activity of the essential fungal gene (i.e., the substance is an
anti-fungal substance). A positive-control screening assay can also
be performed. In the positive-control assay, a fungal cell
comprising the hybrid is contacted with the substance being tested
and another, separate cell comprising the same hybrid is contacted
with a positive-control substance known to modulate the expression
level from the promoter. If the substance being tested and the
positive-control substance both increase or decrease the expression
level from the promoter, this indicates that the substance being
tested does modulate the expression level from the responsive
promoter, and, that it does modulate the activity of the essential
fungal gene (i.e., the substance inhibits fungal cell growth).
[0050] Another embodiment of the present invention includes a
ratiometric screening assay wherein a responsive promoter of the
present invention is used to identify a substance which inhibits
fungal cell growth (a modulator of an essential fungal gene)
comprising the following steps: (a) introducing, into a suitable
cell, a first hybrid comprising a first (responsive) promoter
selected from the group consisting of: S. cerevisiae YOR387C
Promoter, S. cerevisiae YMR175W Promoter, S. cerevisiae YFR026C
Promoter, S. cerevisiae YJL153C Promoter, S. cerevisiae YPL033C
Promoter, S. cerevisiae YOL058W Promoter, S. cerevisiae YOR255W
Promoter, S. cerevisiae YDR250C Promoter, S. cerevisiae YDR446W
Promoter, S. cerevisiae YDR536W Promoter, S. cerevisiae YOR255W
Promoter, S. cerevisiae YDL243C Promoter S. cerevisiae YDR256C
Promoter, S. cerevisiae YFL020C Promoter, S. cerevisiae YPL205C
Promoter, S. cerevisiae YGL205W Promoter, S. cerevisiae YGL117W
Promoter, S. cerevisiae YHR029C Promoter, S. cerevisiae YML116W
Promoter S. cerevisiae YJR109C Promoter, S. cerevisiae YCL030C
Promoter, S. cerevisiae YLR338W Promoter, S. cerevisiae YIL066W-A
Promoter, S. cerevisiae YNL093W Promoter, S. cerevisiae YLR381W
Promoter, S. cerevisiae YKL159C Promoter, S. cerevisiae YIL058W
Promoter, S. cerevisiae YKR037C Promoter, S. cerevisiae YNL279W
Promoter, S. cerevisiae YOR032C Promoter, S. cerevisiae YML058W
Promoter, S. cerevisiae YMR303C Promoter, S. cerevisiae YJL153C
Promoter S. cerevisiae YLR092W Promoter, S. cerevisiae YFL052W
Promoter, S. cerevisiae YIR017C Promoter, S. cerevisiae YLL062C
Promoter, S. cerevisiae YMR323W Promoter S. cerevisiae YPL171C
Promoter, S. cerevisiae YER065C Promoter and S. cerevisiae YDR114C
Promoter (e.g., SEQ ID NO: 1-41 or 68 or a functional variant
thereof); operably linked to a first reporter (e.g., SEQ ID NO:
42-51 or a functional variant thereof); and a second hybrid
comprising a second (baseline) promoter selected from the group
consisting of: S. cerevisiae YHR082C Promoter, S. cerevisiae
YDL126C Promoter S. cerevisiae YBR106W Promoter, S. cerevisiae
YLL043W Promoter, S. cerevisiae YPR108W Promoter, S. cerevisiae
YIL041W Promoter, S. cerevisiae YJL166W Promoter S. cerevisiae
YDR165W Promoter, S. cerevisiae YLR330 W Promoter, S. cerevisiae
YDL058W Promoter, S. cerevisiae YHR012W Promoter, S. cerevisiae
YKL053W Promoter, S. cerevisiae YKL196C Promoter, S. cerevisiae
YDL103C Promoter S. cerevisiae YLR427W Promoter and S. cerevisiae
YLL020C Promoter (e.g., SEQ ID NO: 52-67 or a functional variant
thereof); operably linked to a second reporter (e.g., SEQ ID NO:
42-51 or a functional variant thereof); (b) contacting the cell
with a substance to be tested for the ability to inhibit fungal
cell growth; (c) detecting expression driven by the first and
second promoters; and (d) selecting the substance if it causes the
first (gene responsive) promoter expression level to increase or
decrease in relation to the expression level from the second
(baseline) promoter.
[0051] In the ratiometric assay, a responsive promoter expression
level that does not increase or decrease in relation to the
expression level from the baseline promoter, in response to
contacting the cell with the substance being tested for anti-fungal
activity, indicates that the substance does not modulate the
expression level from the responsive promoter, and, that it does
not modulate the activity of the essential fungal gene. A
responsive promoter expression level that increases or decreases in
relation to the expression level from the baseline promoter, in
response to contacting the cell with the substance being tested for
anti-fungal activity, indicates that the substance does modulate
the expression level from the responsive promoter, and, that it
does modulate the activity of the essential fungal gene (i.e., it
inhibits fungal growth).
[0052] In an embodiment of the invention, a negative-control
ratiometric screening assay is also carried out. In this
embodiment, a cell comprising the first and second hybrid is
contacted with the substance being tested and another, separate
cell comprising the first and second hybrid is contacted with a
blank (i.e., a substance known not to modulate the expression level
from the first or second promoters) such as DMSO, water or buffer.
If the substance being tested increases or decreases the expression
level from the first promoter in relation to the expression level
from the second promoter and if the blank fails to increase or
decrease the expression level from the first promoter in relation
to the second promoter expression level, this indicates that the
substance does modulate the expression level from the responsive
promoter, and, that it does modulate the activity of the essential
fungal gene (i.e., the substance is an anti-fungal substance). A
positive-control ratiometic screening assay may also be performed.
In the positive-control assay, a cell comprising the first and
second hybrid is contacted with the substance being tested and
another, separate cell comprising the first and second hybrid is
contacted with a positive-control substance known to modulate the
expression level from the first promoter in relation to the
expression level from the second promoter. If the substance being
tested and the positive-control substance both modulate the
expression level from the first promoter in relation to the
expression level from the second promoter, this indicates that the
substance being tested does modulate the expression level from the
responsive promoter/reporter, and, that it does modulate the
activity of the essential fungal gene (i.e., the substance inhibits
fungal cell growth).
[0053] Up-regulation or down regulation of a given responsive
promoter, and identification of a substance which inhibits fungal
cell growth, can be assayed using an array. For example, a cell
comprising a responsive promoter/reporter hybrid or baseline
promoter/reporter hybrid can be contacted with a substance to be
tested for the ability inhibit fungal cell growth (to modulate an
essential fungal gene). Promoter expression can then be assayed
using conventional microarray assay techniques.
[0054] In one embodiment of the invention, a substance is
determined to inhibit fungal cell growth (modulate the activity of
an essential fungal gene) in a method as follows: (1) contacting a
suitable fungal cell (e.g., an S. cerevisiae cell) with a substance
to be tested for the ability to inhibit fungal cell growth
(modulate an essential fungal gene) wherein said cell comprises one
or more biomarkers operably linked to one or more reporter (e.g.,
the open reading frame which is located downstream of each
biomarker promoter in a wild-type cell); (2) obtaining mRNA from
the cell; (3) preparing labeled polynucleotides from the mRNA
(e.g., comprising a radiolabel (e.g., .sup.32P, .sup.35S, .sup.14C,
.sup.3H) or another suitable detectable label); (4) contacting the
labeled polynucleotide with an array, which comprises one or more
polynucleotides encoding said reporters (or oligonucleotide
fragments thereof), under conditions sufficient for labeled mRNA to
bind with corresponding polynucleotide probes on the array; (5)
optionally removing unbound, labeled mRNA from the array; and (6)
detecting bound, labeled mRNA.
[0055] The polynucleotide, in the array, can comprise a fragment of
the reporter polynucleotide, for example 10, 20, 30 or more
nucleotides thereof.
[0056] A "nucleic acid array", "array" or "matrix" refers to an
array, including a microarray (see infra), containing nucleic acid
probes, such as oligonucleotides or larger portions of genes. The
nucleic acid on the array is preferably single stranded. Arrays
wherein the probes are oligonucleotides are referred to as
"oligonucleotide arrays" or "oligonucleotide chips." A
"microarray", "biochip" or "biological chip" is an array of regions
having a density of discrete regions of at least about
100/cm.sup.2, and preferably at least about 1000/cm.sup.2. The
regions in a microarray have typical dimensions, e.g., diameters,
in the range of between about 10-250 .mu.m, and are separated from
other regions in the array by about the same distance.
[0057] The scope of the present invention includes an array (e.g.,
a microarray; discussed above) comprising one or more of said
reporters (discussed above) which are operably linked to one or
more of said biomarkers (discussed above).
[0058] Baseline promoters of the invention (e.g., SEQ ID NOs: 52-67
or a functional variant thereof) can also be used in assays for
determining if a substance inhibits fungal cell growth and to
calculate the inhibitory concentration (IC; e.g., IC50 or IC90) of
a particular substance. Since baseline promoters generally exhibit
a constant level of expression, they provide a convenient marker
for overall cellular growth or cellular mass. Generally, baseline
promoters can be used to determine the IC (e.g., IC50 or IC90) of a
given substance in a method including the following steps: (a)
introducing a hybrid comprising a baseline promoter selected from
the group consisting of: S. cerevisiae YHR082C Promoter, S.
cerevisiae YDL126C Promoter S. cerevisiae YBR106W Promoter, S.
cerevisiae YLL043 W Promoter, S. cerevisiae YPR108W Promoter, S.
cerevisiae YIL041W Promoter, S. cerevisiae YJL166W Promoter S.
cerevisiae YDR165W Promoter, S. cerevisiae YLR330 W Promoter, S.
cerevisiae YDL058W Promoter, S. cerevisiae YHR012W Promoter, S.
cerevisiae YKL053W Promoter, S. cerevisiae YKL196C Promoter, S.
cerevisiae YDL103C Promoter S. cerevisiae YLR427W Promoter and S.
cerevisiae YLL020C Promoter (e.g., SEQ ID NO: 52-67 or a functional
variant thereof); operably linked to a reporter (e.g., SEQ ID NO:
42-51 or a functional variant thereof) into a suitable fungal cell;
(b) contacting the cell with a substance to be tested for the
ability to inhibit growth of the cell; (c) detecting signal from
the reporter; and (d) selecting the substance if it causes a
reduction in the signal from the reporter and/or calculating the IC
(e.g., IC50 or IC90).
[0059] Optionally, this method can be performed with two or more
different baseline promoter/reporter fusions (e.g., in a
ratiometric assay discussed below). The IC (e.g., IC50 or IC90) can
then be determined two or more times when each separate reporter is
measured.
[0060] In an embodiment of the invention, a negative-control
screening assay is also carried out. In this embodiment, a cell
comprising the hybrid is contacted with the substance being tested
and another, separate cell comprising the same hybrid is contacted
with a blank (i.e., a substance known not to modulate the
expression level from the first or second promoters) such as DMSO,
water or buffer. If the substance being tested decreases the
reporter signal in relation to the signal from the reporter in the
cell contacted with the blank, this indicates that the substance is
an anti-fungal substance. A positive-control screening assay may
also be performed. In the positive-control assay, a cell comprising
the hybrid is contacted with the substance being tested and
another, separate cell comprising the same hybrid is contacted with
a positive-control substance known to modulate the reporter signal
(e.g., a known anti-fungal substance such as fluconazole,
itraconazole, moconazole or terbinafine). If the substance being
tested and the positive-control substance both decrease the
reporter signal, this indicates that the substance being tested is
an anti-fungal substance.
[0061] Baseline promoters of the invention (e.g., SEQ ID NOs: 52-67
or a functional variant thereof) can also be used in ratiometric
assays, comprising two or more baseline promoters operably linked
to two or more different reporters, for determining if a substance
inhibits fungal cell growth and to calculate the inhibitory
concentration (IC; e.g., IC50 or IC90) of a particular substance.
For example, a ratiometric assay comprising use of a baseline
promoter will comprise the steps of: (a) introducing two or more
hybrids comprising two or more different baseline promoters
selected from the group consisting of: S. cerevisiae YHR082C
Promoter, S. cerevisiae YDL126C Promoter S. cerevisiae YBR106W
Promoter, S. cerevisiae YLL043W Promoter, S. cerevisiae YPR108W
Promoter, S. cerevisiae YIL041W Promoter, S. cerevisiae YJL166W
Promoter S. cerevisiae YDR165W Promoter, S. cerevisiae YLR330 W
Promoter, S. cerevisiae YDL058W Promoter, S. cerevisiae YHR012W
Promoter, S. cerevisiae YKL053W Promoter, S. cerevisiae YKL196C
Promoter, S. cerevisiae YDL103C Promoter S. cerevisiae YLR427W
Promoter and S. cerevisiae YLL020C Promoter (e.g., SEQ ID NO: 52-67
or a functional variant thereof); operably linked to two or more
different reporters (e.g., SEQ ID NO: 42-51 or a functional variant
thereof) into a suitable fungal cell; (b) contacting the cell with
a substance to be tested for the ability to inhibit growth of the
cell; (c) detecting signal from the reporter; and (d) selecting the
substance if it causes a reduction in the signal from the reporter
and/or calculating the IC (e.g., IC50 or IC90). A range of several
concentrations of a given substance can be assayed using this
ratiometric method.
[0062] Ideally, the extent to which the reporter signal decreases,
in response to any given concentration of a substance which is
found to inhibit fungal cell growth, will be proportional (e.g., as
compared to the promoter's expression level in the absence of any
substance) to the extent to which the other reporter signals
decrease in response to the same concentration of substance.
[0063] In an embodiment of the invention, a negative-control
ratiometric screening assay is also carried out. In this
embodiment, a cell comprising the first and second hybrid is
contacted with the substance being tested and another, separate
cell comprising the first and second hybrid is contacted with a
blank (i.e., a substance known not to modulate the expression level
from the first or second promoters) such as DMSO, water or buffer.
If the substance being tested decreases the signal from the first
and second reporters and if the blank fails to decrease the signal
from the first and second reporters, this indicates that the
substance is an anti-fungal substance. A positive-control
ratiometic screening assay may also be performed. In the
positive-control assay, a cell comprising the first and second
hybrid is contacted with the substance being tested and another,
separate cell comprising the first and second hybrid is contacted
with a positive-control substance known to inhibit fungal growth
(e.g., a known anti-fungal substance such as fluconazole,
itraconazole, moconazole or terbinafine). If the substance being
tested and the positive-control substance both decrease the signal
from the first and second reporters, this indicates that the
substance being tested is an anti-fungal substance.
[0064] Using two or more baseline promoters is advantageous because
it facilitates elimination of false positives. For example, a false
positive signal, suggesting that a substance being tested inhibits
fungal cell growth, would occur if the substance causes a reduction
in expression from the baseline promoter without inhibiting fungal
cell growth. It is unlikely that the substance would have the
ability to reduce baseline promoter expression, without reducing
fungal cell growth (i.e., a false positive signal), with respect to
two different baseline promoters.
[0065] The term "inhibitory concentration" or "IC" refers to the
minimum concentration at which the substance being tested can
inhibit the growth of the organism being tested. The term includes
all inhibitory concentrations, for example, IC.sub.90 which is the
inhibitor concentration needed to inhibit growth of 90% of the
cells being tested and IC.sub.50 which is the inhibitor
concentration needed to inhibit growth of 50% of the cells being
tested. In an embodiment of the invention, a substance being tested
for anti-fungal or anti-malignant cell growth activity is selected
if it has a relatively low IC50 or IC90.
[0066] The present invention also includes compositions comprising
any one of the isolated responsive promoters (e.g., SEQ ID NOs:
1-41 or 68 or a functional variant thereof) or baseline promoters
identified herein (e.g., SEQ ID NOs: 52-67 or a functional variant
thereof). Also included in the present invention is any of the
isolated responsive promoters or baseline promoters identified
herein fused to a reporter (e.g., a reporter gene including, but
not limited to SEQ ID NOs: 42-51 or a functional variant
thereof).
[0067] A responsive promoter/reporter gene hybrid and/or baseline
promoter/reporter gene hybrid can be sustained in a host cell, in
which a screening assay of the invention is performed, by any
acceptable method known in the art. For example, the hybrid can be
genetically integrated into the chromosome of the host cell or it
can be maintained episomally, for example, in a plasmid (discussed
above) or yeast artificial chromosome (YAC; discussed above) or
other vector. In S. cerevisiae cells, for example, a hybrid can
easily be integrated into the HO locus. Methods for constructing
plasmids and vectors and for integrating a polynucleotide into a
host cell chromosome are conventional and very well known in the
art.
[0068] In another embodiment of the invention, anti-cancer agents
can be identified using a responsive promoter of the invention. In
particular, anti-cancer agents for treatment or prevention of
cancer (e.g., by inhibiting growth of a malignant cell) in a
mammal, preferably a human can be identified using a responsive
promoter of the invention. For example, the S. cerevisiae YLR338W
(SEQ ID NO: 22), S. cerevisiae YIL066W (SEQ ID NO: 23), S.
cerevisiae YNL093W (SEQ ID NO: 24), S. cerevisiae YDR250C (SEQ ID
NO: 8), S. cerevisiae YLR381W (SEQ ID NO: 25), S. cerevisiae
YBL065W (SEQ ID NO: 68), S. cerevisiae YMR323W (SEQ ID NO: 38) and
S. cerevisiae YPL171C (SEQ ID NO: 39) responsive promoters are
particularly useful for identification of anti-cancer agents. These
promoters have been found to modulate when genes in S. cerevisiae
biochemical pathways and functions, that are homologous to human
biochemical pathways and functions found to modulate the
development of cancer (e.g., the mitotic cluster, including
nucleocytoplasmic transport RAS, mitotic spindle, telomerase
maintenance, DNA damage and G1 cell cycle control, establishment of
cell polarity, exocyst complex and acetyltransferases). For
example, the scope of the invention includes methods for
identifying whether a substance can treat or prevent cancer in a
mammal (e.g., by inhibiting growth of a malignant cell) comprising
the following steps: (a) introducing a hybrid comprising a
responsive promoter selected from the group consisting of S.
cerevisiae YLR338W (SEQ ID NO: 22), S. cerevisiae YIL066W (SEQ ID
NO: 23), S. cerevisiae YNL093W (SEQ ID NO: 24), S. cerevisiae
YDR250C (SEQ ID NO: 8), S. cerevisiae YLR381W (SEQ ID NO: 25), S.
cerevisiae YBL065W (SEQ ID NO: 68), S. cerevisiae YMR323W (SEQ ID
NO: 38) and S. cerevisiae YPL171C (SEQ ID NO: 39) operably linked
to a reporter (e.g., reporter gene such as SEQ ID NO: 42-51 or a
functional variant thereof) into a suitable cell (e.g., a fugal
cell); (b) contacting the cell with a substance to be tested for
the ability to inhibit or prevent cancer; (c) detecting expression
driven by the responsive promoter in the hybrid; (d) selecting the
substance if it modulates expression from the responsive
promoter.
[0069] The present invention also includes a ratiometric screening
assay wherein a responsive promoter of the present invention is
used to identify a substance that can treat or prevent cancer in a
mammal (e.g., by inhibiting growth of a malignant cell) comprising
the following steps: (a) introducing, into a suitable fungal cell,
a first hybrid comprising a first (gene responsive) promoter
selected from the group consisting of: S. cerevisiae YLR338W (SEQ
ID NO: 22), S. cerevisiae YIL066W (SEQ ID NO: 23), S. cerevisiae
YNL093W (SEQ ID NO: 24), S. cerevisiae YDR250C (SEQ ID NO: 8), S.
cerevisiae YLR381W (SEQ ID NO: 25), S. cerevisiae YBL065W (SEQ ID
NO: 68), S. cerevisiae YMR323W (SEQ ID NO: 38) and S. cerevisiae
YPL171C (SEQ ID NO: 39); operably linked to a first reporter (e.g.,
SEQ ID NO: 42-51 or a functional variant thereof); and a second
hybrid comprising a second (baseline promoter) selected from the
group consisting of: S. cerevisiae YHR082C Promoter, S. cerevisiae
YDL126C Promoter S. cerevisiae YBR106W Promoter, S. cerevisiae
YLL043W Promoter, S. cerevisiae YPR108W Promoter, S. cerevisiae
YIL041W Promoter, S. cerevisiae YJL166W Promoter S. cerevisiae
YDR165W Promoter, S. cerevisiae YLR330 W Promoter, S. cerevisiae
YDL058W Promoter, S. cerevisiae YHR012W Promoter, S. cerevisiae
YKL053W Promoter, S. cerevisiae YKL196C Promoter, S. cerevisiae
YDL103C Promoter S. cerevisiae YLR427W Promoter and S. cerevisiae
YLL020C Promoter (e.g., SEQ ID NO: 52-67 or a functional variant
thereof); operably linked to a second reporter (e.g., SEQ ID NO:
42-51 or a functional variant thereof); (b) contacting the cell
with a substance to be tested for the ability to treat or prevent
cancer; (c) detecting expression driven by the first and second
promoters; and (d) selecting the substance if it causes the
expression level from the first (gene responsive) promoter to
increase or decrease in relation to the expression level from the
second (baseline) promoter.
[0070] In an embodiment of the invention, a negative-control
ratiometric screening assay is also carried out. In this
embodiment, a cell comprising the first and second hybrid is
contacted with the substance being tested and another, separate
cell comprising the first and second hybrid is contacted with a
blank (i.e., a substance known not to modulate the expression level
from the first or second promoters) such as DMSO, water or buffer.
If the substance being tested increases or decreases the expression
level from the first promoter/reporter in relation to the second
promoter/reporter and if the blank fails to increase or decrease
the expression level from the first promoter/reporter in relation
to the second promoter/reporter, this indicates that the substance
does modulate the expression level from the responsive promoter,
and, that it does modulate the activity of the fungal gene (i.e.,
the substance is an anti-cancer substance). A positive-control
ratiometic screening assay may also be performed. In the
positive-control assay, a cell comprising the first and second
hybrid is contacted with the substance being tested and another,
separate cell comprising the first and second hybrid is contacted
with a positive-control substance known to modulate the expression
level from the first promoter/reporter in relation to the
expression level from the second promoter/reporter. If the
substance being tested and the positive-control substance both
modulate the expression level from the first promoter/reporter in
relation to the expression level from the second promoter/reporter,
this indicates that the substance being tested does modulates the
expression level from the responsive promoter/reporter, and, that
it does modulate the activity of the essential fungal gene (i.e.,
the substance is an anti-cancer substance).
Reporters
[0071] One embodiment of the invention includes a responsive
promoter of the invention (e.g., SEQ ID NOs: 1-41 or 68 or a
functional variant thereof) or baseline promoter of the invention
(e.g., SEQ ID NOs: 52-67 or a functional variant thereof) fused to
a reporter. Any suitable reporter may be fused to the responsive
promoter or baseline promoter. A "reporter" is any polynucleic acid
which can be detected when expressed from a promoter. The term
includes reporter genes whose activity can be detected as well as
polynucleic acids which can be detected directly, for example, by
northern blot analysis. Alternatively, the translated protein
product of the expressed polynucleic acid can be detected by
western blot analysis. For example, the term "reporter" includes
the genetic sequence which is down-stream of a given responsive
promoter on a wild-type yeast chromosome. Reporters include
luminescent and fluorescent reporter genes including, but not
limited to, green fluorescent protein (e.g., SEQ ID NO: 48), lacZ
(e.g., SEQ ID NO: 51), Renilla luciferase (e.g., SEQ ID NO: 49; see
also Genbank Accession Nos.: AF416990; AR149562; AF362548 or
M63501) firefly (Photinus pyralis) luciferase (e.g., SEQ ID NO: 50;
see also Genbank Accession Nos.: U03687; M15077 and X84846), S.
cerevisiae ADE2 (e.g., SEQ ID NO: 42), S. cerevisiae LYS2 (e.g.,
SEQ ID NO: 43), S. cerevisiae TRP1 (e.g., SEQ ID NO: 44), S.
cerevisiae LEU2 (e.g., SEQ ID NO: 45), S. cerevisiae URA3 (e.g.,
SEQ ID NO: 46) and S. cerevisiae HIS3 (e.g., SEQ ID NO: 47).
[0072] The term "signal", in relation to a reporter, refers to the
indicia of expression of the reporter or the presence of the
reporter's gene product (e.g., protein or mRNA) in a sample. For
example, fluorescence is a signal that indicates the presence of
green fluorescent protein (GFP) and a "band" on photographic film
generated during a northern blot procedure is a signal indicating
the presence of a gene's RNA transcript. In an embodiment of the
invention, the expression driven from a given promoter fused to a
given reporter is measured by determining the signal from the
reporter.
[0073] Firefly luciferase may also be altered as described in
Leskinen et al. (Yeast. 20(13):1109-1113 (2003)) wherein the
carboxy-terminal peroxisomal targeting signal, Ser-Lys-Leu (slk),
of the firefly luciferase gene was removed; this allowed luciferase
activity to be detected in whole S. cerevisiae cells.
[0074] Other versions of luciferase which may be fused to a
responsive promoter of the invention (e.g., SEQ ID NOs: 1-41, 52-68
or a functional variant thereof) include the Vibrio harveyi luxA
(Genbank Accession No. M10961) and Vibrio harveyi luxB (Genbank
Accession No. M10961.1) genes, together, unfused or fused (i.e.,
luxAB or luxBA), Vibrio harveyi luxC, Vibrio harveyi luxD, Vibrio
harveyi luxE, Vibrio harveyi luxCDABE, the Photorhabdus luminescens
LuxCDABE operon (Genbank Accession No. M62917), Photorhabdus
luminescens LuxA, Photorhabdus luminescens LuxB, Photorhabdus
luminescens LuxC, Photorhabdus luminescens LuxD, Photorhabdus
luminescens LuxE, optionally expressed in the presence of the
Vibrio fischeri flavin oxidoreductase gene (frp; see Gupta et al.,
FEMS Yeast Res. 4(3):305-13 (2003)); Vibrio fischeri luxAB (see,
e.g., Yang et al., FEMS Microb. Lett. 176(1): 57-65 (1999)), the
Vibrio fischeri LuxCDABE operon (see e.g., Van Dyk et al., Appl.
Environ. Microbiol. 60:1414-1420 (1994); Belkin et al., Appl.
Environ. Microbiol. 62:2252-2256 (1996); Belkin et al., Water Res.
31:3009-3016 (1997); Genbank Accession No. AF170104); Vibrio
fischeri luxA, Vibrio fischeri luxB, Vibrio fischeri luxC, Vibrio
fischeri luxD or Vibrio fischeri luxE. Furthermore, a responsive
promoter or baseline promoter of the invention (e.g., SEQ ID NOs:
1-41 or 68 or a functional variant thereof) can be fused to a green
fluorescent protein (GFP)-luxAB hybrid gene.
[0075] The Ca.sup.2+ dependent photoprotein Aequorin from Aequorea
victoria (Campbell, K., Chemilluminescence; Ellis Horwood:
Chichester, England (1988); Ramanathan et al., Anal. Chim. Acta
369: 181-188 (1998); Witkowski et al., Anal. Chem. 66: 1837-1840
(1994); Galvan et al., Anal. Chem. 68:3545-3550 (1996)) can also be
fused to a responsive promoter or baseline promoter of the
invention (e.g., SEQ ID NOs: 1-41, 52-68 or a functional variant
thereof).
[0076] The KanMX selectable marker (e.g., from Tn903) can also be
fused to a responsive promoter or baseline promoter of the
invention (e.g., SEQ ID NOs: 1-41, 52-68 or a functional variant
thereof). Expression of the KanMX marker, in a cell, confers
resistance to G418 (geniticin; Wach et al., Yeast 10:1793-1808
(1994)).
[0077] The pat1 (phosphinothricin N-acetyl-transferase) selectable
marker can also be fused to a responsive promoter or baseline
promoter of the invention (e.g., SEQ ID NOs: 1-41, 52-68 or a
functional variant thereof; Goldstein & McCusker, Yeast
15:1541-1553 (1999)). Expression of the pat1 marker, in a cell,
confers resistance to bialaphos.
[0078] The nat1 (nourseothricin N-acetyl-transferase) selectable
marker can also be fused to a responsive promoter or baseline
promoter of the invention (e.g., SEQ ID NOs: 1-41, 52-68 or a
functional variant thereof; Goldstein & McCusker, Yeast
15:1541-1553 (1999)). Expression of the nat1 marker, in a cell,
confers resistance to nourseothricin.
[0079] The hph (hygromycin B phosphotransferase) selectable marker
can also be fused to a responsive promoter or baseline promoter of
the invention (e.g., SEQ ID NOs: 1-41, 52-68 or a functional
variant thereof; Goldstein & McCusker, Yeast 15:1541-1553
(1999)). Expression of the nat1 marker, in a cell, confers
resistance to hygromycin B.
[0080] The Sh ble selectable marker can also be fused to a
responsive promoter or baseline promoter of the invention (e.g.,
SEQ ID NOs: 1-41, 52-68 or a functional variant thereof).
Expression of the Sh ble marker, in a cell, confers resistance to
Zeocin.TM. (Phleomycin D1; Johansson & Hahn-Hagerdal, Yeast 19:
225-231 (2002)).
[0081] Other reporter genes which may be fused to a responsive
promoter or baseline promoter of the invention (e.g., SEQ ID NOs:
1-41, 52-68 or a functional variant thereof) include
.beta.-lactamase, for example, as described in Cartwright et al.
(Yeast 10(4):497-508 (1994)) wherein mature .beta.-lactamase was
fused to two segments of the K1 preprotoxin.
[0082] Furthermore, the E. coli .beta.-glucuronidase gene (GUS) can
be fused to a responsive promoter or baseline promoter of the
invention (e.g., SEQ ID NOs: 1-41, 52-68 or a functional variant
thereof) and used as a reporter gene, for example, in S. cerevisiae
(Marathe et al., Gene. 154(1):105-107 (1995)).
[0083] Chloramphenicol acetyltransferase (CAT) can be fused to a
responsive promoter or baseline promoter of the invention (e.g.,
SEQ ID NOs: 1-41, 52-68 or a functional variant thereof). CAT
radioassays are described, for example, by Sleigh (Anal. Biochem.
156(1):251-256 (1986)) and a non-radioactive CAT assay is described
by Young et al. (Anal. Biochem. 197(2):401-407 (1991)).
[0084] Construction of responsive promoter/reporter fusions and
baseline promoter/reporter fusions can be accomplished using
conventional molecular biological techniques which are commonly
known in the art. For example, Myers et al. (Gene. 1986;
45(3):299-310) disclose methods for fusing various promoters for
driving the expression of the E. coli lacZ gene.
[0085] Expression of reporters, such as green fluorescent protein,
luciferase or .beta.-galactosidase (lacZ) or any other reporter
mentioned herein, can be easily determined using any of the
numerous assays which are conventional and very well known in the
art. For example, Billinton et al. (Biosens. Bioelectron. 13(7-8):
831-8) describe the development of a green fluorescent protein
reporter in yeast. Dixon et al. (J. Steroid Biochem. Mol. Biol.
62(2-3): 165-71) describe an assay for determination of lacZ
expression in yeast. El Barkani et al. (Gene 246(1-2): 151-155
(2000)) describe Candida glabrata shuttle vectors suitable for
translational fusions to lacZ and use of .beta.-galactosidase as a
reporter of gene expression." Greer et al. (Luminescence 17(1):
43-74 (2002)) reviews the use of luciferase in expression assays.
Leskinen et al. (Yeast 20(13): 1109-13 (2003)) describes a one-step
measurement of firefly luciferase activity in yeast. Marathe et al.
(Gene 154(1): 105-7 (1995)) and Gallagher et al. ("Quantiation of
GUS activity by fluorometry" In: GUS Protocols: Using GUS gene as a
reporter of gene expression. Academic Press, San Diego, Calif.
(1992), pp. 47-59) discloses methods for assaying GUS reporter gene
expression. Pignatelli et al. (Biotechnol. Appl. Biochem. 27(Pt2):
81-88 (1998)) describe the expression and secretion of
.beta.-galactosidase in Saccharomyces cerevisiae using the signal
sequences of GgpI. Srikantha et al. (J Bacteriol 178(1): 121-9
(1996)) describes the use of Renilla reniformis luciferase as a
reporter gene in Candida albicans. Vanoni et al. (Biochem Biophys
Res Commun 164(3): 1331-8 (1989)) describes the use of E. coli
.beta.-galactosidase in Saccharomyces cerevisiae using the signal
sequence from the glucoamylase-encoding STA2 gene.
[0086] Alternatively, there are several auxotrophic reporters which
may be fused to a responsive promoter or baseline promoter of the
invention (e.g., SEQ ID NOs: 1-41, 52-68 or a functional variant
thereof). For example, ADE2, URA3, HIS3, LYS2, TRP1 or LEU2 can be
fused to the responsive promoter and introduced into a cell lacking
an endogenous copy of the gene (e.g., comprising a ura3-52,
his3-D1, leu2-D1, trp1-D1 or lys2-201 mutation). In general
activation of auxotrophic reporter genes can be determined, in a
cell lacking an endogenous copy of the auxotrophic marker, by
growing the cell on media lacking the nutrient whose cellular
synthesis is dependent on the auxotrophic reporter gene. In this
case, lack of expression of the auxotrophic marker (e.g., because
of lack of activation of the responsive promoter-auxotrophic marker
hybrid) will result in no growth; expression of the auxotrophic
marker (e.g., because of activation of the responsive
promoter-auxotrophic marker hybrid) will result in growth. For
example, activation of the markers ADE2, URA3, HIS3, LYS2, TRP1 or
LEU2 can be assayed on media lacking adenine, uracil, histidine,
lysine, tryptophan or leucine, respectively. In another embodiment,
expression of ADE2 can be conveniently determined by observing the
color of S. cerevisiae colonies containing the responsive
promoter-ADE2 hybrid. Specifically, ade2.sup.- S. cerevisiae cells
and colonies exhibit a red color phenotype. Expression of ADE2
(e.g., because of activation of the responsive promoter-ADE2
hybrid), in such a cell, will result in the expression of a white
color phenotype.
[0087] In an embodiment of the present invention, substances which
lead to a decrease in the expression level from a responsive
promoter of the invention can be negatively selected for. For
example, cell grown in the present of substance that decreases the
expression from a responsive promoter/URA3 hybrid can be selected
for by growth in the presence of 5-fluoroorotic acid (5-FOA). In
other embodiments of the invention, cells in which responsive
promoter/HIS3 hybrid expression is decreased, in response to a
substance, can be selected for by growing the cells in the presence
of 3-azatriazole. In yet another embodiment of the invention, cells
in which responsive promoter/LYS2 hybrid expression is decreased,
in response to a substance, can be selected for by growing the
cells in the presence of DL-alpha-aminoadipic acid. In a further
embodiment of the invention, cells in which responsive
promoter/CYH2 hybrid expression is decreased, in response to a
substance, can be selected for by growing the cells in the presence
of cycloheximide. In another embodiment of the invention, cells in
which responsive promoter/CAN/hybrid expression is decreased, in
response to a substance, can be selected for by growing the cells
in the presence of canavinine.
Transformation and Growth of Fungal Cells
[0088] Heterologous nucleic acids can be introduced into host yeast
cells by any of a number of known methods. These methods include,
but are not limited to, spheroplast transformation (Beggs, Nature
275:104 (1975); Cregg et al., Mol. Cell. Biol. 5:3376 (1985)),
lithium transformation (Hiep et al., Yeast 9:1189-1197 (1993);
Bogdanova et al., Yeast 11:343 (1995)), polyethylene glycol
transformation (Kleve et al., Gene 25:333-341 (1983); Dohmen et
al., Yeast 7:691-692 (1991); Dohmen et al., Yeast 7:691-692
(1991)), or electroporation.
[0089] Yeast can be grown on rich media or synthetic dropout media.
Drop-out media is synthetic media containing all essential
nutrients except for one or more. Drop-out media is commonly used
for selection of plasmids that contain genes for synthesis of these
components. For example, drop-out media lacking histidine can be
used to select for his3.sup.- host cells containing a plasmid
carrying a complementing HIS3 gene. A similar method can be
employed for selecting for fungal cells carrying a plasmid
containing the ADE2, URA3, LYS2, TRP1 or LEU2 gene, or any
combination thereof. For example, one liter of synthetic complete
media (with no nutrients dropped out) includes 20 g dextrose, 6.7 g
yeast nitrogen base without amino acids and 0.79 g Complete
Supplement Mixture CSM (Adenine 10 mg/liter; Arginine 50 mg/liter;
Aspartic Acid 80 mg/liter; Histidine 20 mg/liter; Isoleucine 50
mg/liter; Leucine 100 mg/liter; Lysine 50 mg/liter; Methionine 20
mg/liter; Phenylalanine 50 mg/liter; Threonine 100 mg/liter;
Tryptophan 50 mg/liter; Tyrosine 50 mg/liter; Uracil 20 mg/liter;
Valine 140 mg/liter; Qbiogene; Carlsbad, Calif.). A commonly used
"rich media" is YPD (Yeast-Peptone-Dextrose) or 1% yeast extract,
2% peptone, 2% dextrose. For solid media, 2% agar is included.
EXAMPLES
[0090] The following examples are provided to more clearly describe
the present invention and should not be construed to limit the
scope of the invention in any way. Any and all reagents (e.g.,
plasmids, primers, cells) described herein form part of the present
invention along with any methods described herein.
Example 1
Identification of Biomarkers
[0091] In this example, genes identified by "Affymetrix Microarray"
(Affymetrix, Inc.; Santa Clara, Calif.) technology are used as
biomarkers to identify compounds that affect a target protein. By
fusing a biomarker promoter to a reporter gene, assays can be
developed to screen for antifungal compounds. In addition, several
"baseline promoters" have been identified which exhibit a constant
level of expression when temperature is shifted.
[0092] Yeast temperature-sensitive shift. Saccharomyces cerevisiae
strains were grown at 30.degree. C. in YPD (1% yeast extract, 2%
peptone, 2% glucose). S. cerevisiae wild type WT1554 and ts-mutant
strains were streaked out from permanent stock and grown on YPD
plates (including 2% agar) at 30.degree. C. until colonies formed.
Several colonies were picked to start 10 ml cultures, grown up
overnight at 30.degree. C. and used to inoculate 25 mls of
30.degree. C. YPD to an OD of 0.15. Cultures were incubated at
30.degree. C. for 2 hrs and then heat shifted to 37.degree. C. by
the addition of 25 mls of 46.degree. C. YPD. Strains were grown for
2 hr, 4 hr, 6 hrs and cells were collected for determination of
Affymetrix Microarray expression profiles. Cells were harvested by
centrifugation for 3 minutes at 3500 rpm, resuspend in 6 ml of ice
cold ddH.sub.2O centrifuged for 3 minutes at 3500 rpm, the
supernatant discarded and the cell pellet flash frozen in a dry ice
ethanol bath. Pellets were stored at -80.degree. C. until total RNA
preparation.
[0093] Isolation of Saccharomyces cerevisiae total RNA. Total RNA
was isolated with slight modifications by the method of Schmitt et
al. (Nucleic Acids Res. 18(10):3091-3092 (1990)). Cell pellets were
removed from the -80.degree. C. freezer and 1.6 ml of pre-warmed
65.degree. C. acid phenol-chloroform-isoamylalcohol (50:50:1,
pH4.7; (Sigma P-1944) and 1.6 ml of TES (10 mM Tris pH 7.5, 10 mM
EDTA, 0.5% SDS) were added and the pellet was resuspended by
vortexing. Tubes were incubated at 65.degree. C. in a waterbath for
1 hour with vortexing for 20 seconds every 10 minutes. The mixture
was transferred to Phaselock tubes (Eppendorf) and centrifuged for
20 minutes at room temperature. Mixtures were re-extracted with 750
.mu.l of acid phenol-chloroform-isoamylalcohol per tube,
recentrifuged then extracted with 750 .mu.l 24:1 chloroform:Isoamyl
alcohol (Sigma C-0549) and centrifuged for 10 minutes room
temperature. The aqueous phase was transferred to an eppendorf tube
and total RNA was precipitated by the addition of 50 .mu.l of 3M
Sodium acetate (NaOAc) pH 5.2 and 1 ml of 100% EtOH followed by
incubation at -20.degree. C. for 1/2 hr. Total RNA pellets were
washed with 500 .mu.l of 80% ETOH and the pellet was resuspended in
50 .mu.l of 65.degree. C. RNAsecure (Ambion; Austin, Tex.) and
incubated for 10 min at 65.degree. C. to inactivate RNAses. Total
RNA quality and quantity was determined by measurement of the
OD.sub.260/OD.sub.280 absorbance ratio followed by analysis of RNA
utilizing Agilent RNA chips. RNA was stored at -80.degree. C. prior
to Affymetrix labeling and hybridization.
[0094] Affymetrix total RNA labeling and Affymetrix Yeast Genome
YG_S98 microarray probing. Polynucleotide Expression Profiling.
Total S. cerevisiae RNA from each RNA sample (see supra) was used
to prepare biotinylated target RNA according to the Affymetrix
Genechip.RTM. Expression Analysis Technical Manual, 2003.
http://www.affymetrix.com/support/technical/anual/manual/expression_manua-
l.affx). Briefly, 5 .mu.g of total RNA was used to generate
first-strand cDNA by using a T7-linked oligo(dT) primer. After
second-strand synthesis probes were generated by in vitro
transcription with biotinylated UTP and CTP (Enzo Diagnostics).
Biotinylated probes were hybridized to Affymetrix Yeast YG_S98 DNA
Microarrays (Affymetrix Inc., Santa Clara, Calif.). Target cDNA
generated from each sample were then processed according to the
manufacturer's recommended procedures using an Affymetrix GeneChip
Instrument System
(http://www.affymetrix.com/support/technical/manual/expression_manual.aff-
x). Briefly, spike controls were added to 5 .mu.g fragmented cRNA
before overnight hybridization to Affymetrix Yeast Genome YG_S98
microarrays. Microarrays were then washed and stained with
streptavidin-phycoerythrin, before being scanned on an Affymetrix
GeneChip scanner. The GeneChip.RTM. Yeast Genome YG_S98 Array
contains probe sets for approximately 6,400 S. cerevisiae (S288C
strain) genes identified in the Saccharomyces Genome Database. This
array also contains approximately 600 additional probe sets
representing putative open reading frames (ORFs) identified by SAGE
analysis, mitochondrial proteins, TY proteins, plasmids, and a
small number of ORFs for strains other than S288C (Affymetrix Santa
Clara, Calif.). Scanned image files were inspected for artifacts
and analyzed with GeneChip.RTM. Expression Analysis software
(Affymetrix, Santa Clara, Calif.). Each chip was scaled to 150
(see, Affymetrix, Genechip.RTM. Expression Analysis Technical
Manual, 2003) to account for minor differences in microarray
hybridization intensity, so that overall expression levels for each
yeast strain could be compared.
[0095] Identification of up-regulated gene biomarkers in heat
shifted S. cerevisiae ts-alleles by analysis of Affymetrix
microarrays by GeneSpring analysis software (Silicon Genetics).
Affymetrix Yeast Genome YG_S98 microarray data were loaded onto the
GeneSpring analysis software program 6.0 (Silicon Genetics, Redwood
City, Calif.). Microarray data from temperature shifted S.
cerevisiae ts-alleles from the 2, 4, and 6 hr time-points (see
supra) were normalized to S. cerevisiae wild type strain MS1554
from 2, 4, and 6 hr time-points. Up-regulated gene biomarkers were
identified in S. cerevisiae ts-alleles that showed a fold increase
in expression at a single time-point as compared to wild-type S.
cerevisiae strain MS1554. Data were further analyzed to identify
up-regulated gene biomarkers that showed up-regulation in
expression at 2, 4, and 6 hr time-points by graphical analysis of
identified genes. In addition genes that showed up-regulation in
expression were analyzed in all ts-alleles to identify genes that
were uniquely up-regulated in expression in a single ts-allele or
up-regulated in a group of ts-alleles.
Plasmids and Constructs
(a) Generation PHO5 Signal Sequence Tagged Renilla luciferase
[0096] pSPRT17 (Cyc1A term to HO-poly-HO vector). The cyclA
terminator (REF) was PCR amplified as a 371 by fragment from pYES2
(Invitrogen) using PCR primers PR131 (5'
tttgcgcgcGGGCCGCATCATGTAATTAGT 3') (SEQ ID NO: 69) and PR132 (5'
aaaagatctGACCGA GCGCAGCGAGTCAGT 3') (SEQ ID NO: 70) designed to
BssHII and BglII. and cloned into the TOPO-TA to yield pSPRT19. The
371 by fragment from pSPRT19 was ligated to BssHII and BglII.
digested HO-poly-HO (Voth et al., Nuc. Acid Res. 29(12): e59
(2001)) to yield pSPRT17.
[0097] pSPRT47 (SEQ ID NO: 85) (Renilla cloned into pSPRT17). The
renilla luciferase gene was amplified by PCR as a 1004 by fragment
amplified from phRG-B (Promega) using primers PR161 (5'
CCCATGTTAATTAAATCTGTTGTTTATTCAATTTTAGCCGCTTCTTTGGCCAATGCAGGTGCTTCCAAGGTGT-
ACGACCCCGA 3') (SEQ ID NO: 71) and PR162 (5'
AAAGGCGCGCCTTACTGCTCGTTCTTCAGCAC 3') (SEQ ID NO: 72), designed to
contain the PHO5 signal sequence (MLIKSVVYSILAASLANAG) (SEQ ID NO:
73) and PacI/AscI ends. cloned into the TOPO-TA to yield pSPRT27.
The PacI/AscI fragment from pSPRT27 was ligated to PacI/BssHII
digested pSPRT17 to yield pSPRT47. The Pad site was designed to
create an ATG start codon fusion with Pad adapted promoters. This
added LI to the signal sequence. The PHO5 tagged luciferase was
designated PHO5.hRluc.
[0098] pSPRT168 (Gall promoter PHO5.hRluc). The GAL1 promoter
region was PCR amplified as a BamHI/PacI 474 by fragment using PCR
primers PR172 (5'TTTGGATCCACGGATTAGAAGCCGCCGAGCG 3') (SEQ ID NO:
74) and PR 173B (5' CCCTTAATTAACATGGTTTTTTCTCCTTGACGTTA 3') (SEQ ID
NO: 75) cloned into the TOPO-TA to yield pSPRT33B. The BamHI/PacI
fragment was ligated to BamHI/PacI digested pSPRT47 to yield
pSPRT167. pSPRT167 was introduced into S. cerevisiae to yield
strain Y167.
[0099] pSPRT207 (YHR082C promoter PHO5.hRluc). The YHR082C promoter
was PCR amplified as a 1.1 kb BamHI/PacI fragment using PCR primers
PR207 (5' TTTGGATCCTGTGCACGATGGCTTCGTTTTAGCC 3') (SEQ ID NO: 76)
and PR208 (5' CTTAATTAACATCGAGTTTCAAAAAATTGTAAG 3') (SEQ ID NO: 77)
and cloned into the TOPO-TA to yield pTOPO207. The 1.1 kb
BamHI/PacI fragment from pSPRT207TOPO was ligated to BamHI/PacI
digested pSPRT47 to yield pSPRT207. pSPRT207 was introduced into S.
cerevisiae to yield strain Y207.
[0100] pSPRT211 (ERG10 promoter PHO5.hRluc). The ERG10 promoter was
PCR amplified as a 397 bp BamHI/PacI fragment using PCR primers
PR213 (5' CCCGGATCCTCTACGATATATCCTGTAAATAG 3') (SEQ ID NO: 78) and
PR214 (5' CTTAATTAACATTTTGAGTACGTCTAATCTGTAT 3') (SEQ ID NO: 79)
and cloned into the TOPO-TA to yield pTOPO211. The 397 by
BamHI/PacI fragment from pSPRT213TOPO was ligated to BamHI/PacI
digested pSPRT47 to yield pSPRT211. pSPRT211 was introduced into S.
cerevisiae to yield strain Y211.
(b) Generation PHO5 Signal Sequence Tagged FireFly Luciferase
[0101] pSPRT50 (SEQ ID NO: 86) (Firefly luciferase cloned into
pSPRT17). The firefly luciferase gene was PCR as a 1710 by fragment
amplified from pGL2-Basic (Promega; Madison, Wis.) using primers
PR167 (5' CCCATGGGATCCAAATCTGTTGTTTATTCAATTTTA
GCCGCTTCTTTGGCCAATGCAGGTGAAGACGCCAAAAACATAAAGAAA 3') (SEQ ID NO:
80) and PR168SLK (5'AAAGGCGCGCCTTACTTTCCGCCCTTCTTGGCCT 3') (SEQ ID
NO: 81), designed to contain the PHO5 signal sequence
(MLIKSVVYSILAASLANAG) (SEQ ID NO: 82) and BamHI/AscI ends and
remove the C-terminal SLK peroxisomal targeting sequence (Leskinen
et al., Yeast 20(13):1109-1113 (2003)). This fragment was cloned
into the TOPO-TA to yield pSPRT50. The BamHI/AscI fragment from
pSPRT50 was ligated to BamHI/BssHII digested pSPRT17 to yield
pSPRT50. The BamHI site was designed to create an ATG start codon
fusion with BamHI adapted promoters. The PHO5 tagged firefly
luciferase was designated PHO5.firefly.
[0102] pSPRT170 (Gall promoter PHO5.firefly). The Galp1 promoter
region was PCR amplified as a BamHI 472 by fragment using PCR
primers PR174 (5'TTTGGATCCACGGATTAGAAGCCGCCGAGCG 3') (SEQ ID NO:
83) and PR175B (5' TTTGGATCCCATGGTTTTTTCTCCTTGACGTTA 3') (SEQ ID
NO: 84) cloned into the TOPO-TA to yield pTOPO174. The BamHI
fragment was ligated to BamHI digested pSPRT50 to yield pSPRT170.
pSPRT170 was introduced into S. cerevisiae to yield strain
Y170.
TABLE-US-00002 TABLE 2 Biomarkers identified. Fold Essential 2, 4,
or Gene Function Affy ID 6 hr RP Synonym Function YCR052W Chromatin
8141_AT 15 YOR387C remodeling complex YBR156C Mitotic spindle
protein 8141_AT 5 YOR387C YDR166C Exocyst complex 9445_AT 13
YMR175W SIP18 Salt induced protein YDR166C Exocyst complex 5323_AT
YFR026C YDR166C Exocyst complex 11185_AT 17 YJL153C INO1
L-myo-inositol-1-phosphate synthase YKR025W RNA polymerase III
7810_AT 9.2 YPL033C subunit YKR025W RNA polymerase III 8593_AT 7.0
YOL058W ARG1 Arginosuccinate synthetase subunit YDL105W 8276_AT 25
YOR255W YDL105W 6215_AT 14 YDR250C YDL105W 6007_AT 9 YDR446W ECM11
ExtraCellular Mutant YDL105W 5915_AT 8 YDR536W STL1 sugar
transporter-like protein YDR016C spindle integrit, 8276_AT 16
YOR255W kinetochore YDR016C spindle integrit, 6710_AT 9 YDL243C
AAD4 Putative aryl-alcohol dehydrogenase kinetochore YDR016C
spindle integrit, 6221_AT 9 YDR256C CTA1 Catalase A kinetochore
YDR016C spindle integrit, 6215_AT 9 YDR250C kinetochore YDR016C
spindle integrit, 6007_AT 11 YDR446W ECM11 posible chromatin
structure maint kinetochore YDR016C spindle integrit, 5915_AT 10
YDR536W STL1 sugar transporter-like protein kinetochore YNL131W
Translocase 5410_F_AT 5.115 YFL020C PAU5 member of the seripauperin
Mitochondrial proteinVgene family (see Gene_class membrane PAU)
YDR464W Control expression of 7953_AT 5.8 YPL205C spliceosome
YPL128C TTAGGG repeat 5220_AT 13.5 YGL205W POX1 fatty-acyl coenzyme
A oxidase binding factor YLR291C Translational initiation 5125_AT
8.9 YGL117W factor YLR291C Translational initiation 4503_AT 8.4
YHR029C factor YLR291C Translational initiation 9771_AT 5.8 YML116W
ATR1 predicted protein is very hydrophobic, factor has many
membrane-spanning regions, several potential glycosylation sites,
potential ATP-binding site YLR291C Translational initiation
10902_AT 5.4 YJR109C CPA2 carbamyl phosphate synthetase factor
YLR291C Translational initiation 6902_AT 5.3 YCL030C HIS4
histidinol dehydrogenase factor YLR293C Nuclear organization
10008_AT 71.13 YLR338W YLR293C Nuclear organization 4039_AT 38.90
YIL066W-A RNR3 Ribonucleotide reductase YLR293C Nuclear
organization 8939_AT 32.54 YNL093W YPT53 rab5-like GTPase YLR293C
Nuclear organization 6215_AT 25.45 YDR250C YLR293C Nuclear
organization 9962_AT 12.40 YLR381W Kinetochore protein YLR293C
Nuclear organization 10714_AT 10.96 YKL159C Regulator of
calcineurnin YLR293C Nuclear organization 4159_AT 10.96 YIL058W
YLR293C Nuclear organization 10554_AT 9.85 YKR037C Spindle pole
component YDR499W Telomere Checkpoint 10008_AT 125.0 YLR338W
YDR499W Telomere Checkpoint 6215_AT 43.4 YDR250C YDR499W Telomere
Checkpoint 4039_AT 42.8 YIL066W-A RNR3 Ribonucleotide reductase
YDR499W Telomere Checkpoint 9160_AT 28.9 YNL279W
Pheromone-regulated multispanning YDR499W Telomere Checkpoint
8939_AT 22.2 YNL093W YPT53 rab5-like GTPase sorting and endocytosis
YDR499W Telomere Checkpoint 9962_AT 15.3 YLR381W Kinetochore
protein YDR499W Telomere Checkpoint 10554_AT 9.7 YKR037C Spindle
pole component YDR499W Telomere Checkpoint 8505_AT 9.6 YOR032C HMS1
myc-family transcription factor homolog YBR102C exocyst complex
10008_AT 109.7 YLR338W YBR102C exocyst complex 9160_AT 58.7 YNL279W
Pheromone-regulated multispanning YBR102C exocyst complex 6215_AT
53.7 YDR250C YBR102C exocyst complex 4039_AT 37.5 YIL066W-A RNR3
Ribonucleotide reductase YBR102C exocyst complex 8939_AT 27.6
YNL093W YPT53 rab5-like GTPase YGL225W May regulate Golgi 9293_AT
6.3 YML058W SML1 Suppressor of mec lethality function YGR065C
Vitamin H transporter 9309_AT 5.49 YMR303C ADH2 alcohol
dehydrogenase II YML091C Mitochondrial RNase P 11185_AT 23.0
YJL153C INO1 L-myo-inositol-1-phosphate synthase YML091C
Mitochondrial RNase P 10256_AT 16.2 YLR092W SUL2 high affinity
sulfate permease YML091C Mitochondrial RNase P 5428_AT 15.8 YFL052W
YML091C Mitochondrial RNase P 4098_AT 5.2 YIR017C MET28
Transcriptional activator of sulfur amino acid metabolism YML091C
Mitochondrial RNase P 10414_AT 4.2 YLL062C YDR081C Regulates trans
of 10008_AT 112 YLR338W PDC1, PDC5 YDR081C Regulates trans of
6215_AT 42 YDR250C PDC1, PDC5 YDR081C Regulates trans of 4039_AT 35
YIL066W RNR3 Ribonucleotide reductase subunit PDC1, PDC5 YDR081C
Regulates trans of 8939_AT 21 YNL093W YPT53 rab5-like GTPase PDC1,
PDC5 YMR033W Transcriptional 10008_AT 57 YLR338W regulation YMR033W
Transcriptional 6215_AT 34 YDR250C regulation YMR033W
Transcriptional 4039_AT 22 YIL066W-A RNR3 Ribonucleotide reductase
subunit regulation YMR033W Transcriptional 9288_S_AT 19.6 YMR323W
regulation YMR033W Transcriptional 7944_AT 12 YPL171C OYE3 DAD(P)H
dehydrogenase regulation YMR033W Transcriptional 5636_AT 9 YER065C
ICL1 Isocitrate lyase regulation YMR033W Transcriptional 6349_AT 9
YDR114C regulation
[0103] Responsive promoters identified in the experiments set forth
above are as follows:
TABLE-US-00003 >YOR387C Chr 15 reverse complement (SEQ ID NO: 1)
GCCAGGGAAGCCTATATCTGATTCCCTGTTTCATAATCCAATGCAG
CCACTAGCTTATAATTATTTGAACTATTTGTCGAACATCACAGTAA
TAAAATCCCCAGAAAGTTCCACTTGCTGCATATTGGCACCTGTTGA
TTCACTCTCCATCACTTTTTTGTTAGCCGCCCAGCCTAGAAAGTCT
TTAAATACATCTGAAATTTTTTTTTTTTTAACAGTGCACCCGTGCA
TCATACCTCATGCAAGGTACCTTTTTTTCTCAAAGGTATTGTCTTC
CATTGAAGTGGCACTATGGCATGATGAACCCTGAGCATTTCTGAAT
TCAACAGAACCAAATTGTCCAGAAATAAATCTGTCCGACATGAATT
ATGAAACTTTTTTTCAATTAAGTGAAGAGAATTTTGCAGCGTCTTA
CCATTATTTTGACCCATTGGTCGCATGTTTGCGCTTTGACTTCGAG
AACCATGTTAAAGCTTACTTGTACGACAACCAATGAAGTATATTAC
GGCAGTTTTTTTGGACTGGGTCAAAAAAAGTGTTGCATAATCAAAT
CAGGAACACATTAAAATGTTGTAAAATTTGTCTTAGTATCACCTGA
GTGGTTATTCATTACGTACTACTGTCAAAATATTCGGATCTTTCCT
AAACGGGCTTTTGAATTAGTGTTGCTTCTATTCCTGGAATGGAAGG
TAATACTTTCATGCATTCTCGCTTTTCGGACTTTTAACAATAAATT
AAAAACAATGATACTTTCATCAACTACCTATACACCCTGCGGGTCA
ATTATTTTTTTTTCGAATAACCGCTGAGGTTGGAAATATAGAAACA
TATTCCAGATCTGTATTTTCAGTTGCTAGAAAAAGGTTAATATAAT
CATTAAGGTTTTCAGCATATAACAGGTATAATTGATATATAAGCAT
CGTAATTTTCATTCAAAATGGAGAGCTACTGCTTCTGATAGATTGT
ACAATCTCAAGAAATCAAGAACAACAACCATACCATGAGT >YMR175W Chr 13 (SEQ ID
NO: 2) TTGTGTTTAGACTTAAGTATGAAAATTTTATGTATGAGCTGTGGCT
ATGTATCCGCTGGCAAATAGGTCTGCTTTTTCTACACTTTCCACCC
TCAACCTAACAGAGCCCGCTGGCACAAATAATCGATAGTAGGACAA
CAGAGCTACTCCTTCTTATGCCCCGCCCCTTTGAGCTTGTTGTATT
GCTCTTGATAGTTGTGTTTTTCACTTTCATCAGCATCGGTAGTCTT
GCCGTCCTTGTCTTGACTAGCCATTTTCTTAAAAGCGTCACTCACT
ACCTTTGCATCGCCCTGCAATTTTTCCTTTGAGCTCTGAAATATTT
CGCTCACTTTTTGTTGGTCTGTATTCATTCTGGATGTCTTGGTTGT
AGAAATTTCTTTTATTGGTTCATTAAAGTCAAGGTAAATGGCGAGA
ACTAGAATAGAGTTTTATTCTTTTTACCGTTATATAGATAATTCTA
GCCGGGGGCGGTCGCCCCTGAGATTCCCGACATCAGTAAGACATAG
TACTGTACGATTACTGTACGATTAATCTATCCACTTCAGATGTTCA
ACAATTCCTTTTGGCATTACGTATTAATACTTCATAGGATCGGCAC
CCTCCCTTAAGCCTCCCCTAAATGCTTTCGGTACCCCTTTAAGACA
ACTATCTCTTAACCTTCTGTATTTACTTGCATGTTACGTTGAGTCT
CATTGGAGGTTTGCATCATATGTTTAGGTTTTTTTGGAAACGTGGA
CGGCTCATAGTGATTGGTAAATGGGAGTTACGAATAAACGTATCTT
AAAGGGAGCGGTATGTAAAATGGATAGATGATCATGAATACAGTAC
GAGGTGTAAAGAATGATGGGACTGAGAGGGCAATTATCATCCCTCA
GAATCAACATCACAAACATATATAAAGCTCCCAATTCTGCCCCAAA
GTTTTGTCCCTAGGCATTTTTAATCTTTGTATCTGTGCTCTTTACT
TTAGTAGAAAGGTATATAAAAAAGTATAGTCAAGATG >YFRO26C Chr 6 reverse
complement (SEQ ID NO: 3)
GAGGTTTCAACATGCCAACAATGGAAGTGGTTAAGAGACGTTTTCT
GGTTTCTGGGCAGCTGTTACATAGAATCTCTGATTTACATGCTCAA
GTATTCGCAAAGATCGGTATGATGGGAAAATAGGTGAATTTTGGGA
TAATGACTGAGATTATACCGTTGTTAATGCTATAATATTAATTATA
CAGAATATATTAGAAGTTCTCCTCGAGGATATGGGAACCCACAGAA
GCGAATCGATGTTTCTACATAAAAGAAAAAAATAATCTCATTTTAT
CTTCGGTTTTGAATCTTTTCATTTTTTTTTCCTATTACATTATCAA
TTCTGGCATTTCAGTGTTTATAAAAAGATCAAATTGCTGTTTGAAA
CTCATTGCGCTAATTCCATAATTTTGCACCATGTATGCTCATGACT
CCATATAGGACAACATACTGGTAGGAATATTATCAAGTGAACGATA
ACGGTTCTTCTCCCAACAAGAAGTAAGCCTGGTGAGTGTATTTTGA
AGCTTTTTGGAGGGGCGGCCACGGAATAAGATATGAACACATACTG
GCAATTGGGCTGAGATCTGATTACATTGTCGACAGATTCCCATGAA
GGTGTTTGAGAGAATTTTGATTGAGTCGTTATACAAATTGCTGTGG
ACACGTACGCATAAAGTAGTTCGGGCAGCTATGTTTGCCACGTAGT
AGCTACAATATTAAGCATAGCAGACTAAATGCAATTGGCTGTGTTG
AAAGAAACTTCTGAACTGAGATTCAAGTAACGTGATTGTGTGCGCC
ATTTTACACAGGCTGCGCGGCAAATCAAAACCAAAGATACTTCTCC
GGAGGTATACGTGTTGCATTTTCAACAGACTATACGAGTTTTTTCT
GATCGTTGTATATTTAAGTGCAGCTTGGACTTACAAGCTCTATACT
AGGATAATGATCTCATTGGATTCAAGAGAAAGAAACTCTATACTGG
CGCCAAATTAGCAGTGTCAAATTTCGAAAAGGTGATG >YJL153C Chr 10 reverse
complement (SEQ ID NO: 4)
TTCTTTTTTTTTTTTTTTTTTTCTTTTTGGGAACGGGGGAAAAGGT
CTCACTAGTATATAAAGGAAAGAGAGTACAGCACGAATGAGCCAGC
TGCAGGAAAGGGGCTAGGTTAAAAAATAAAAACCCAGCAGAAAGCA
AACAACCAAATATAATTTAGAAATGGACAGAGACCATATTAATGAC
CATGACCATCGAATGAGCTATTCCATCAACAAGGACGACTTGTTGT
TAATGGTTTTGGCGGTTTTCATTCCCCCAGTGGCCGTCTGGAAGCG
TAAGGGTATGTTCAACAGGGATACACTATTGAACTTACTTCTCTTC
CTACTGTTATTCTTCCCAGCAATCATTCACGCTTGCTACGTTGTAT
ATGAAACGAGTAGTGAACGTTCGTACGATCTTTCACGCAGACATGC
GACTGCGCCCGCCGTAGACCGTGACCTGGAAGCTCACCCTGCAGAG
GAATCTCAAGCACAGCCTCCAGCATATGATGAAGACGATGAGGCCG
GTGCCGATGTGCCCTTGATGGACAACAAACAACAGCTCTCTTCCGG
CCGTACTTAGTGATCGGAACGAGCTCTTTATCACCGTAGTTCTAAA
TAACACATAGAGTAAATTATTGCCTTTTTCTTCGTTCCTTTTGTTC
TTCACGTCCTTTTTATGAAATACGTGCCGGTGTTCCGGGGTTGGAT
GCGGAATCGAAAGTGTTGAATGTGAAATATGCGGAGGCCAAGTATG
CGCTTCGGCGGCTAAATGCGGCATGTGAAAAGTATTGTCTATTTTA
TCTTCATCCTTCTTTCCCAGAATATTGAACTTATTTAATTCACATG
GAGCAGAGAAAGCGCACCTCTGCGTTGGCGGCAATGTTAATTTGAG
ACGTATATAAATTGGAGCTTTCGTCACCTTTTTTTGGCTTGTTCTG
TTGTCGGGTTCCTAATGTTAGTTTTATCCTTGATTTATTCTGTTTC
ATTCCCTTTTTTTTCCAGTGAAAAAGAAGTAACAATG >YPL033C Chr 16 reverse
complement (SEQ ID NO: 5)
GAAGTTACAAATAATAACAACAAAGTTAATAACAGTAGTAGCAACC
CTGATATTTCCACCAACTCGGTAGTTCATAACGCAATGCAATTTAC
AAATACAAACAACAATACAAGTAGCACTGTGGATATAAACGATCCC
AAAAATATTGCACCCCCTCCTACAACCTCTGTTTCAGCCCCAAGCA
CTCCAACCTTGTCCTCATCAAGTCAAATGGCGAATATGGCTTCGCC
TAGTACCGATAATGGTGACAATGAGGAGAAAAATGGTGGTAAGAAG
AAGAGATTTGGCCTATTCAAAAAAAAAAATAAATCAAAGAAATAAT
ATCCTTCGACAGCTTAATGAACACGCATGTAAATGTTTTGTAAACA
GTTTGTTTTTTCGTTTTTGTATCTGTTAATTTTCTATTATTTATTT
TTTTAGTTTTTTTTTGCGGTAGAATTGGGTTATCACTGAATCTATT
TTATTTTTTGGGTTGGTTTCTCCCTGTTTTCAGTTTGTTTGAATTG
GTGTTTAACAAAATCTGGAATTATTCACTATAATTATTTCCTCTTT
TTAGTTTGACCAAATAAACTCAACTATCTTGTGTTATTTCTTTGTA
TTATAGCTTTACACATATTTAACTATCCATACCTTTTACTTTTTGT
ATATACTGTTAATTTCATAAAACACGTTTTAGTACCAGAAAATCGT
GTCATAGCACCGTTTTAGCAGTGATTTTTAAAATTGTCCAAGGTGC
CAATAAATACTGGTGGGAACAACATTTTAATTAGAATAGCTAAAAC
CTTCTAAGAAAACTATCCCAGCGTAGAAAGAAAGAAATCTCTATCA
GCCGCGCAAGTGCAGTATCTCTTATTTTTGCACGGCCCCGTCGCTA
TATTGTCACACGCATTTGAATTAGTGACTCAATAATAATACAGATT
TGTGACGTATAAAGAAACCACAGAAAAACTGCGTATGTCCGAAGCA
CAGGTCCACTAGTAATAGAGCCCAAAAATCTGACATG >YOL058W Chr 15 (SEQ ID
NO: 6) AGGTTGCCACATACATGGCCAAGACCGGTAAGTCAGCCTTGGAAGC
AGAAAAGGAATTGCTTAACGGTCAATCCGCCCAAGGGATAATCACA
TGCAGAGAAGTTCACGAGTGGCTACAAACATGTGAGTTGACCCAAG
AATTCCCATTATTCGAGGCAGTCTACCAGATAGTCTACAACAACGT
CCGCATGGAAGACCTACCGGAGATGATTGAAGAGCTAGACATCGAT
GACGAATAGACACTCTCCCCCCCCCTCCCCCTCTGATCTTTCCTGT
TGCCTCTTTTTCCCCCAACCAATTTATCATTATACACAAGTTCTAC
AACTACTACTAGTAACATTACTACAGTTATTATAATTTTCTATTCT
CTTTTTCTTTAAGAATCTATCATTAACGTTAATTTCTATATATACA
TAACTACCATTATACACGCTATTATCGTTTACATATCACATCACCG
TTAATGAAAGATACGACACCCTGTACACTAACACAATTAAATAATC
GCCATAACCTTTTCTGTTATCTATAGCCCTTAAAGCTGTTTCTTCG
AGCTTTTTCACTGCAGTAATTCTCCACATGGGCCCAGCCACTGAGA
TAAGAGCGCTATGTTAGTCACTACTGACGGCTCTCCAGTCATTTAT
GTGATTTTTTAGTGACTCATGTCGCATTTGGCCCGTTTTTTTCCGC
TGTCGCAACCTATTTCCATTAACGGTGCCGTATGGAAGAGTCATTT
AAAGGCAGGAGAGAGAGATTACTCATCTTCATTGGATCAGATTGAT
GACTGCGTACGGCAGATAGTGTAATCTGAGCAGTTGCGAGACCCAG
ACTGGCACTGTCTCAATAGTATATTAATGGGCATACATTCGTACTC
CCTTGTTCTTGCCCACAGTTCTCTCTCTCTTTACTTCTTGTATCTT
GTCTCCCCATTGTGCAGCGATAAGGAACATTGTTCTAATATACACG
GATACAAAAGAAATACACATAATTGCATAAAATAATG >YOR255W Chr 15 (SEQ ID
NO: 7) CTTAGTATACGATTGTGTTCTTGCCCACCATCTACTAACAAAATAT
GGCAAGATTAAACCTAGTAGCGCAACATAACAAACCACTAATAATG
GAGATGCACTTCCATCTACCAAAAATCTTGGTAGAGCGATACCATG
TGAAGTAGATTGTGGGCCATCTGGATGACCGTATTTCAAATAGTTT
TGCCTAACCAATTCGTCAGTAAGGGATTCGTAAGCCTTCGTAATCT
GAACATAAGTTTCTTCCATCACACTTTTCTCATCAGGTGTTAGGCC
CTTTGCTAATTTATCTGGATGAAATTTAACAGATAATTTTCTATAA
GCAGATTTGATGTCTCTATCGGAAGCACTAGTAGAGATACCAAGGA
TTTCATAAGGATCAAATAATTTTGTAGCAGCGTCTTTAATCGCGTC
ATTACTATTAATCCTTTGCAGAAGAATTGCAACTAAGATCCAACCC
ACAATAATTATAATATTTCTCCTGCTCCATATTTTGGACTTCTTAT
TACTATTTTTATCAAACTTCCTTCTAAATTGTTTGATTTCATCACT
GGTGTATTCTTCATTCAAGTTCTTGAAAACTTCCTCATTAAACTCC
TTACTCTTCCCTGAATTCCCATCTTCAGCATTGGCCCCAAAAAAAA
TTTGGTATATTTGAAGCAGTGTCATAGGCCCGACGACCATCAAGAG
CCCCGTTAAAATGAAGGACGGCCACGTCTCACTAGCCTCATCATAC
TCGTAATTTGTAGGCATTGTGCTGTAATATGCAGTATATTTATTTT
GTTGCGAAGGCCTTCTTTGGTCAATTATAACTGACTATTGTTCCAG
TCGCATCTTTTATTCGTTCAGTGATCTTCGACGTGTTCAAGACAAA
TTTTTCAGAAAACGCCTTTCCGTAACGAGTGACCAATTCGCGTTAT
AAGAGCGATTGTGTCAGTAATGAAGTCAGATTGTGACATTATAAAG
CTAAGACGTTAATAACTCAAAATAGAAAAGAAGCATG >YDR250C Chr 4 reverse
complement (SEQ ID NO: 8)
AAATAAGAGGTTTGGCCAAAACGTGGTCATCCAAGGTTGATGGATT
CTTCTCTTCTAATATTATTAATAATGGATCGAAACATATTTGAGGC
AAAGCTAGTTCCCATTCTTTTGCCAAGAAAGCGGAGGGAGAATGAT
CGTAAAGAGTCACAACGTCCCTAGGAAACAGTTTTTGGAACAATTT
CTGAAATGTATTCAAAATAGGAATAATATCACTTGAATAGCTGTTT
TCGTGAACGGATGTCGTTTGACCGATCGAAACAGAAAAGGATTTGG
CATTTGCAGTCGTGAACTGTTTGTATGAGCCATTGTCCAACCGTCT
TATGTCATAATCGGTGAAAATGCTGAAAATATTTGACAGCGAAAGA
TCAACTGAAGCCTTGATGAATGGCTCCAAATATATAAACCCGCTAC
TTTCAAATAGTATCTTTGTTGTCTTATTCATCATAGGTTTTATATC
TCTCAATACCTGCGGGATTTCTTGCAGTGAAGACGCACTCATTATT
ATCAAGTCGAATACCAAGCCACCATTTTCATTATTAAGGTCTACTA
AATCTAGGAGCGATTGGAAATGGTTTTGAATCTGAAACCGTTCCGT
CCCATAAAATAGTGAACTAACCTCGTAGTTGGCATTTTTTGAGTTG
TTCACAAGATAAAATTCTATATTTTTGGCATGTTGAAACCTGGATG
TGTACAGAATGAGATTTGGGTGATCTCCGCATACCAAAACCCTTAA
TGCAGATGTCATTATTCAATATACCTATCGCGAGATATGCCGGGAA
TAAAATGTAGGAAACAAAAAGAAAAATATATGCACAGTAGTTCAAA
CAGAATAGAAATGACGGAGAGCAAATATTTGGGTAGAATCACAGAC
TTTAGTGTTGGTTACAAGCTTTTCGAATGGATGTAGTTTGTAATCA
AAGGACACTAATAATAGCGAAAAAAAAAAGGGCAAGTATGTTGGCA
GTCGCCTCTTTTTCAATTTAGCGGTAAGTTTTTAATG >YDR446W Chr 4 (SEQ ID NO:
9) CGCAAATGATGAGAAATAGTCATCTAAATTAGTGGAAGCTGAAACG
CAAGGATTGATAATGTAATAGGATCAATGAATATTAACATATAAAA
CGATGATAATAATATTTATAGAATTGTGTAGAATTGCAGATTCCCT
TTTATGGATTCCTAAATCCTCGAGGAGAACTTCTAGTATATCTACA
TACCTAATATTATAGCCTTAATCACAATGGAATCCCAACAATTACA
TCAAAATCCACATTCTCTACATTACTAGTATATTATCATATGCGGT
TTAAGAAAATGGTATAAAGATTAAGAAACAGTCATGAAAATTTAGT
GAAGCTGAAATGCCAGTATTGATAATACGATAGAATAATGAATGAC
AAAGTATATAAGGAAGATGAAGTAACATTATTATGGAGAACTATCG
ACTCCCTTTTGTGAATTTCTATATCCTCAAGGAGAATTGCTTGTAT
ACTATGTATATATAATATTATAACCCTTGACAACAATGGAATAATT
AGTCCTTAAACGTTAGACTGGCTGGGACAGCACGACCTCTGTTGGC
TAATTTTACTTTTCCCACAACCGTAGATCTTGATTTCATGACTGTT
TTCGTGGTAATGTCCGAATGATACATATATAAGTTATATGCTTTCA
CTTTATAAGTATTAGCGTTTTTACTCACCATCCACTGCTTTGAGAA
GATGTGGTCAGCAACCACGCAATACATCACATATTTTGGCACGGGT
GATGTACCGATCTAATGGCTAAGCTTCCTTTTGGAAGCTGATAGAG
ATCCAGATTTGCATCTCTATCGCTATTGTCGTCATCGACACTGCTC
ACTTAGTGCTGCAGGTAAATTCCGTTTTTTTCCAAATTAATATTTA
TGAACGTTATGATGTCAAGTTTTTTCAAGAAGTAATTATCCGCGAA
AAAAAGAATATAAAAAATACAAATGTGCATAGATCCTCACATAGTA
TACAACTAAAAAGCAAACAAAAGAACATCCTCAAATG >YDR536W Chr 4 (SEQ ID NO:
10) CAATGATTCTGAAATACTCCTTTTACAACCTTTGCAAAGATAATGT
CTTTCAGTCTGATATTACGAGCGACCTAGAACCACTAATCCATATT
CTTCATTCAACTTACTCCATTTTCCTTGGCAAACAATGCCCCACAA
TCATATACGTCATAACTATAAGGGATATGTCTGGAATGCGGCCAAG
ATAGAATTAAAGGGCTGCAGAACACCACTACTGATACTCATTGCCA
AGGCTAGGAGGCACCATCCGTTTCATTTTCTTTGAGGTAAGCCAAT
CATGAAATAGTATACACATCCATAACGGACGTACGGACGAAATAAG
TGCCGTTGTCCCACTATTCCACCGCATTTGGCCCATTTGGCTCACT
TTGACTCAACTTGCGTCATTTTAACTGATATGAAGGGTCCGACTTT
GTCCTTTTTCGGCCACCGCATACCCCACGGCGATGCCTCCGCTACC
TGCATTTGAGTAGCATCTCCGTTTCGCGGGGTATTCGGCGCTACGT
CGCCTGTTCGAGCGGCTCTGTTCGTTGCATGAAACTAAAATAAGCG
GAAAGTGTCCAGCCATCCACTACGTCAGAAAGAAATAATGGTTGTA
CACTGTTTCTCGGCTATATACCGTTTTTGGTTGGTTAATCCTCGCC
AGGTGCAGCTATTGCGCTTGGCTGCTTCGCGATAGTAGTAATCTGA
GAAAGTGCAGATCCCGGTAAGGGAAACACTTTTGGTTCACCTTTGA
TAGGGCTTTCATTGGGGCATTCGTAACAAAAAGGAAGTAGATAGAG
AAATTGAGAAAGCTTAAGTGAGATGTTTTAGCTTCAATTTTGTCCC
CTTCAACGCTGCTTGGCCTTAGAGGGTCAGAATTGCAGTTCAGGAG
TAGTCACACTCATAGTATATAAACAAGCCCTTTATTGATTTTGAAT
AATTATTTTGTATACGTGTTCTAGCATACAAGTTAGAATAAATAAA
AAATAGAAAAATAGAACATAGAAAGTTTTAGACCATG
>YOR255W Chr 15 (SEQ ID NO: 11)
CTTAGTATACGATTGTGTTCTTGCCCACCATCTACTAACAAAATAT
GGCAAGATTAAACCTAGTAGCGCAACATAACAAACCACTAATAATG
GAGATGCACTTCCATCTACCAAAAATCTTGGTAGAGCGATACCATG
TGAAGTAGATTGTGGGCCATCTGGATGACCGTATTTCAAATAGTTT
TGCCTAACCAATTCGTCAGTAAGGGATTCGTAAGCCTTCGTAATCT
GAACATAAGTTTCTTCCATCACACTTTTCTCATCAGGTGTTAGGCC
CTTTGCTAATTTATCTGGATGAAATTTAACAGATAATTTTCTATAA
GCAGATTTGATGTCTCTATCGGAAGCACTAGTAGAGATACCAAGGA
TTTCATAAGGATCAAATAATTTTGTAGCAGCGTCTTTAATCGCGTC
ATTACTATTAATCCTTTGCAGAAGAATTGCAACTAAGATCCAACCC
ACAATAATTATAATATTTCTCCTGCTCCATATTTTGGACTTCTTAT
TACTATTTTTATCAAACTTCCTTCTAAATTGTTTGATTTCATCACT
GGTGTATTCTTCATTCAAGTTCTTGAAAACTTCCTCATTAAACTCC
TTACTCTTCCCTGAATTCCCATCTTCAGCATTGGCCCCAAAAAAAA
TTTGGTATATTTGAAGCAGTGTCATAGGCCCGACGACCATCAAGAG
CCCCGTTAAAATGAAGGACGGCCACGTCTCACTAGCCTCATCATAC
TCGTAATTTGTAGGCATTGTGCTGTAATATGCAGTATATTTATTTT
GTTGCGAAGGCCTTCTTTGGTCAATTATAACTGACTATTGTTCCAG
TCGCATCTTTTATTCGTTCAGTGATCTTCGACGTGTTCAAGACAAA
TTTTTCAGAAAACGCCTTTCCGTAACGAGTGACCAATTCGCGTTAT
AAGAGCGATTGTGTCAGTAATGAAGTCAGATTGTGACATTATAAAG
CTAAGACGTTAATAACTCAAAATAGAAAAGAAGCATG >YDL243C Chr 4 reverse
complement (SEQ ID NO: 12)
CATAAGAAACAAATTTTGTAAGTTTCTTGTGCCTTTTGAGTTTGAA
ATCATTGACCAGAACAATGAGAACTCTAAACACGACAAATGGATCT
TAAATCAATTTTCTTGAAAACATCTTCCAGTTTCGATATTCGCACA
TTTGCCGGTAAAATAGTGCAATCTGCCTATACCTTTAAAATGGGAT
CTTGAACGCGTACCTGAAGGTTTTTTTAAATAAGGCCGAACCTAAA
TGTTGCCTTCAAAATTAACCTGTATCAAATTTTGAAGCCGAAGTTG
GTATCCTCACATTGTCTAACCTCATGGTCTCCTAAAGTCCGGCGAT
CACTTTCCGTTGCGTGACTATTGATGATCAGACTGTCAACAATGTC
GTTCCGGGCCTCCATAGCTTGAAGAATTTTCGTTGACGGAACTAGA
GGAATGGCAGATGGAGATGACCTTAGTTTCATGTCGCCTTGATAGT
TATCAAAGAGTACAGAAACAAGACGTCTCTTGCTCAATAATAAGGA
TTGGATCAGGTGCTTACAAATTTTATTCGTCATACAAACAATTACT
AATATTTTTTTAATATTAAGCGTACCCCTATTTGATTGGCTCTCTT
CTAAATTCATACGTTTTAAAATCCGCTTTCAACCGTCAACATAAAA
TTAGTAAGCAGACCTCGTTCCAAGACCCAACTAAGTTGCACTAAAT
ATCCTCGAACAAATTGGGGGCAGATTTTTGAACCATATAAAAGTCA
TGACAATATTTCCAGATCAAGGACAGTTGCAGTTCTTCCTTTCATC
GGCCAGCTGACGAATTGACTGAGCGGCTTTGTATTTCACTAGAATA
CACTTCCTATAGCTACAAAATATTATTCAATAATATTAGTGAATTA
TTTAAACCTCTACCTGAACCACCTACCGAGTTGGGACGTCTCAGGG
TTCTTTCTAAAACTGCCGGCATAAGAGTTTCACCGCTAATTCTGGG
AGGAGCTTCAATCGGCGATGCATGGTCAGGCTTTATG >YDR256C Chr 4 reverse
complement (SEQ ID NO: 13)
AGAACATCCAAATTCGGAACTACTAAGAAGATATGGGTATGTTGAA
TGGGACGGTTCGAAGTATGATTTTGGAGAAGTGTTACTTGAAAATA
TTGTCGAGGCGTTAAAAGAGACTTTTGAGACGAATACTGAATTTTT
GGACAGGTGTATTGATATCTTACGCAATAACGCCAATATTCAAGAA
TTCTTAGAAGGTGAAGAAATAGTACTAGATTCATATGATTGTTATA
ATAATGGTGAATTGTTGCCTCAACTAATACTTTTGGTCCAAATCTT
GACAATTCTTTGCCAAATTCCAGGTTTATGCAAACTGGACATAAAA
GCAATGGAAAGGCAAGTGGAGAGAATTGTAAAGAAGTGTTTACAAT
TGATAGAAGGTGCCCGCGCCACTACAAACTGTAGTGCCACATGGAA
ACGTTGTATTATGAAGCGTCTAGCCGATTACCCCATAAAAAAGTGC
GTTTCTATCGAAAAACCTTCGAAAGGAAACTCATTAACAAGGGAAG
AACTAAGAGATGTTATGGCTCGGAGAGTTTTGAAAAGCGAAATAGA
TTCGCTGCAAGTTTGTGAAGAAACCATCGACAAGAATTACAAGGTT
ATTCCTGATGAAAAGCTGCTAACTAATATTTTAAAGAGAAAGTTGA
CAGAGGAAGAAAAAAGCTCTGTCAAACGTCCTTGCGTGAAGAAGTG
AGCGGTTGTTCTAACCACTATTTAAAGCCGCAATTAGTAATGCAAA
AAGTTGGCCGGAATTAGCCGCGCAAGTTGGTGGGGTCCCTTAATCC
GAAAAAGGACGGCTTTAACAAATATAAACTCCGAAAATCCCCACAG
TGACAGAATTGGAGAAACAACCAGTTTTGATATCGCCATACATATA
AAGAGATGTAGAAAGCATTCTTCACTGTAATGTCCAAATCGTACAT
TTGAATTTCTTGTAGGTTTATTTAAAAGGTAAGTTAAATAAATATA
ATAGTACTTACAAATAAATTTGGAACCCTAGAAGATG >YFL020C Chr 6 reverse
complement (SEQ ID NO: 14)
GTTGAACTTTATTCAAATATCATCGAAGTAAGAAGATATACTACTA
AAGACTCCCTTTGCTCTATATTCGAGTCAGGCTCCACCAGTCATTT
CGAAATTAACCAGTTACAGGTCAAAAGACTAAACTTACTGCAAAAC
CAATTTGCTTCTGTTTTCACTTCTTTCCACCCCAAAGATAATACAA
AGGGCATACATATCAATTTTTTTTCACCTGTCACTAGGATAACTGA
CCTTCAATATTCCTTTTTTTATACGAATCAGATACTGTTCGGTACA
CGATATCTAATTAAAATGATTCAAAACTTTGTAACAGGTAAAGTTT
TCACTAGAACCAATCAATCCAAGTGAATTAGGGGAAACCATAGTTG
TTGATTTGTAGAAACCTCACATTGTACATTGTTGGTTTGTTGGGCA
TATCAGAACGAGAGATTTTCCAACATTCAATATACACTAAACCCTA
TGACGAGTCCCACAGATGGCGTAAGGTTTTTATGATTTCAGCAGGG
TACGACGACTAGTACCATATTAACATTTTTTAGTGTTTCTAATTTG
GGAAAAGGTCCGTGTTTTTTCTCCTAGCAACCGTTTAGTGCCAAGG
GTTAGGCAATTGAACGAGGCCAAGACAATATTGGCTTTGCTTCTAT
TACTTGGCTAACATTGTGTCTGCAGGTCGAAAGGCACCTTTACTGT
AAGGAACATTCTTGCGCTCTAAACATACGAAGATATGGGGAATATG
AAGCGTGTTTCTTATACGAAGTGCAGCATCGTTCAAGGAAAATACA
CCCCCATAGTAATAATGGCTAAGTGGCCAGGAATTAGAATATGTGA
GATATGAGTGCAAAATGAGTGACCAGTAATAGCCTGTTTGGGATGT
AATTGCTCAAAAAATTTATATAAATACAGCGGTTTGATCAGCTTTG
TTTGAGACATTTCTCTGTTCTTTTCCTTCCAGTTAAGCTTATATCT
CCACTAAGCAACAACCCAAAAAACAACAAATACAATG >YPL205C Chr 16 reverse
complement (SEQ ID NO: 15)
CAGACTTGTTGTCTGTGCTGTTACTTGCACTTGCTGCATTGCTATT
TGCGTTCAAATCACCTTTTTCGATGAGGAGGTCCCTTTGCTTCTCC
ACCATCGCCTTTGTGTAACGCAACATTGTCCAATCGAACAAGTGGT
CGTTGTGATACTCTAGTTTAATACTCAGATCTTTAAACAGCCTTGC
CAAGAACAAATAATCTGGCTTCTCATCGAATTTCAAATTCTTACAG
TAAGCCATATATTCTTGAAACTCTAATGGTAAACCTGAACATAGAG
TTTCCACGCTAACGTTTAATTTCTTTTCCATGATACGATCATACTT
TTGTTTCTTGGTGGTTGCTTTCAAACCCTGCCATGGCAAAGAACCC
TTACAAAAATAGATCAAGACATAACCTAGTGATTCTAAGTCATCTC
TTCTACTTTGCTCTATTCCAAGATGCGTATTGACACTTGCATAACG
AGCTGTACCTGTCAAGGACTTGTTCTCCCTGTAAGGAATATGACGA
TGTGTGTTGAAATCTCGGTATTTCTTTGATAGACCGAAATCAATAA
CATGAACGGTGCTACCACGGCGTCCTACCCCCATTAAAAAGTTGTC
TGGTTTGATATCTCTATGAATGAACGACCTTCCATGTATATACTGA
ATACGGCAAAACATTTGCAAAGCCAGCATGATAACCGTCTTAAAGG
AGAACCTTCTGTGACAGTAGTTGAATAAATCTTCCAAAGATGGGCC
TAGAAGATCGATGACCATAGCATTATATTCACCCTCTCTGCCAAAC
CATCTGATGAACGGGATTCCCACACCACCGCTTAAGTATCTGTAGA
CGCGGGACTCATAGTCCAATTGAGGATGTCTGGACCTGATCGATTC
CAGCTTGATGGCTACTTCTTCACCACTAATTAAGTTCGTGCCGTGG
TAAATGTCACCAAAGGAACCACTCCCAATCTTCCTGCCAATACGAA
ATTTCCTTCCTACTCTTAAGTCCATCTCTTTTTAATG >YGL205W Chr 7 (SEQ ID NO:
16) TCACTCAACCACCTCCAAAAAATAACAGGTTCATCTAAAGTAAAAG
ACTTTAACTTGCTCTTAGTTTCCAAATTAAATATCTGCACGATAGT
ACCATTTGCTCTAACGGAAATAACCATCTGAGATGGATGCATGATA
GCAGAATCACCGCCCATATTCTTCCTTGTCACTTCATTGCCTTTGG
CCAAATCCACGATTGCAACAGAGTTTGTACCGTCCTTTGTTTCTCT
AACAGTGACGAAGTGGTCACTCTCGAAAGTAGTTGATCTGAAGTCA
AGGAATTGAGGGGAAATTCCTAAGGACATCAGATCGACCAATTCGG
TAAATTCAATGGGTAGGTCACTCATTGGTTAGAACTTTCGTGATAA
TTTATTTTTATAGTTGAATATCTTCTTTCTCTCTCAACTCTGATCC
GGATTGTCGAGGTTTCAATAAGTTACTCTGAACAACTAATCAAAAT
ATCTCCTTATTTCTGTAGATTCCTTCAGTTCCACTTTTTACTTTTC
TTAATTCTCTTTGTATTTATTCCTAGCGACGAAAAATGCGAGATCT
CGACCAAAAAAAGGGGGTAGGGTAATAAAATTAACCCTATTATTTT
TTAACTTTAAAACCTATAATGTGCTAATATTTTATTATAAACCTCC
TTTTTTTGCGTTCAAACCCTGACACATTTTAAGCCCTATATTTACG
GTATTAGTTGATTAAACTCCGAAGCGAAAGGAATTCGGTCATTAGC
GGCTAATAGCCGTTGGGGTAAATCACCTACAAGCAAGTACACAAGA
GAACGTTGGCGTTGTTAAGTCAAAGCACTAATACATTGGGGCTTTA
AGAGTGTTTATAAAGGTCTAACCTGTAAAAATTATTTAAACAACTT
GAACAGGCCTTAAAGTTTTCCTCATTCCGCTCATCATCACTAATAT
TGCTCTCCGTTTTTGAATACACACTTGACACTAATAAGTATCACAG
AAAAAAAGAAAATATAATAAATTAGTATTGCGATATG >YGL117W Chr 7 (SEQ ID NO:
17) AAAACGGCTCCTAGTTATCGTCAGTTTTGATAATTAGTAACTAGAA
TATAGTATCACCAAGAACAGGCCAGGTAATATACTAGAAGTTGTGT
ATAACTACTAATATTTTTATACATATAGATGCCAAGTCAAGGTCCA
ATTAGTAGATTTTTCACTCTACCTTGCATTATTACCTGAAAGATCG
TTTGTGTTTCCTTAAATTCGCCAGAGGTGCAAACGAATGTAAATAT
CTACATCAATTTACGGTTTCACGCCCGTTTCGTTTCTAATGGCAAC
TCATGTAAACTGTGATAAACCCGTTATTTCAGTTTTATTTTCTGCT
AGTATAAAGGATGATAGTAGCCATTTCGCTGGAATATGTATCGGGT
TTAAAAGATGGCATAGTTCGAAACATTTTCGCACAAGATTCTAACT
GTCTTCGAACGTCGTAGAAACCGCAACGCAAGAAATGACAGCGCAA
TAAAATTTATTTTACTAACTGATAGAACAAATGTAAAAAATTTCAT
AATATCAAGTAGAATTCTTTATTCTCTTTGTGAATTCCTAAACCCA
TGAAAGGGACTTGTAGTATAAGCTGGATACCTATTTTCATTGGCTT
CGTTTCCAAATGACACAAAACCATCACAAAAGGAAATAGTCCTTCC
ATAGTTACCCAATCATTCTGTTGCATATTAAATGAACAGCTCCTTG
TTGATAATGCCAGTGTGGCATATTCACCGCTGCAGTTATTTCTTTT
CATATATAATGAGTCATTTGTTTCTGTCATCACTTTTCTATACTTT
CTCTTCCCCGCGTGTTTTCCGTACACCAACAATATATGCCATAATA
CACGTAACATTTTTTTATAAAAAGAAAAGGTAAGTGATATATATAA
AATAGCGCCATGAGTAGGAAACTTTTCTGTTACTGCAGATATGTGC
CAGACTGGCTCAGGTGGCATAAACACAGATTAATAGTATTGGCGTT
GCTGAAATAAGAAGATCTGTAACAATATCTTACAATG >YHR029C Chr 8 reverse
complement (SEQ ID NO: 18)
TCTAAAAATGTTCAGGACTACATACATCAATTAGGTTTCATTCCAA
AAGTACCTTTTGTCAATTTATACCCAAATGCCAATTCACAAGCATT
AGACTTATTGGAGCAAATGCTCGCGTTTGACCCTCAAAAGAGAATT
ACCGTGGATGAGGCCCTGGAGCATCCTTACTTGTCTATATGGCATG
ATCCAGCTGACGAACCTGTGTGTAGTGAAAAATTCGAATTTAGTTT
TGAATCGGTTAATGATATGGAGGACTTAAAACAAATGGTTATACAA
GAAGTGCAAGATTTCAGGCTGTTTGTGAGACAACCGCTATTAGAAG
AGCAAAGGCAATTACAATTACAGCAGCAGCAACAGCAGCAGCAACA
GCAACAGCAACAGCAACAGCAGCCTTCAGATGTGGATAATGGCAAC
GCCGCAGCGAGTGAAGAAAATTATCCAAAACAGATGGCCACGTCTA
ATTCTGTTGCGCCACAACAAGAATCATTTGGTATTCACTCCCAAAA
TTTGCCAAGGCATGATGCAGATTTCCCACCTCGACCTCAAGAGAGT
ATGATGGAGATGAGACCTGCCACTGGAAATACCGCAGATATTCCGC
CTCAGAATGATAACGGCACGCTTCTAGACCTTGAAAAAGAGCTGGA
GTTTGGATTAGATAGAAAATATTTTTAGGACAAAAAACTATAAGTA
ACCGGGGAAGTATAGAATCACCATAGATGTAAGCTTACAGACAATG
TGTATATATGATGTATATGAACGTATACAAATATATATATATATAC
GTGCTCTTGTTGTAGCTCGTATATCAAATTCCTCCTCCGACGCTTA
TCTTAATCGTACTCCGCGGAAGTTTGTTATCGCCTCTTGAATTCTT
TCTTTTCGTTCATTTATGATTAGTCATCTATAGACAATATTCATTA
TTTAAGCACCTAGAATACTAAACTAAATGTCTAAATATGACACAAG
GAAGATAAGATAAAAAAAACCAAGCGCTTAGAATATG >YML116W Chr 13 (SEQ ID
NO: 19) CATCGGGATCTTCCGACTCGTCATCAGACGTCGAGGTCATTATGCA
CTCCCCGAGCGATCCTGAATACGCTTTAAAATCACAAACTCTAAGA
AGCTCGAGTCAGACCGTCATTAATAGTAAGAGACCAGTAAAGATAG
AAGACGAGGAAGAAGCCGTAGGAATGTCACAGCTCAATTATAGGTC
GTCATTAAGACAAGCTCCTCCAAGAGCTCCCTCAACTTTGTCATAT
AATCACTCGAAGAACAACGAAACGCCAATGCAAGATATTTTCACAA
ATGGCGAAACAGCAAATAACAGAAAGAAGAAGAGAGGATCTTTTGC
AAGGCATAGAACGATACCAGGATCTGACGTCATGGCCCAATACCTT
GCACAAGTGCAACATTCGACATTTATGTATGCAGCCAATATTTTGG
GCGCCTCTGCGGAAGACAACACGCATCCTGACGAGTAGTATAGCTG
TGCTGAGCCTGAAGTATAATGCATATCCATCGGACTATTTAGACAA
AGCCCAAGGAAGCCTAAGGCGGCCTGCCAAGGTGTTTCTCCCTTTT
TTTCTCCGATTTCTTTGTATAAACTAAATAATATAGTGATTACTAA
TGGAATGGCGGTATTATGCACCTAACCTGTTCATTCTGCCACAGAT
TACGTAAGCGATTTATTGCCGGCACTTGTTGTCTTAATGACCGAGT
CACCAATGTGGAACGATAATTTTCTCTGACTCAAACCTGTTAATTT
TTTTCTACTTCGTTTCGTTAGCGACGACGTCAAGCCGCAGGATCCT
GTCTGCCTTGACCTCTTCTCTACCTTCAGAACTTACACTTTTCAGG
TAAGATGACCTTTATATAAAAGTTGACACTATTACAGTTGTTTATA
AACGTCTGAAGAATGAGACGTTTTTATAAAATGAATAAAATGCATA
TTCTAAGTTTAAAACAACATTTTCAAAGTGTACGATTGTAAAAAGA
GAGGCAATTAGAGAATCTCAAACAGGTAATAATAATG >YJR109C Chr 10 reverse
complement (SEQ ID NO: 20)
GAAATCCAATGAGAATACCGCATAATCTTTCCCAATTACCTTGATA
TTTACAAAAGACCATAGATCCTTACCCTGGTAAAGCTTGTAATTCT
GTGAATCCGTTCCCTCCAATTGGATTTGATTAGTGGAAGTCAACTT
GGACAGTAATGTACTACCCGGGTTTTTATGCACTCCATAGATCAAA
GGATATGGAAACCAAAATTCCGTCGACAATTGTGGTGACTCAAAGA
TCAAATGGTGAGTAGTCAAATGTAAAGTACCTTGTGTGGCCGTACC
TCTTCTATGCAGCACTACATTAGATACTTTGGCAATCTTGATGTAC
TCCATCTTTGCGGAATGGTATACTATTTCTTTTCCCTCTTTTTAGC
ACTATTACACCCCGCCCACAAAATAAAAATAATAACGACGACCTAA
TCTCACCAAGTGACCCTTGTAAAACCTCCTTTTCTTTATAATGTTT
CTTTTCTTACTAATATTTGGTACATTTAGGGTAGTGATAAAAGAAT
GGCAACATTGTTATTATTGTGAAAAATGAGGAAAATAGAAAATCAG
AAACCCTAAAAAGTGATTTTACCCTATCAGAAATATTCAAATGTCC
TAATTAAAAATAGTAAATCCCCTAAACATTCAGATTGTAAACTAGG
GTTGAGAAAATGACTCATCCACCACTGTCTTCTTTCCTGCGGCATT
CTATAGATTATTGTGAATGACTCTTATTGATGAGATGGCAATAACT
TTTGAATATCAGAGATAGGAACCTCCATGTCGTAACGATTGTGTCA
CCTTGAGTAAGCATCGAGAAAATCCAATCTTTTTTTTTCCGTCATA
AGCATTTCTGCCATGCTATTTGTATATATATAATTACTAATACGTC
TTCTATAGTATGCCTTATCTCTTTTTTGAAGCGCTATTTAAGTTTA
AGCATCGAAAAACTAACATCTATAGTTAAAATTAGTTCTATAAAGG
AAGAGCAATACAGTACATAGACAGGAAGAAAAGAATG >YCL030C Chr 3 reverse
complement
(SEQ ID NO: 21) GAATTACGAGAAGCCCAATTGACCATCGAAAAGCTACAAAGGAAAC
AACTACACTACAAAAGGCTACTCGATGACCAAAGAATGGTCCTCGA
AGAAGTGCAACCGACTTTTGATAGGTATGAAGCCACAATACAAGAA
AGAGAGAAAGAGATAGACCATCTCAAGCAACAATTGGAGCTCGAAC
GCAGACAGCAAGCCAAACAAAAGCAGTTTTTTGACGCTGAGAATGA
ACAGCTACTTGCTGTCGTAAGCCAACTACACGAAGAGATCAAAGAA
AACGAAGAGAGAAATCTTTCTCATAATCAACCCACTGGTGCCAACG
AAGATGTCGAACTCCTGAAAAAACAGCTGGAACAATTACGCAACAT
AGAAGACCAATTTGAGTTACACAAGACAAAGTGGGCTAAAGAACGC
GAACAATTGAAAATGCATAACGATTCGCTCAGTAAAGAATACCAAA
ATTTGAGCAAGGAACTATTTTTGACAAAACCACAAGATTCCTCATC
GGAAGAGGTGGCATCCTTAACGAAAAAACTTGAAGAGGCTAATGAA
AAAATCAAACAGTTGGAACAGGCTCAAGCACAAACAGCCGTGGAAT
CGTTGCCAATTTTCGACCCCCCTGCACCAGTCGATACCACGGCAGG
AAGACAACAGTGGTGTGAGCATTGCGATACGATGGGTCATAATACA
GCAGAATGCCCCCATCACAATCCTGACAACCAGCAGTTCTTCTAGG
CAGTCGAACTGACTCTAATAGTGACTCCGGTAAATTAGTTAATTAA
TTGCTAAACCCATGCACAGTGACTCACGTTTTTTTATCAGTCATTC
GATATAGAAGGTAAGAAAAGGATATGACTATGAACAGTAGTATACT
GTGTATATAATAGATATGGAACGTTATATTCACCTCCGATGTGTGT
TGTACATACATAAAAATATCATAGCACAACTGCGCTGTGTAATAGT
AATACAATAGTTTACAAAATTTTTTTTCTGAATAATG >YLR338W Chr 12 (SEQ ID
NO: 22) TGATGCAGTTGGAGTCGGTAGCGATGTTATAGGAGGAGGAGGTGCA
CTAGGAGGCAAAGGAGGCGCCATAGCATCCGTATGGGGGGGTGGTG
CGTTAAAAGCATTCGATGATGTTTGCTTATGTAATTTGGATTCAAT
TTCATCCAGAAACGACATACCATTTTGGCTACTCTGTTTAGGCGGG
TTTATAGATGGAGAAATAGGAGAAGGTCTTACGTTATCATCCTTAG
ATGAACCTATTACATCCTCTTGTTTATCTTGGGTAGTGTATCCAGT
ATCACCACCAACCACGAATCTATCATCTCTCTTCTTTTGGATCTCA
GCAAGAAATGGTAAGGGTCCCCCAGGTGTTACGGCCGAGGAAGAAC
TGGATGAAGATATTTTGGATTGCTTACTGGACGCTGAGGGTTTGTT
TGTCGTGAGTGTTGGAGGAGGAGGAGGAGCCGATGGTACTGACGTT
GCAACCGAAGGAGGAGGAGGAGGTGGTAGTGGTGCTAAAGGAATAG
ATGATGCTGACAAGGCAGAGGATGTTGAAAAAGCACCAGGTGGTGG
GGGAGGAGGAGGTGGTGGCGCGGGCATTGACGATGCTTTGTTAGGA
GGAACTGATGTAGTATTTGGTAACGGTGGTGCTAAAGTTGGGGGAA
CCGGGGTTGCTTTTACGCTGTTAGTCGACGCAGAGCTCATGGCTGC
TGGTAGTGGTGGTGGAGGCGGAGCTGGAGCTGAAGTTGCTTTTTTA
GGTGCAGAGGTCACATTTGGTAACGATGGTGGAGGTGGTGCAGTAG
GCGGTAATGGCGGAGCATGTGATGTAGGAATGGGCGGAGCAGATGA
TGGAAGTGGTGGTTGTGAAGGAGATTTATGATTTTCGGTTTGAATC
TTTGTGGAAGAAACGCCCTCGACGGCTCCTCGTTCAGACCTCCTGG
CATTAATTTCTGCCAAAAATGGCAACCCGCCTGCAGGAACTTCAGA
AGATGGTGGTGAGACTGCGTTATCTGTAGGTTTAATG >YIL066W-A Chr 9 (SEQ ID
NO: 23) TGAATAAGCCTGTATAACTGCGCATGTAAGAATAGTGGCTGCAATA
GATCCCACAGCAGTCAGTGCTCCAGCCTGCTCTTTCAAATCCGTCG
TCGTAAGTCTATCAACAGGTAATCGGTCAAATGTGACATTATTTGA
GATTCCCAACCAAAATGTGGGGTACCAAAGTATTACTTTTGCTATG
GGCGATCCATTTTCTAGTGTTCTTTTCACCGCACTCTTCCAAACGA
CGTATAAAACAAAGCATAAAGGTAAAAGTCTTTGAAATCCCACACT
GAAAAGCTCATACACGCTAGCTGGGTCATGTGCATCCGTTAGTAAT
GGAGGGTCTAGAGATAATACAAGAGTCCAATAGCCCACTATCGTGT
TTATCCAGTACCCGTAAATACTGTCATACAAATAAAATATTGGCAG
TCCAAAAAGCATATTCAATGCGACCACAGCTGCTCGAGGATCATAA
CAGCCCGACAGGATACCGCCTTGTATATCTCTGAATGAATATGAAC
CGGGAAAAAATGAATTGAATGCTACTGAAAACCCAGTGTTATACAT
TCCTTTTTTTGATGGAAAAGAATTTTGGGCCCCTTGCATAGAAACT
TTGGTACAGCCACCGTAAAATGGTGAGATAACACCTGCATGTACTG
CAGAGGCGCATGGAAACGAGTCACTTCTATAAGGGTAGGACACGAC
CATCGGGTCTTCTTCGCTAAACAGGGCACCTCCGCCAATTTCATAC
CCTGTATATTTGACTCTCCTATTCCCTACGGCAATGGCGGAGTATG
TCCATCCACCTCTATCACACAGTGCTGGACATCTGATCATGTATTC
TTTATTATCAAGTGGACCGCAGTTTTTAGCATTTAAACCACATTCG
TTATTGGTACCCTCCCAGTTCAAGTAGGAATTGCAACTTAAAGACA
ATATAGGAATCTTTTCTGAGCCATCATTTGGATGGAAGTATGGCGG
CTTGATAAGATAGGGGTATATGAGAGAATAAAAAATG >YNL093W Chr 14 (SEQ ID
NO: 24) CTGGCTATTTATATAAGATGTTGCTCTAATTCCATGAGTGATGTCC
CATCGCATGATGAAGAAGTTATGAATGAAGTGAACAATATCATCGA
ATTACAACAACGGCCCATGCAAATGACCAAGTCTACCGTACGTACC
AGGAGAAGACCACCACCACCTCCCATTCCTTCCACTCAAAAGCCAT
CGTTGACAGAAGAGCAAACCGAAAGTATAAGAATGTCACGCCGTAA
TAAAGATGAGAACAATGCTAAAAGAGTTGCCCCACCACCGTTGCCG
AACAGACAGTTACCTAACTTAGACGCTAATACTTACTATGTACCAT
CCTCACAAAATGACTATGGCATGTATGGTGCTTTCATGGATAAAAA
AGCAGATGAATGGAAGAGAAGAGTAATGGACTCAATTCAAAAATTG
AGCAATCAAGACACCACATTGATGTTCTTTTCGGATCCGGCATTAA
GTTTGGAAGACAGTATTCGCAGAATTAGGGAGAAGTATTCAAACTA
AATACACTGTTATTTATATACATCGGGAGGGAGTTTAAAAATTGTA
TACACTAAAGTGACCTTATGAATTGGAGCCATTTTCAACGTTTTAG
AGTGATGTTGCATCCTTTTCATCAGATCGGGAAATTAAAAAAACGG
TTATAGAAGGGCTAAATGTAAATATGTAAATTTTATGAAGTCAAGA
GTAAAGAAAATCTTCGGTTGCCGATCAAATACACTCTTACAAAATA
CCCTAAACTCAGTTTATCTTTCCATTCCAACTGATTATCGCGTACG
CCTTGGTGACCAACCATACCCTAAAATGTCAATTTCCGGGGCGGAA
GGGGTCCTTAAACTGCAAAGCCTCTTCCCATGTAAATAAAATAAAT
CCGTAGAATTTATTAAATATTGTCATATTATAAGTCGGGAAAGGCC
TTCTGAATGCACGTATATTGACCAAATAGAGTTAGATTCAAATTAT
TTATTACATTTTTTCAGCATCGAAGTTAAGTAGAATG >YLR381W Chr 12 (SEQ ID
NO: 25) GGCGAAAGAGCTGTGTATGAGAATAACGAGACCGGTGTCATTAAGA
ACCTGGAATTGCAAAAGGCCACCATCCAGGGTTACGACAATGACAT
GAGGCCAGTCATCCTTGTGAGACCTAGATTACATCATTCTTCCGAC
CAAACTGAACAGGAGCTGGAAAAATTCTCCCTGTTGGTCATCGAGC
AATCAAAGCTTTTCTTCAAAGAAAACTACCCCGCATCCACCACCAT
CTTGTTCGATCTGAACGGGTTCTCCATGTCCAATATGGACTATGCC
CCCGTCAAGTTTCTAATCACTTGTTTTGAGGCTCACTACCCGGAAT
CTTTGGGCCATCTACTTATCCATAAGGCCCCTTGGATCTTTAACCC
GATTTGGAACATCATCAAGAATTGGCTGGATCCGGTCGTCGCGTCC
AAGATTGTTTTCACTAAGAATATCGACGAATTGCACAAGTTTATCC
AACCCCAATACATCCCTAGGTATCTGGGCGGAGAAAATGACAATGA
TTTGGATCATTACACTCCACCAGATGGCTCTTTGGACGTCCATCTG
AAAGACACCGAGACTCGCGCTATGATCGAAAAGGAGAGAGAAGAAT
TGGTCGAACAATTCTTGACTGTCACTGCCCAATGGATTGAACACCA
GCCATTGAACGATCCAGCATACATTCAACTGCAGGAGAAAAGAGTA
CAACTTTCCACCGCTCTTTGTGAAAACTACTCCAAGTTAGATCCGT
ACATCAGATCAAGATCCGTTTATGACTACAATGGTTCTCTAAAAGT
TTGAGCCAAGCCACGCGACGCTCCATACTGTATAGACCTTTATTAT
AAAAACATCTATTTCTTTATATGTCCCTCTATCTTATTTTTCTTTT
ACTTTTATTTGTATAATACGCGAAAAATTATAGAATCATTTATTAT
CAAAAACAATCAACTATGAAATGAAAAACAATACGCCTGCTAATAC
GGTTCGCCCTTCAATTTTTGTTTTTTTTACATGAATG >YKL159C Chr 11 reverse
complement (SEQ ID NO: 26)
AGATAAGAATATGAGGGTAACAATGAATGAGAACTCACTTTGTAAT
CACCAGTTGAAGAATGGTAAACAAGGCTAGTAATACATCCTGGTAT
TTCGGTCTCATGTTCAAAATAATCAGTGGTAATTTATATTATGTAT
ATCTTTTCCTTGTTTCTTTGTTAAGCTTTTGTGTAAACATACTTTA
GTAATTGTGACGTTTTTTGGCACTTCGCACAGGCGTGACTAGTTTC
TGTGAACAGAGGGCGTCTATTCATGATCTTCACAAATCTTGGGGGA
GTTAATTTACTCTGCTTCCACATTACATTGATGCGCAGTGATCTAC
TGACGCCTGCTGGTGGTGAGTACACATTAGATAAATACCTAGGGTG
TGCAGCAATAGACCTTAAACAAGGGTGAGCTTTGCCTATTAGAGAA
GAACCTCTAACTGCGCTTTTCAATAATAGCCACCGAACAATTGGCA
TATTATGCAACCTGTGTTTTTAAGTACCTTTTTATGTCAGATTTTT
TGAACTATCCTTCGATTGTTATTTCTTGATATAACGTTTACATGAT
TTCTTAATTTACAACGCGGACGAATTAGATTGAAATTATTTAGCAT
ATCTGTCTAAGAGCACATTAGATCGAAGTACCGTAGCACATCTGCC
ATACTATCAAATGCATCATCACGAAAAGGCGCAGTGTTGATGGATA
GTAAATATGAAAAGGATCTCTTCTTAGGTGTTCCAAGCCTCGCTTC
ACGTGTAACATACACAGTTTAGAAGTTTACCAAACCGCGGCAAGGC
GCTGTAAGGTTCAAGAGGCTTTGAATTATGCTATCTTACTATCTTG
CCGCAAGTTGCTGTCAGGTGTAGTTCCTAGGCGGCTCGTTGACAAG
ACCCTTCTGCCCAATGACTTCTATATAAAGTATGGTTTTCTTTTTA
AATACAGTAATTCCATGAAAAAAAGAGGGCCAAAAAGATCAAGCAA
TAAACCAACCGATATATAAAACACAGAACTGCAGATG >YIL058W Chr 9 (SEQ ID NO:
27) ACAGAAGTAAGTACCGGGGTACCTAAATATACGCATAAAAGTCTCT
TTCTTTTTTTTTTTTTTTTTTTTAGCTTCCTACATTTCGTTAATAA
TATTATATAGATTATATTTATATTTAAGAAAAGTAATATCAGCATA
TTATGAATAGACAAAAAAGTCTAAGGTCAAGATTTATTAAATGTTA
GATTATTAAGATTACAATTAACAACATTTGTTGTTACTTCATAAAA
TTTCGTTACTTCATAAAAGAACTACAGTCCCCCACTTCTAAATGAG
TGGTTGTAGGGGTTCATATACCTCCCATACTGTTGGAATAAAAATC
AACTATCATCTACTAACTAGTATTTACGTTACTAGTATATTATCAT
ATACGGTGTTAGAAGATGACGCAAATGATGAGAAATAGTCATCTAA
ATTAGTGGAAGCTGAAACGCAAGGATTGATAATGTAATAGGATCAA
TGAATATTAACATATAAAATGATGATAATAATATTTATAGAATTGT
GTAGAATTGCAGATTCCCTTTTATGGATTCCTAAATCCTCGAGGAG
AACTTCTAGTACATTCTACATACCTAATATTATTGCCTTATTAAAA
ATGGAATCCCAACAATTATCTCAAAATTCACCAACTCTTCACATAC
ATACCTGCGGTAGCAAGATAGGTACACTTTTTCTACGTTTCACGAA
GGTAGCGATAGGTACCTCACTCATTGTAGGTGCGGGGGTAGCGATG
GAGGTTTCTGTACCATTACCACCACAGCCACTATACTCACGTAGTG
AAGTCCCTAGCGTGGAATTGTGTGGTATCGTTGCTATCTGTCGTTC
GCCACCCTCGGTATATCCAACGTGCAGGCCAATATCACTCTCTAAG
AAAATCGTATCAGGATTGGTACGGACAAACTCTTCATAGCCTACGT
CGAAATCAAATTCTGCTAAAGCTGTGACTGTAGTTAAACCTAAAAC
CGCGATTATTGCTTGAAAAACTGTGGAGAAATTCATG >YKR037C Chr 11 reverse
complement (SEQ ID NO: 28)
TGTCATCGCCGCCAGAACGTGCTCCTTACTGTGGGACGTGCCACTG
GTGGGAGTAAACCACTGCATTGGTCACATCGAAATGGGGAGAGAAA
TCACTAAAGCTCAAAATCCTGTGGTACTGTATGTAAGTGGTGGAAA
TACACAAGTTATTGCATACTCGGAAAAAAGGTACCGTATCTTTGGT
GAAACGCTTGATATTGCTATCGGTAATTGTCTTGATAGATTTGCAA
GAACTCTGAAGATACCTAATGAGCCCTCGCCTGGCTACAACATCGA
GCAGTTAGCTAAAAAAGCCCCTCACAAAGAAAACTTGGTAGAACTT
CCCTATACAGTAAAGGGGATGGATCTTTCGATGAGTGGTATATTGG
CTTCCATCGATTTACTTGCCAAGGATCTATTTAAGGGCAATAAGAA
AAATAAGATCCTATTCGACAAGACAACGGGCGAGCAAAAAGTCACT
GTAGAGGATCTTTGCTACTCTCTGCAAGAGAACCTATTCGCCATGC
TAGTTGAAATAACAGAAAGAGCTATGGCTCACGTTAACTCCAATCA
AGTTTTGATCGTAGGCGGTGTTGGTTGTAACGTGCGATTACAAGAA
ATGATGGCGCAAATGTGTAAAGACAGGGCCAATGGGCAAGTACATG
CTACAGATAATAGGTTTTGTATCGATAACGGAGTTATGATTGCCCA
AGCAGGACTACTAGAGTATAGAATGGGTGGGATCGTGAAGGACTTT
TCTGAAACTGTTGTTACGCAGAAATTCAGAACCGATGAAGTATACG
CAGCCTGGCGTGATTAACGATTTTTGTTACAGTTAGCCTAATGTTC
ACACTACGAATATATATATATATTTATATATGCCATACACCTTACA
TTTACAGCTCAGTGTATAACCTTGTGGAAGCGCGAAAATAAGCAAG
AGACATTTGAAAATAATCAATAAACAAAGCTCAAAAAAATTAGATC
ATCACCTAACAAAATAAGACGTGGCAATTACAGAATG >YNL279W Chr 14 (SEQ ID
NO: 29) GGCAACATAGCAGTACGTCGCCAAAAAGAACGAAAACAAAATAGAT
ATTATGCATAAACTAACGTGATTTTCATACAGATATTGCAATTCAG
GCAATTGTCCATCAGTCAATTTCCATCTGATAGCCAAAACTAAGAC
CAAAGTTGTAGACATGGCAATTCCATTGATCTTATACGAAAGCTTC
GAACCATCCCTTAACTGAACACCCTTCATGACTCTGCCCGGTAAAA
TGACGTCCAAAACTGCCAGTATTCCATACCACAGGCAATAGACAGT
CCATAATTCACGATTGCCCAGATAGTAGCGCAATGGCTTGATACCG
TTCCAAAGCTCAACTATATCGAAATTCTGGAAAAATCCCTTAATAA
AATAATCGGGCCTTATCATTTGATTCAAGATGATAGTGAAAACAGG
CAGCCCTATGCTGATGCCTAAGGCACCAATCAGCCCACCAAATTCA
AACTCTGTAGTTCTGGGATTCAAAGCTGATACCATCCTTTTATTCT
CTACACCGAATTTGTCTTTACTCCTATGCTGTTTACAAGGTCTATC
TGATAAGCAATTGCGCAAGAAAATAGTAGAATGAAAACTGATTATT
AAAAACAAACGTAAACTCAAGCCTCACTTGATGCTCAGACGGAGTA
CGTGAAAAACGTCCGTTATGCAAAACCCTTTATATGCACAACCTTC
ACACAATGCAAATTTCCGATGATGCCTACATACAAAAGAGCGAAAG
GCGATATAAATTTTTTTCACGGGATTTTCGTTTAGGTGAAAATAAA
ATGAACGACAGAGCATGCAGAGTCCGGGTAATACATATGTTTCAAT
ACTGTTTCAATACTGTTTCAGAAGTGCGTCACATATTAATTTTAAC
TTATAACTGGCCTGTTGCTGGCAAGAGGTATATATATATGACGAAT
GTGACCAACATAAGTCCTTAAGATAATCCCGAAATATTTGGTTAGG
ATGATTCCCTTTCGAATTTGTGAACGTTGATGATATG >YOR032C Chr 15 reverse
complement (SEQ ID NO: 30)
CTAGCGGTCTACATTCGCTATCAACTGAACGAATAGTGATGCAATC
TGAACCTTAAACTTTTCGCTCGCTTTTACTTTCCAGAAAAAATTTT
CTTCATTGGGATTCCTCTGCTGACAAACGGGCTCACAGAACTGAGG
AAACGAGAATTTTGTTAATCAAAGCAACATTACGGGAAGGAACGGA
ACCGTCTCGTTGCTGTGGGCGCGAGTTCGACGCAAGAAAAAAATGC
TGCAGCATCGTCTTTTGTGAACGCCAAGAAGGAGTCCGGCGAAGGT
ATTGGGCGTCACAACCTTTTATTCTCCGCAGTGTCTCTTGCCTACA
AAGCCTGCAGAAGCTTTTATGCCGGGGTGTACGTATGTTAATCAGT
TACCCTTTCCTACTGTGTGCCGCACCCCTCATTTGCTCTCGATACG
AGAAGTTCCGCGCCGATTCTGGCCTTTTTGTTTTCTGGCTAGCCTA
CTTTCAGGGGACGTGCCTTCTTATGCCGCTGTAAATTACAGCAGAG
ATTTAGATCTGAGTGAAAACGTGGCGAGGAAAAAGGGTACACCGTT
AAAAGCGTAGACACAATCCCACTTTGTTGTGCCTAGTCTTCTCTGC
TGTATTTTAATTTTCTGTTTGAACTTTTTTCCGTTCTTTTCCATCA
TTGTTTAATTTTTACTTTGAGTGGGCTTCTCAAAAGAAAAAAATAG
CTGGTATTGTATTTTTACCGTAGGCGTTTTCTTTGTATTTTTCCCA
AGAGCAGCAGCACCTTTTATATACCTTGGTTACACATTGTAAGCCT
CAAAAATCGTCCTATGTAAGTGTCTTATCGCATCATACTTAAGGTC
AAACCTGTAAAGCGCGCTTCTAGTTTTATTTCTACTACTCGCCTTG
GCTCTCGAGATCGCTTAGGTAGTTTAATAGTTAGTCACACCCACCC
ACCCATTCGCTTAGAATATAGAATATAGAGCAGTTGCAGTACACGA
CTCATTTTAGCAACGATAAGAGTATAAATTGGAAATG >YML058W Chr 13 (SEQ ID
NO: 31)
TCCTTATAACCTGTAAAAAAAAAAAAAAAAAAAAAAAAGAAGGGTT
TAAATAAAAATCGGACTTACTCAAAGGGTTGAAAAGCACTTTAATA
TAGGTTTTTAGTTTCGGGTAAGAAGATGTGTCAAAGGTCTCGAAAA
GGAAACATTAGGGCAAATACGTAAAGGTTGCCTTCGTATCTCGTTT
CCACAGCAACGAGAAACGAGATACGAAGGCAACTGATTCCCAGCAT
ATAAAAGGCCATGAATTTTTGGCGTTTCCTATTGTCTTTTCGAAAC
TATTTCTTTCATTCCTTTCCTTTCTTGTTCTTTTACTTATAAGAAA
AATAATATACGTATATATATAACTACAAACCACATCAGCAATAAAA
AAAAACTATATGACCATGGACCAAGGCCTTAACCCAAAGCAATTCT
TCCTTGACGATGTCGTCCTACAAGACACTTTGTGCTCAATGAGCAA
CCGTGTCAACAAGAGTGTCAAGACCGGCTACTTATTCCCCAAGGAT
CACGTTCCTTCTGCCAACATCATTGCCGTCGAACGTCGCGGCGGTC
TTTCTGACATTGGTAAGAATACTTCCAACTAAGAGCATGCTTCTCT
TTTTTTTTGTAGGCCAATGATAGGAAAGAACAATAGATTATAAATA
CGTCAGAATATAGTAGATATGTTTTTATGTTTAGACCTCGTACATA
GGAATAATTGACGTTTTTTTTTGGCCAACATTTGAAATTTTTTTTT
GTTACCTCGCGCTGAGCCCAAACGGGCTCCACTACCCGCCGCGGTC
GCCATTTTGGGAAGTCATCCGTCCCAAAAAGGAAATAGCCATAACA
TATCGTTACTGTTTTGGAACATCGCCCGTTTCGCCCGATTCCGCCT
CAGCGGGTATAAAAAGAGATCTTTTTTTTTCCTGGCTGTCCCTTCC
CATTTTTAAATGTCTTATCTGCTCCTTTGTGATCTTACGGTCTCAC
TAACCTCTCTTCAACTGCTCAATAATTTCCCGCTATG >YMR303C Chr 13 reverse
complement (SEQ ID NO: 32)
AATGGCAAACTGAGCACAACAATACCAGTCCGGATCAACTGGCACC
ATCTCTCCCGTAGTCTCATCTAATTTTTCTTCCGGATGAGGTTCCA
GATATACCGCAACACCTTTATTATGGTTTCCCTGAGGGAATAATAG
AATGTCCCATTCGAAATCACCAATTCTAAACCTGGGCGAATTGTAT
TTCGGGTTTGTTAACTCGTTCCAGTCAGGAATGTTCCACGTGAAGC
TATCTTCCAGCAAAGTCTCCACTTCTTCATCAAATTGTGGGAGAAT
ACTCCCAATGCTCTTATCTATGGGACTTCCGGGAAACACAGTACCG
ATACTTCCCAATTCGTCTTCAGAGCTCATTGTTTGTTTGAAGAGAC
TAATCAAAGAATCGTTTTCTCAAAAAAATTAATATCTTAACTGATA
GTTTGATCAAAGGGGCAAAACGTAGGGGCAAACAAACGGAAAAATC
GTTTCTCAAATTTTCTGATGCCAAGAACTCTAACCAGTCTTATCTA
AAAATTGCCTTATGATCCGTCTCTCCGGTTACAGCCTGTGTAACTG
ATTAATCCTGCCTTTCTAATCACCATTCTAATGTTTTAATTAAGGG
ATTTTGTCTTCATTAACGGCTTTCGCTCATAAAAATGTTATGACGT
TTTGCCCGCAGGCGGGAAACCATCCACTTCACGAGACTGATCTCCT
CTGCCGGAACACCGGGCATCTCCAACTTATAAGTTGGAGAAATAAG
AGAATTTCAGATTGAGAGAATGAAAAAAAAAAAAAAAAAAAAGGCA
GAGGAGAGCATAGAAATGGGGTTCACTTTTTGGTAAAGCTATAGCA
TGCCTATCACATATAAATAGAGTGCCAGTAGCGACTTTTTTCACAC
TCGAAATACTCTTACTACTGCTCTCTTGTTGTTTTTATCACTTCTT
GTTTCTTCTTGGTAAATAGAATATCAAGCTACAAAAAGCATACAAT
CAACTATCAACTATTAACTATATCGTAATACACAATG >YJL153C Chr 10 reverse
complement (SEQ ID NO: 33)
TTCTTTTTTTTTTTTTTTTTTTCTTTTTGGGAACGGGGGAAAAGGT
CTCACTAGTATATAAAGGAAAGAGAGTACAGCACGAATGAGCCAGC
TGCAGGAAAGGGGCTAGGTTAAAAAATAAAAACCCAGCAGAAAGCA
AACAACCAAATATAATTTAGAAATGGACAGAGACCATATTAATGAC
CATGACCATCGAATGAGCTATTCCATCAACAAGGACGACTTGTTGT
TAATGGTTTTGGCGGTTTTCATTCCCCCAGTGGCCGTCTGGAAGCG
TAAGGGTATGTTCAACAGGGATACACTATTGAACTTACTTCTCTTC
CTACTGTTATTCTTCCCAGCAATCATTCACGCTTGCTACGTTGTAT
ATGAAACGAGTAGTGAACGTTCGTACGATCTTTCACGCAGACATGC
GACTGCGCCCGCCGTAGACCGTGACCTGGAAGCTCACCCTGCAGAG
GAATCTCAAGCACAGCCTCCAGCATATGATGAAGACGATGAGGCCG
GTGCCGATGTGCCCTTGATGGACAACAAACAACAGCTCTCTTCCGG
CCGTACTTAGTGATCGGAACGAGCTCTTTATCACCGTAGTTCTAAA
TAACACATAGAGTAAATTATTGCCTTTTTCTTCGTTCCTTTTGTTC
TTCACGTCCTTTTTATGAAATACGTGCCGGTGTTCCGGGGTTGGAT
GCGGAATCGAAAGTGTTGAATGTGAAATATGCGGAGGCCAAGTATG
CGCTTCGGCGGCTAAATGCGGCATGTGAAAAGTATTGTCTATTTTA
TCTTCATCCTTCTTTCCCAGAATATTGAACTTATTTAATTCACATG
GAGCAGAGAAAGCGCACCTCTGCGTTGGCGGCAATGTTAATTTGAG
ACGTATATAAATTGGAGCTTTCGTCACCTTTTTTTGGCTTGTTCTG
TTGTCGGGTTCCTAATGTTAGTTTTATCCTTGATTTATTCTGTTTC
ATTCCCTTTTTTTTCCAGTGAAAAAGAAGTAACAATG >YLR092W Chr 12 (SEQ ID
NO: 34) CGCTCCCACGACATGCGCATCTCTTCCAAAAACACTACCACTTTCT
ACTGACTCACTGGCCATTTGAGAACCACAGAGACTTGGTAGATTCC
ATCGCTGTAAACAATGGGAAACTAAATTCAACGTCTTCAAGGAGCG
TCTGGCTGAAGGCGGATTGGATAACACTGTTTAACGTTAAAAACCC
TTGGGTCCAAACGCCGCCATCGTTAATGCGTCTGAGTGGGACAGAT
CTTGATACCTTCACACCAGAGAGGATATTCCTGATAAATTCTCTTG
GAAACCACTACAAATTTTTAATAGCGAACAGTCATCTAAGCTACAA
TCACAAAAAATACCCCTCCCCTGGCGTGCAAATTCCTGTCAGGAAC
GCTCTTGGCGAAGTTTCTCCGGCCAAACAAATTGCCCAACTTTTCG
CAAGACAGCTGTCTCATATTTACAAGAGCCTCTTCATAGAAAACCC
CCCGCTCTCTCCTGAGAACGAGCTGGCATTGACTGCCGTCTTCTAT
GACGAGACTGTAGAGCGACGCCTTAGAAGGCTCTATATGCGAGCTT
GTGCAAGGGCATATACGACTACTAATGCCGACTCCACCACGGAGCC
TTTGATGTTTCATTGCACCCGGTGGGAAGTTGATTAGTTGTTGTTG
GCTGTTATACTGGAGTAAATGTCACGTGCCACTGTGGCCGTGTCAG
GTCACGTGAAGAGAGAAATTCGGCCACAGTTGTGGTCCTAAACAAG
GACTGAAAAAGATCGCAGAAACTAGCAACGTCGGATAGCGGTATGG
TTGAACCAAAGTCCAGTTTTTAGTCATAGCCGGTTGAGGGATGATC
TGCCGTTCAGTGTTCTGCGTTCAATAATACGCATATATATATATAT
CAAGGTATCAACACAATACTTGAACAGGTTAGGCTCGTACTAAAGC
CTGGAAGTTTTTATCTTTCTGTAACATTTTTCTTTCTTGAGGTGTG
TGTGTGTATAGATTAGCAGGGAATTATCTAAGATATG >YFL052W Chr 6 (SEQ ID NO:
35) CGACTAACCTTTTTCTTGGCAGTATGGAGACTGACTAGGTCTCCCA
AACATTCATTGTAACTGCTGTTTAAAGATTTTGTTCTAACCTAAAT
TCAAGTGAGAAGCTGAACATGTGTCTCTACTTATGATATCACGACA
GCAAATACTAATCTTGCCATAAATAGTCTAGCGTTTTGCAACTTAC
CTCTAGATATATTTTATTTCTTGAGGAACCGTTTTCGTCGGTAATA
ACAAAATACTACTGAAACGCCACAGCATTGAGAGAATACGTTATCG
ATTACGGCTTTCTTCTCGCTCCAGATGTCGCGGGTAAGATATTCAC
CTCAAACTTTTCTTGTTGAGTGTCGTCACAAATCTAGAACCTACAT
GCCATCTCAACGATTTTTCTGGAGAAAGGCCTCACTCCGTTCCGTA
CGTAATGCATAGATAAAGTATCAGGATCTTCACGATGCTCGAGAGT
TACTTAGTAGTCTGAGTTTATGCGAAAAAAACTCCGCCGTTGTAAT
AATCGGGAATACACAGAAGTAGTACTGCACTATCACTGGGATACTC
AAAAACCTTCTTTTTAACTTTTCTATCCCACAAATAGAACATAGGA
AAGAACATTGACTCCTCCACTTGAAGTTAAATTACAGGAACAAACG
CCTAACTATAATTTCGACATTGTTGCATCAACGAATCGACCGAAAG
AAAAATCTGGAGTTGCAGTTATCACTTGTATGTGCACTAAGATTTA
TATTTTTACTCCTGAGATCTGCCAAATCGGTAGCTTATTGAACTGC
GTTCCTTTTTCCCCTGAGTTCTCGAGGTACCTGCGGCTTTGTCTGT
GCCATCTCCCCCACTTTAAAGTACCCCACGTTACTACCGCGTTTTT
CCCCACCCCCGGCTTAATAAATTAGCTATATCTTGTTGACTTAAAT
ACGGAGAAAAGAAGAAAACCTTCAAGAAATGCTTCATTGTCTTGTC
AAAAGAGCTAAAGTAAAAGAGCTCTAGTTCGAAGATG >YIR017C Chr 9 reverse
complement (SEQ ID NO: 36)
CTGTCGAGTGCTGGTGGCGCTTGATAGCGGTATTGTACTTAATAGG
TATCGCCTTCATTGGTTTGAAATTCTCGATCAAATTCTGCAGGATA
GGGTCCTGAGCAGGCACAGAGGCTTGTTTCAGAGTAAGGGTCTCCA
ATTTCTGTTTAAGGGCATGATTTTCCGATTCTTTCGCTTGAAGTTC
ATTTTGGAGGCGGTTTAGTTTCGTTTCGTAGTTTTTAACGACCTTA
CTCAGTGACTCTATGGTTTCCTCCAGCACTTGCAACTTGTTGTTTT
TGCGTTCTCTGTAGGCCCTTTGGGCGTCTCTATTCTGCCGTTTTTT
CTTCTGTAGCTCCTCATCATTTTCTTCAGTTTCGTAATCTTCGTGA
AGTCGATGAACTCTTTTCCTCCGCTGAGCTGCTCTATGGGGCAACC
TTGGAGGCAGTTTCCAATTTTTGGACACGTGGATCTTGGACAAAAG
AGCGAGGTGGCTTTCCTCAGACTCCTTAGGTTTTATCAGAGGTAGA
GCCATGACTGTGATAATATGCTAGTTACACTGTTTATGTTGTGTGA
ACTTGTTGTAATATGGTTAACTTCACTTTCAGTGATTGATATGATA
GCGACATCACTGCCGTGCAAAAAGACCATTCCATTACTGCACCTTT
TTGTCCTTTTCCGTGGAATAAAAGTTCACTCGTCAGTTCCATGCAT
TCTGGAAAAAAATGATCTGAAAGATGCCACAGTTGTGGGGCCCGCC
CGGCCCAATAGGTAAACTAAAATACAATAGAAGGGGTACTGAGTGC
ACGTGACTTATTTTTTTTTTTTGGTTTTAGGTTTCGCTTTTTTCAC
CTTTTTCTACTTTCTAACACCACAGTTTTGGGCGGGAAGCGGAAAC
GCCATAGTTGTAGGTCACTGGCGTGAGTCAAGGCCGGGCAGCCAAT
GACTAAGAACACGAGGTAACTTGAATTTAACTATTTATAACCAGTG
GTAGTTACGAAGACAAATTGTTTTGTTCGTCAATATG >YLL062C Chr 12 reverse
complement (SEQ ID NO: 37)
CAATTTTGAAAATTGACATATAAAACTCTGATTCTGCAAAGCCTCT
TGCCCCAAAAAGATTCAATAGCATGCTTAAAACGAACGCGATTGCA
ACCCAGGCAGCAGGATTGACCGATGGTGCCCAATAAGAGATGGTAA
GCGAACATCCTACCAGCTCGCTAGGGTAAGATATGAGCCACATCAA
TAAGTAGTTAATTCCTATACTGAAGCCGACTGATGGGTCTATAAAT
CTACTGGCGTGCAGCGCATAGGAGCCGGAAACAGGGTACTGACATG
AAAGTTCGGCCCCACACTGAACAACACAGAGCATAGATGCGCCGAC
CAACAAGAAACCAATTAGCAAAGAAGCAGGTCCGGACGCTAGTGAT
TCCCCTAAACCAATGAAAAGACCCGTCCCTAGAGTACCTCCAATGG
CTATCATGGTCAGGTGTCTTTGATCTAGGACCTTTTTATACGGCTG
GTTGGCTAAGTCCCATTGTATTTTTTCTTCCTCTGAAAGCTCCGAA
TACAGCAAACCTTCTTGGATCGCTGGTGAATCATTTCGCTTGAAAG
AGTTCTTGAATCTGTGTATTAGATTATTTCCTTCATTCGAAGGCGT
ACTTAACACGCTAGTGGAAAACTGAACCTTCGAAAGTTTGGTAGAT
TCAAATTCATCCATTTCGGGTATCTTTCATGGTATCCTGATGGAGA
ACTCTTGACGGTGATTACTTAACCACCCAAATCTGTAAAATATATA
CGCATTTATTTCCTGGCATCTCTTCGCTATCCCCGCAAAAAGTTAA
TGAAAAAAAGCCATTCCTGCAGGAACTGTGGAAAAGTAAAAACTGT
GGCATGAAAGTACTACCAAATTGTGGCGAGGTATTCAATGGTAATA
GCAAAACAGCAACAACGCCAATTTGAAAATGTACTACTAAGGAAGA
TAATTTAAACAGACAAGTTGTAGAAGAAATCTGAATCAGCAACTTT
TAGTGTGCAACTTGAAAAAGTAATCAAAAAACAGATG >YMR323W Chr 13 (SEQ ID
NO: 38) CAGTTTTCTTTTTGAATGAAGGAGAAAAGTGTTCTAGAAATATGTT
TTAGTTCTAAGTATCGTTGTCTGGAGCAAGCAAAGAAAAGCAGACC
TGTATGTGCAACTTTCTTGTTTTCCATTAAAATCTCAAGAACCGGC
CCCTTGACTGTGCTCATATATCAAGTATCGGTTTTAAAGTCCCCTA
ATTGTGATAATTGCAAGAATGACAGTCAAAAAGATATTTTTCTTTT
CTTTCTTTTTACCTATCCTTCATGAAAATACCTATGCTGATACTGA
ATATCTTCATTTGCAGCTATGCAATATTTCAGATACTGCATTTTAC
CCAAGAGTTTCTTTACTGGTGGTTGGATTTTTTTTAAGACGAGAAG
ATTACCAGTCCTGGATGTTCATGAATGTGCCTATTGCGTCATCAGC
GACTCCGCGCCAAGAAAAGAAATACTCGCATGCTCGGATTTATAGA
ATCGCATAATGGAAATAGCTTCCTTAAAGGATGGAGATAAGGAAAA
AATGACAAGAACAAGGAAGTACTTGGGAGCTTTCTCCGACATGTTA
AATATTTGAAACTCACGCAGCTCTCCTGGAAAGTTGCATCTGGAAA
GGTAAGGTTGTTTTTTGTTCAGACATCAAAATCGGCTCTTGTAGAC
AATGTCACAGATCCACAAGTTGAAAAAAAAGTTTCGATGAACTGGA
TAAGGGGAGAGGTCACAAACAAAGTGTAGGGGGTGAGTAGTATAGT
GGTAACTTGTTCATTTACCTAAATAACATTCTCACTAGAAAAGAGA
TATTGAAAGACTCCTCGTTTACCTAACTTGGCTGGTTCTTAGGTAT
ATAATAGAAACATGGATCTGGCAGGTTATTACTTACAGTTTGCATT
CCCACCAGGTAACAATTTTCATTATATATAGTTATTTTTTACCACC
ACTTCTTCTTCTTTCACTTTGTCTTGCAATAGAAATACCAAAACAA
GGTGAGGAAAAAATCGAATCTCAACGATAAAAATATG >YPL171C Chr 16 reverse
complement (SEQ ID NO: 39)
CGGTTGGATCCTCACTTGTTTTAACACCTCCAAATAACAAGTTTTT
AATGAAGGACATTTGTTCTCTATAATATTCCGATGTACGTGTGTGT
GGCTGATGAGATTTAGACTGGTTAGACTATTTGACGCGTCTATTAT
AGCTTACTGCAACAAGAAAATGATCGTTGATATATAAACTCTCAGA
TGTATATATCGTTCTGGAAACATCGAGCATAATACAATACAATTCA
ACAAAAATGCGAGAAGGCACTGATGTCTTGTCGTTAAAGAACCAAA
AACGCGGACACTACGACCGTCTTATTTCCGGTAGAAAAAGGGTACA
TACAGTTGAAGGAACGAAGAAAATTAAAATTAGAAAAAAAAGTAAA
ATAAAACAAGGAAGGTAGGGTAATATGGTCTCGTTTCCTTTGTCGC
TCCGCAAATAAAGGAGCTTATTCCCGCACGCTCACATGGTAATTTG
CGCCAAATCACGGATGTGGAAAACTGATCACGTGCTTCGATCGCCA
ACTACTGAGCGTCGTCCCACACTGATCTGGCACAGCTTACCTCGCC
TTGAAAATTTTAATCTGTCCTGCTCGTTTGTTGTATATTGCTTCTT
CTCAGAATATGCCCGCGATAACTGACAAAGAGGGTTCGACGTTTCA
GAGATTCTACTCTTGACCACTGTTTCGTGTAGCCGCTCAAGGTTTA
TTTCTTTCTTCTTTAATGTTCTTGGCACTTAGGCGGCTCCGTCCTC
CGTCTGAAATTGCCGATCCTATTATTTGCGGAGGGCTCCTTAGAAG
GGCTCCTTAGTAAGCAGTTTGCGTTCCTGATATAACTCCGTTCAGA
ACAAGGATAAAGTCGCAATAACCATTACTAAGCACAGTGTTGTAAG
TAGGACAACTCGAACCTATATAAGGGTTGTGAACTGTGCTTGATTC
TTGCCCATCATATGCAAAAAAGTACGTACTTGATATATACAACAAC
TGTAGTTCAGTATAGCGAAGTTTAAATTTAGAAGATG >YER065C Chr 5 reverse
complement (SEQ ID NO: 40)
TGGAAATGTAAAGGATAATGAGTGAGCATATAAAATGGAAGAAAAA
ATAATAATAGGATTATGTATAAAATATCGATTCCCTTTTGTAGATT
TCGAGATCTTCGAGGAGAACTTCTAGCACGTTGTTTACATATCTAA
TGTTGTAACACGGCCTTTGTTATCTTGTGAATATATCCATCACCTA
CGTCGCTTGAGGTTTCGTATCAAAGAAAGAGAGTTCAAACATAATC
ATACGTGTTTCACTCCAAATTTGATCACAAATGGCCACCCATTATA
GCTTGTTATTATTTACTTCTTCTTGAGTTCATTTTATTGAAAGATG
CCCGCTGGGAACAAAACATAAATAAAATTCAAGAGTACAATATAGC
GTAGCATAGTTGCGAGCAAGGGAAGGCAACACAGTTAAAATGACAA
TGTTAGAAGGAAAAACTAAATCAAGCTCCCATAGGAAATTTCTTGT
GTCAGTAATATGTCTCGAGAAAATAATAATAATTGAATCTATTTAT
TGAAAAGTAAATATCTCGTAACCCGGATGCTTTGGGCGGTCGGGTT
TTGCTACTCGTCATCCGATGAGAAAAACTGTTCCCTTTTGCCCCAG
GTTTCCATTCATCCGAGCGATCACTTATCTGACTTCGTCACTTTTT
CATTTCATCCGAAACAATCAAAACTGAAGCCAATCACCACAAAATT
AACACTCAACGTCATCTTTCACTACCCTTTACAGAAGAAAATATCC
ATAGTCCGGACTAGCATCCCAGTATGTGACTCAATATTGGTGCAAA
AGAGAAAAGCATAAGTCAGTCCAAAGTCCGCCCTTAACCAGGCACA
TCGGAATTCACAAAACGTTTCTTTATTATATAAAGGAGCTGCTTCA
CTGGCAAAATTCTTATTATTTGTCTTGGCTTGCTAATTTCATCTTA
TCCTTTTTTTCTTTTCACACCCAAATACCTAACAATTGAGAGAAAA
CTCTTAGCATAACATAACAAAAAGTCAACGAAAAATG >YDR114C Chr 4 reverse
complement (SEQ ID NO: 41)
TTCCTAAGCCTTTGAGATATATCAAAATTGCTGCCTTTACCAACGA
CGAAAGTAAATTGGAAGAGCTTCGTGAGAAATACCCAAATCATTTG
ATTGGCGGAGCAGATCTTGTTGCCAAAATTAAAAGCGGTGAAATTA
GCGTTGATTTTGACAAAGCATTTGCAACTCCTGATATTGTTCCAGC
TTTACAATCGCAGGTGGCCAGGATACTTGGTCCTCGTGGGGTTTTA
CCCTCTGTCAAAAAAGGTACGGTGAGTGATGATATAAGCTCCTTAC
TTCAAGAAAGTCTAGGATCTATGCCTTTCAGGCAAAGGGGAAACTC
TATCAGCATTGGAGTCGGCAAATGCTACTTCACTGATCGAGAAATT
TTACAAAACATTATATCCGCTAGAGCGGCATTTAAAACTGCTGTTG
ATAATCAAAAATCTAAGAAACCAAATATACTGAGTAAAACCACATT
ATCAAGCACACATGGCCCCGGTATTGTAATCGATTTTGCTTAATCA
GCGAATCAGTTTTGTCTTGTTGGACTTTTTTAATTGTATATATATA
TATACTGGTGCATTTTCTTTCTTTTTATATAAAATCATGTACATAC
AAAGTAACCGTCTTTTTAAAGTGGAATAAAGTAACTGCTCTTTTAT
TTTTGAATAATAGTGAAAGTGTATGTTGCATAATCATTAGATACCT
ATGTACAAGAAGTATTCAAATTTATCTGAATAGTATGTGAAATTTC
AATGAGACAAAAACCACCTACCTTTGAGCTTCCTCCTTTTCAATAT
CTTAGAGCCCTGTTTGCTCTTGGCCCTAGCCAAGAAACCAAAAGTT
CTCTTCCGTTTCAATGTACTAGGTTGATAGGTGTTACCCCTTGATT
TCCATCTTCTTTGACCTATTAAGCCAAATCCCAACGGTGTGAAGGA
CGGAGTCTTCAATGGTGAACTGTTTAAGAGCATACCGGTTTGTGGG
CGTAACACAGATAGAGCAGAGAAAGAAGATATAGATG YBL065W Chr 2 (SEQ ID NO: 68)
CGGTCCAGAAAAGCAACTGAGATTGGGAGTATAATTCGGTGGCGTT
ATTGGAATACATAATAGGAAAATACGGCAGATACCTAGTCACGAAA
ATATGGTGTAATCTATTCGCTTTTTCAATGGAAATACTAATATCGC
CCAGTACAAACTCATCTACATTTGCATGCGGCGAAGGCAATAATGG
CATAGTCGTGGTGGTGGCTACTACCGGCGTCTTCGTCCTATCTGCA
TTGTTATTTGTTGCTACGTGGCTTGTTGCTGCAAATGGTGGTTGAT
TTGTTGTCGTTGTAACGGTGGTCGACGTAGTATGAGGGGAATATGT
TTTTTGAATTGGAGAGAAGGGGCCGTTCGGAGTGTTACCTGCAGAG
TTGTTCTTATAAAAAGCCATTTGCAAAGCAGGAGGTAAAGATTCTT
TTGAATTATTAGGCAATGCTGAGTCATTTAGCAGCGGGGGCAGCTT
ATTTGGCTCTGTTGCAACCAAGCCTTTCGAATCTTGCGCTAAAGAA
GATGCGCGCAACGTCATGTGAGAAGATGCTTCATTATTTGCTTTAA
ATTTATTCGTATTGCTGCCCTGATTTGCTTGTAAGAGTTGGGGTTC
CCTGGACAAATAAGTTTGAACTGAAACCTTAGAATCTCGTTGAGCC
GCGGAAGAAGTTGGAGAACCTGAGGAAGGAGAAGAATCTGGGTTAG
GGATAATAGTACCCGGAGTTGGAGTTGGATGCAGATTGAGCTTATT
CAAAAGGCTATTGCCCATGGGAATCTGTTGTAAAAGATGAACGAAA
ACGCTGTCATTGGCCAGAAGAGTATCGAGTTTAGATTTGATTTCAT
CCACATCTTGTCTCAGTAGTTGCAACTGTGAGCCCTTCTTAGGCCT
GAATTGAGGATTGATTTCACAGTGGAGACCAATTTTTTCGCATCTG
GAGCAAGGATGAGGGAAATTTTGACTAGCATCGCATTTGATTTTGT
GCTGTCTACAATGTGTACATG
TABLE-US-00004 TABLE 3 Baseline promoters identified Affymetrix S.
cerevisiae Probe ID Gene Synonym Proposed function 4464_AT YHR082C
KSP1 Serine/threonine protein kinases 6604_AT YDL126C CDC48
Microsomal protein of 7242_AT YBR106W PHO88 CDC48VPAS1VSEC18 family
membrane protein involved in inorganic phosphate transport 10388_AT
YLL043W FPS1 Suppressor of tps1Vfdp1 7639_AT YPR108W RPN7
Regulatory particle of the proteasome 4130_AT YIL041W 11172_AT
YJL166W QCR8 Ubiquinol cytochrome-c reductase 6311_AT YDR165W
9999_AT YLR330W 6540_AT YDL058W USO1 Integrin analogue gene 4531_AT
YHR012W VPS29 10644_AT YKL053W 10766_AT YKL196C YKT6 v-SNARE
6583_AT YDL103C QRI1 9874_AT YLR427W 10368_S_AT YLL020C
[0104] Baseline promoters identified in the experiments set forth
above are as follows:
TABLE-US-00005 >YHR082C Chr 8 reverse complement (SEQ ID NO: 52)
TCTAAATATCGTCCCGTTACCCTCATCGCTACTGCCGGGCGTTCCC
AACCCCTTCCGAGGCAGATTATATTTATTTATCTTTTTAACAATTT
GTCCCTATATAGGCGGGCCGAACACAGCACGTGAAAGAACTCTCAG
GCTTCCACAACAGGCCATATCAAGTCCACACTTCCCTCATTACTAG
CATTTTTACACAAGCAGTCCACGCATTCCGCAACAGTGTTCACCTT
TCTTTTCCCTAACTTATCTTATTATTTGTTTGTTGATCGCAAACAG
ACCCTTTTTTTTGGTGAGCGCATTTTACACAGTCTCAAGTGTCCTC
TACTATAAAAATACAACCTCAAACAGTACTTACTGGCGTCCGCAGA
ATATTACTGCACTCTCAATCTGTAAAGAATATAACAAGTGACCCAT
TCCCTTGTTTTATTCAACTTCGAAATCTCTCTGGCTATCTTTGCAT
GCTTATCTAACTTTCACGTGACCCGGATATTGTTCTCTGCTCAAAA
AAGAACTGCAAAGAAGGAAATAGAAAAATAGCATACAGGCTCAAAG
CTATAGTTTTCTGAAAAGGGACACCACGAATACAGTAGTATCAAAG
AGAAAAAAGACTTCTTTATACTTACAGAAGGTTCATTTTCTGATTC
TACTCTTCCAATCTAGTGTCGCAAAAAAACAAAACTTACTGGTATT
AGCTGGCAAAAAAAAAAAGAATAGGTTATAATTAAAGACATCTTTT
CTCTATAAATTTTTTCTTAGAAGAAAAAGCCAAAAAAAAAAAACCT
GCTTCATACCCGCCATCAAATCTTATAGGCCTTATAGACTGCATAT
TACCGGATTCTTTCTCTCCGTTAACCCTACTATTCTTTCACTATAC
TTTAACCTAAAATAAAAAAGGAAAAAAAAAATTAAAAACTTCCAAA
AACTTAGAAATTTCAAATACAAAACAAAAAGACACGTAAAGTTAGT
GCAATATTTTTTTCTTACAATTTTTTGAAACTCGATG >YDL126C Chr 4 reverse
complement (SEQ ID NO: 53)
AGAACATTATGAAGTAAAAGGACAATCAGCACGCCTTCCAGACTTT
TAAGAAACATTGATGGAGCCATTGATATCGGCACCGTACCTAACAA
CAACAAAAATGTCTGCTCCTGCTACGCTTGATGCTGCCTGTATTTT
TTGCAAGATTATTAAAAGTATGTCACATTACTAATAAAGAGCTTAC
ACTCACACCAATGATGGCGATAGTCTCTATGTAGTACATATACATA
AAGCAGAATACTAACAATCGATCCGCTATGCAACAGGCGAAATTCC
ATCCTTCAAATTGATTGAAACAAAGTACTCGTATGCTTTCTTGGAC
ATCCAACCTACTGCTGAAGGTCATGCTTTAATCATTCCTAAGTACC
ATGGTGCGAAGTTGCATGACATCCCGGACGAATTCCTTACCGATGC
TATGCCGATTGCCAAGAGACTGGCCAAGGCAATGAAGTTGGACACT
TATAATGTGTTGCAGAATAATGGTAAAATTGCGCATCAAGAAGTCG
ACCACGTCCACTTCCATTTGATTCCTAAGAGAGATGAGAAAAGTGG
TTTGATTGTAGGGTGGCCAGCCCAAGAAACGGACTTCGATAAGTTG
GGCAAGCTACACAAGGAATTGCTTGCCAAACTAGAAGGCTCCGATT
AGAGTAATGCATGCTATGATTTCTTTTAGCGTTTTCTTTTCATATT
TAAGTTTATACATATGGGTGTGTTTGCTTCCATTCATATCCACTCT
ATAATGCTAACAATCATGTTACTAGCCATCGTTGCTATCGATACCT
TATGAACAGCTCTAACTCCTTCCTTTTTAACCTACCGCCTTTTGCG
CACTACCCGGCAACCCCTCGAATGTGAATTCGGTGGCAAAAAAGGA
CATATATCAGGGCCGTGTATTTAAGAGATCCGCAGTTGATGTAGTC
AATTAATCTAGTTCTATTAAGATTTGACTCAACGCCATTGACTCGA
TAACTGGCTCAACTACAAGACAGATATACAAATCATG >YBR106W Chr 2 (SEQ ID NO:
54) CGAACTATAGGGAGATTGGTCCTCAACAATATATTTCTTCCTTGTC
GACCCATTAGGTGTATGGGCGAGTTGCTGCAGGGAAGACAACGAAT
CCCTAGAAGCGCACTTGTAAAGATCTTGCTCTTGTAAGTAGCGATG
ACGGTCATGAAGATGAAAGGGTGCGGTTGAAGTCAAGTTGAACGTG
ATCGGATACTGATTTCTAGAAACAGGGGGGGCTGGTGTTGGTTCTG
GCGAAGCCGCTTGCTCCGACTGCTTTGATGTCACAGCTTTGGGTTT
CTCCGCTACACTGTCTACCTTAGGATTATTGATCATGACTCAAGCT
TGCGATATGTGTTGGTGTCATGCACAAGACGCCACAAATGATAGAA
CAGAAAAGAAAGTGAACTAATCTTCCAAGACGAAGAAAACCAAAAT
CCGGGATGAGTTGAAAGTCAAAAAGACTGTATATATAAATTTCAAC
TTTTGTAGAAGATGCAGAAAAAGAAAATGATATGGTATGCAGAAAA
AGAAATAAACCGCTATTATCCTCGCGGTTTGTCATTATAACAGGCA
ATTACACTAGAGAAAGCCGCACACCTCCCTCCGTTTCTTTTGCCCT
GCGAGTTTTTCCGGAAAAGAAAAAAAAAACGAAAATTAAAACCCGC
CTGCCCCGCGGGGTGGGGGAGGGGTCACCGGAACAAATCGGATAAT
CCCTCGCCTGCCTTAGATATCGTTCTGAACGGCTGAAATTATGAAA
GAAGAAGAACATCACTTTACACGGATCGCACGCCCATAATTCTTTT
TTTTTTTTTTTTCATATCTTCGACGTTTGCCACTGCCTTCTCTTTT
TCTTTCTTTTTTGGCGGCCGGTGGCCAAACGCGCCAAAACCGAAAC
GCTTATAAAATGTAGTTGCCTTGCTCTTTCATTCGATATACATATA
AGAACGCTCACTGTTATCTTACATTAGAAGCTAGAAACTATAACAG
TATAACACAGCACAAGAGAACCGAGCAGCCCGCCATG >YLL043W Chr 12 (SEQ ID
NO: 55) AGTAAAAAGAATAGTAATTATTACAATGGAGAAGATAAGATAAAGT
TAGCGGCAACAAATAAAGAACTCATCAGAGTTCTTGGCTTCTCTAT
GAGTGAAAGTGTTTCAAAGGTGTTTTTTTATGAGACAACAGTAATT
CCCCTCCTTCAGAACAGAGTGTAGTGATTGGCAGTGAAGCTATCCC
TCTTTGCCACCCAATTTTGACGGCAGTTCTCATAGCATCTCAAAGC
AATAGCAGTGCAAAAGTACATAACCGTAGGAAGGTACGCGGTAGGT
ATTTGAGTTCGTTGGTGGTTATCCTCCGCAAGGCGCTTCGGCGGTT
ATTTGTTGATAGTCGAAGAACACCAAAAAAAATGCTGTTATTGCTT
TCTCCGTAAACAATAAAACCCGGTAGCGGGATAACGCGGCTGATGC
TTTTATTTAGGAAGGAATACTTACATTATCATGAGAACATTGTCAA
GGGCATTCTGATACGGGCCTTCCATCGCAAGAAAAAGGCAGCAACG
GACTGAGGGACGGAGAGAGTTACGGCATAAGAAGTAGTAGGAGAGC
AGAGTGTCATAAAGTTATATTATTCTCGTCCTAAAGTCAATTAGTT
CTGTTGCGCTTGACAATATATGTCGTGTAATACCGTCCCTTAGCAG
AAGAAAGAAAGACGGATCCATATATGTTAAAATGCTTCAGAGATGT
TTCTTTAATGTGCCGTCCAACAAAGGTATCTTCTGTAGCTTCCTCT
ATTTTCGATCAGATCTCATAGTGAGAAGGCGCAATTCAGTAGTTAA
AAGCGGGGAACAGTGTGAATCCGGAGACGGCAAGATTGCCCGGCCC
TTTTTGCGGAAAAGATAAAACAAGATATATTGCACTTTTTCCACCA
AGAAAAACAGGAAGTGGATTAAAAAATCAACAAAGTATAACGCCTA
TTGTCCCAATAAGCGTCGGTTGTTCTTCTTTATTATTTTACCAAGT
ACGCTCGAGGGTACATTCTAATGCATTAAAAGACATG >YPR108W Chr 16 (SEQ ID
NO: 56) ATAATTGCGTTTAAACGCCTTTCCTTTTCTTCATCCATTTTCCTGG
TATTTATCTCATCGTCTCTCTTAATTCTTAATTTACTACCTTCATC
TGGGATTATGAACTGTGGATGTTCCATATCACATTCATCCTTCCCT
AAAGGACAAAATCCGGTCATGTATCTTTGACAGAAAACCTTCTTAA
TATGCCGTCTAGGACAAGAACTCCCCAGAGGACAGAATCCCATTTC
GTAATTTTCACATTTTGGTATCTTGCTAGCGGGATCTATGTGTAGA
TATTGACAATCTGGACTTTGTGTACAGTACCCGTTTTTGCTGAAGA
AGACACATTCAGGCATTTTTCGAAGATTGTATTCATGTAAGTATTC
ACATTGGTCATTCTTTTTGCACAACCCTCGAAGCCAATGTCTACAA
ACAATTTTATTCTGAAATATTGGTAACACATGCTTTTTTGGACATA
ACGGTCCCCTCGGACATGATTTAGGGCCTTCTCTAGAATTGTAAAA
TTCACAAATAGGTCTGTCAGGATCGAGTGAAAACGAATACTCTTGC
CTGAGGAAAGGTTCAAATTTAAAAGGATATTTTGCTGTATCGGGGT
GAATTAGGCTCATTACATAAACTTATTGATTTTATTCGTCCTAGAA
TACTTGTTTAAGATACAGTGCAGAAATCTTTAGCTTATAATTCAGA
AATGTGATATTTTGAAGCGATTGGAACAACAAAGTACAGTGCATTC
TATATAATTATTAAAAGTTATAGTGGACTATTGTAATAACACTTGT
TTTCCCCATATCATAACCGACGGCGCGTAAATGCATGCTCAATCAA
GTTTCCGAATAAGTTAAGTTTGCCACCCATTGAGGATTAATATACA
CAAGGCAGCATTGAAAAACCTAATTGTTAACACTACACAAATCTCT
ATTGTTGTAGTAAACTGCGCTTTTTACCACTGTTACAATAGTTACT
AATACACGCAAAAATTCTGGATAAACACGGCAAAATG >YIL041W Chr 9 (SEQ ID NO:
57) GCACGTGAATGTGATGTCTTGTGATGGCGGGTGAATTAGGACCGGG
GTCCGCTGAGTGTTGAACTTCACAAAGCATATATCATTGACATAAT
CAGAAACGTGCTTGATCAATTGTGCTATGGGGAGGTCCCGGTGCAA
GATTCCAATCATCCTTTTGTTCGTGTCACCTTTGTTCTGTGCCATC
AACGACAAATAGTGCGTCACAAGCAGCTTCATCGTGATTCTCTCCT
TCAGGTGGAAGTTCAAGAACTGCGAAATTTGAAACTTTGGGTAACA
CGATTGGATTTCCTGAAGACCTTTTGCCAATACCACAATGGCGTCC
TCGTGGTCGTCCAGCAGTTCTGTGAACTTGGCCTGTATCTTGGGAG
GATTGTGCAACTCGTACGGGTATGCGATTGATAGCAACGTTTGCAA
GCTTTTCAAATACAAAGAGTTGGTTCTCTCAATATGAGGGTTGATT
ACCGCATTATACGGTAGCCTCTGAATGGCATTAAGCCGTTTACACG
TCAATGAAAGCAACAGGTTGATTGTCTTGATCGTCAGCATGTACTC
CTCCTTCTTGGTCAGCGGCGGCCTGTATTGCAGGAAATACTCGTAG
TTTAGGGGCGCAATTGGTTTACTAGCGTAGTCCTGTATCAGCAGCT
CAATGTTCGACCTAATCTTGTAATGCTGGTCGAACGACAGTTGCGA
CAGCAGCTCGTGTGAGGGTCGCTGGCGATGTGCCCAGCGCATTCCC
CCACATTTCCATGAACGCATAATCTTCCACATACTACCGTGAAGGC
GTCTTGATACCACGTCTTCCAGTTATGGTCTTGACTCACCAATGTT
CTCAGTTTAATGTTGTTTGCTTCATACTGGCAGACATTTTCCCTTA
TTACCGTGAGAAGCGAGCGGTGGATTAATCGGGATGTCAAAAACAA
GAAAATTGCAACCTTCTTCACCTATAACCTATATGATTTGTAGGCA
GAGGAGTGGAATAAGCAACAAATCATAGTCATCAATG >YJL166W Chr 10 (SEQ ID
NO: 58) AAATCTCACTTCAGAAACGAAAAATACTACATAGATATCACCGAAT
TGTTCCATGAGGTCACCTTCCAAACCGAATTGGGCCAATTGATGGA
CTTAATCACTGCACCTGAAGACAAAGTCGACTTGAGTAAGTTCTCC
CTAAAGAAGCACTCCTTCATAGTTACTTTCAAGACTGCTTACTATT
CTTTCTACTTGCCTGTCGCATTGGCCATGTACGTTGCCGGTATCAC
GGATGAAAAGGATTTGAAACAAGCCAGAGATGTCTTGATTCCATTG
GGTGAATACTTCCAAATTCAAGATGACTACTTAGACTGCTTCGGTA
CCCCAGAACAGATCGGTAAGATCGGTACAGATATCCAAGATAACAA
ATGTTCTTGGGTAATCAACAAGGCATTGGAACTTGCTTCCGCAGAA
CAAAGAAAGACTTTAGACGAAAATTACGGTAAGAAGGACTCAGTCG
CAGAAGCCAAATGCAAAAAGATTTTCAATGACTTGAAAATTGAACA
GCTATACCACGAATATGAAGAGTCTATTGCCAAGGATTTGAAGGCC
AAAATTTCTCAGGTCGATGAGTCTCGTGGCTTCAAAGCTGATGTCT
TAACTGCGTTCTTGAACAAAGTTTACAAGAGAAGCAAATAGAACTA
ACGCTAATCGATAAAACATTAGATTTCAAACTAGATAAGGACCATG
TATAAGAACTATATACTTCCAATATAATATAGTATAAGCTTTAAGA
TAGTATCTCTCGATCTACCGTTCCACGTGACTAGTCCAAGGATTTT
TTTTAAGCCAATGAAAATGAAGAAATGCGTGATCGGAAATTACGGG
TAGTACGAGAAGGAAACTTGAGCCACCCCCCAAATTTTATTCATAT
AATAATAGGAAAAGCAACGACCTCATCTCTCGAACATTGTTTACTT
GAGCAAGTCCGATTAAGAGTAAGTTGTCGTACGTTAAATACAAATA
ATCAACAAAACACTACACAAAAACTTCTACGATAATG >YDR165W Chr 4 (SEQ ID NO:
59) AAAAAACTGAAATATAGGGTTTGTTAATCCTGGTAAAAATCTGATG
TGACATCTTCTGTACTTAGGTGGTCTTACCATGAAGTCAGCATCGA
TACAATTGATGTTATATTTCGTTGGTTCCAATATGTAAATTGCCTC
GATGGAGGATTGTCCTTTTCTTGTTGGGGAATCAATCAAATCAACA
GATGTGACATTATTTAAAAGTTCTTGGGGTGTGAGGAAAAGATAAC
TCAATATTGTCTCCACAGTTTTATCGATAATAAGAAACTTCAAGTT
ATTTTTCGTCTCAATCTGATTCAAAACCCCTATCAAATAGTTCCTC
TGTAATTCAATTAAATCAGACATCGTTCCGCCAAACCAAGCATGTG
CACACTCTTCTACTTTGTCAGTGGATTCTAATTATTTTCAAGGTCT
TACAAAGCAATGTCTTACTAGTTTCAATAGGGACTTAAACTGATCG
TTTAAGCAAAAGAAAAACACCGTGCTGTGTGACCATCTACCAATAA
TTGCGAGAACATAAGACGACGCCGCCATTCACCTCTCTATCACTTA
CACCATGTTATAGGGATAAGTGTGTAGTCGATCGGCTATTCGATGA
TTTTTAAAGAGTGCCTGTAATAATAGCTGTATCGCAAGTAAGGAAT
ACAAGATTACTGATATTTTGATCCAGGAGGATGGACATGTTAGATA
GAAAAACTAAGAGGGCTACTACATCCGGTTTAGTATTCTGTTATTG
GGAATGCTTTTCCCAAGACGCAAGCAAGAGGTGACACATAAACATA
GTTTTCTTATTGTATCGTTACATATTCTTATTTTACGCTGCTGTCA
TTAAATATGGCACTTATCTCAGATACATTTCTTTTAATCAGCGAAA
AAAATGGCGAGGTGAAAAAAATCGATGAGATGAAAAAATTTTTTAC
TAGTAAAGGCGTATTGATGATATATTAGCAGCTAAAGTATAGCAAG
GTTTGTCAATTTGAAGCAGTCCGCGTGTTTTACAATG >YLR330W Chr 12 (SEQ ID
NO: 60) CTAACTATAATAACTGGTAGCTTTGTCACTCGTACCAGGAAAAGTG
AAGATTAAACTGAATTTTAAAATGAACGATCGATTAGTTACGGAAG
AGCAAGAGTTGTTTACAAAATTGCGCGAGATTGTAGGTTCAAGTAT
TCGCTTTTGGGAGGAACAACTGTTTTATCAAGTTCAAGATGTAAGC
ACCATAGAAAACCACGTCATTCTCAGTTTAAAATGTACAATTTTAA
CGGATGCTCAGATAAGTACGTTCATAAGCAAACCCAGAGAGCTTCA
TACGCATGCCAAAGGATATCCTGAAATCTATTACCTTTCCGAGTTA
TCAACAACTGTCAATTTTTTTTCTAAAGAGGGAAACTATGTCGAAA
TAAGCCAGGTTATTCCTCATTTTAATGAATATTTTTCCTCTTTAAT
AGTGTCTCAATTGGAATTTGAATACCCGATGGTCTTCTCCATGATT
TCAAGGCTCCGATTGAAGTGGCAACAAAGTTCGCTCGCTCCGATAT
CCTACGCCCTAACGAGCAATTCAGTACTTCTTCCAATAATGCTTAA
CATGATTGCCCAAGACAAATCTTCAACAACCGCGTATCAAATTCTG
TGTCGAAGAAGAGGTCCTCCAATTCAGAATTTTCAAATTTTTTCCT
TACCGGCTGTAACGTACAATAAGTAGCATGCATAAAATATAATTTA
ATCAAATACTTTTGGGCAATTAAAATTTTAGTTAACAATAGTTATG
CAATGCGCTTTATGTTCATATGATACCGTTTATAAGCTATTGCCAT
ATCCTTATCTTATTGCTTCCAGTAGCCTCGAGTCGACCACTAAAAA
GATGTCACTTAAGACGGAAATTATGTAGCTGCACTTCTTTTTTAAC
AAGTTCGGTCGGCCCTTCAAGTTCTCCTTTCTAAAGCCTCATTATT
TATTGCGTAGATGCTAAATGTTATCGCGGTTTAGCTTGCATGTTAC
GTTTCCGTTTTAGAACCTGGTCGAGTAGCGAATAATG >YDL058W Chr 4 (SEQ ID NO:
61) TTCGCCAACCATCATACCCGAATGTTTCATTAGCGAACTGAATGAG
AGCATGCCCTGGTATCAGCTTAGATAAGTTGTGCTTCCCATATTTA
TTATTGTGGTAAATATTGTACGTGTACCTTTCGATCTTGGACTGTA
GAAGCCCAATCCTCTGCACCGACCAAGCGCTGGCAGGTCTTCCATT
CCAATCTTCGATGATTTGGAACTCTTTTATATCCAAGCCCGGTGCT
GTGCCGTATGTAGTCGAATCATACGATATGCTCGAACTGGGCTTCG
CTTGTATCGTCATTATTTTGTCTGTTTATCCTGAAATATGCTCTTC
TCCTCTACGTAACCCTTTAATGGTCCAAAGCATGTGCTAAACTGCC
ACAGACTCCCTCAATGATGTTCTTATCCTTCCTTGCCGTTACTGCT
TCCTATCATATCTGCCCTTGCCAAACTGTGGCTTCCTCTAATCAAT
TTGCCCCAGTACATTGAAAAGCACGTGACGTGACACCAACTTGAAA
GACATTAGTGTCCAACGCGATATAATGGTCAATAAAATACCCAATT
TTATCAAGTGAACATGATATTATTCAATACTATAGTACTTTAAAGA
AAACTTCAACAAGTTGCTTCATTGCTATATTATACAATTATGGGTG
ATGCCGGTCGGCTAACCTTTCTTTCGCAGCGCCACATACAAGAACA
TATGGTAATTACTAAGAACTTTTTGGAAACTATTTCCATGAAGAAA
ACAAAATTTACTGGCTTCTGTCTTCGAAGTAAATTTAGCGAACTTG
ATACTTTCGTTTGGACAGTCGTTTAGATTTGCAACTGTCTGTTCTT
ATTTCTTGCTATTATTACCAGAAATAAAGCCTAAAGCGCGTGGTTC
TTACGAAAAAAATAGACCTTGCACATTAACAGGGTTTAGACATCAC
AGAGACTGAATACTGGAGAGAAGACAATTCTTGTTCACGACTTCGA
GTGATTAATATCTATAACAACCCTCCTATCAAGAATG
>YHR012W Chr 8 (SEQ ID NO: 62)
TGAAAGGCAAATTTACGCTCTTCAAGACACCAATTTAGGATTGGTG
GCTACCGCGGAGATACCTTTGGCAGGTCTAGGGGCAAACAAAGTTC
TAGAGTTAAACTCGGGAGAATGTTCTAAGAAACTAGTTGGAGTAAG
TAGATGTTACAGAGCTGAGGCAGGTGCCAGGGGAAAAGATACGAAA
GGTCTCTATCGCGTTCATGAGTTTACTAAAGTAGAATTATTTTGCT
GGAGCAAGCCAGAAACCAGTGCAAAGGTCCTTGAGGAAATAAAGCA
ATTTCAGATTTCTGTAGTTGAGGAATTAGGAATACCGGCCAAAGTA
CTAAATATGCCATCAAACGATCTTGGTAATCCCGCTTTCAAGAAAT
ACGACATCGAAGCTTGGATGCCAGGAAGGGGGAAATTTGGCGAGAT
AAGTAGTGCCTCCAATTGTACCGATTTCCAAAGTAGAAGATTAAAT
ACAAAGTACAGGGACGATAACACAGGAAAGCTAGAATATGTGCACA
CGCTGAATGGCACAGCAATGGCTATCCCAAGAGTGATAGTAGCACT
AGTAGAAAATTTCTATGACCCAAGCACCGGTAAGATATCTGTTCCT
GAATGTTTGAGGGAGTTTATGAATGGCCAACGATACATTTAATAAG
TAAGAAAATCTTGTAAATATGTAATCTTATGTTAACCGTAGCATCA
TTGTGTTGGACGAAATCCTCTGTGATGTTCTGTGTGATTGAAATTG
TATGGTATTTTCGATCATACGAGTATTTTTCCTAAAGATGAAAGAG
AACTCGATTCCTAAACAGTTGTTTGTAGGGCATAACGTTGTATCGT
AAATACTTACCATACGCTTAGGTATATTCCTTTAGATTTCCGTAAC
ACCTTTCAGTTTTACTATTCCGCGTGGCGAGGTAAAGATCATAAAA
ATACAAGTTTGTTATAGCATTTCCTGCAAATAATTGCTGTAACTAG
TGGCGAAAAGGTCATAGAATTATTCGCCTAAATTATG >YKL053W Chr 11 (SEQ ID
NO: 63) CCGGAAGTAATTTTATCAATTATAGTTTCCAAATCATCATATTCTT
GAACTATATCAATTAAATCATCACTCGTCCAGTCAGGGAACAATTC
TGTAAGCGTATCTATTTTGGACTTTAGCGCAGGATTTAGTTTTTTA
CTACTATGACTATTATGATTAGACTTCCTAAATTGTGTAGACATTA
TATATATATGATGATGTAGATTATATTGAGAAAACGACCAAACAAA
AATAAATATGTAAGTACTTTATGATGGTGATTTTTTTGGGGTAATT
GGTAAGAGTTTTTCTTGTAGCTTCAAAAGATCAAATATAAAATACG
AATGTTTCTGAAATTACTGTAAAAGACTTAGGAAAAGGAATGAAAC
GTTTATATGGGAGAGAACGAAAATTTTTCCTTTTCCCGCCACTACT
TTTTGCCACCATTTCCCGTTATTTATTTGGGCGAGTGCTCACCTCC
GACGGAATCACGGGTGACGGATATCCTAAAAAGGTAAAGAATTGAA
ACAGATCTATTAATATACGGATCACTATACATTATTATAAGTCGAT
ATTGCAGTATTAGTATTTACATGTTGAATAATGCACATTCTGTGCT
AAAATATATACTTCATTTGTCAACTTCTTTTAGTTTGCCGCCATTT
TCAAACGGCGCCTCCTCCCTAGCTTCGTCTAGAGCAGGCTTAATAC
CTTGTTTGACGAGAGCTGCGTTGACACATGTAGTATAAGCGTACCA
TTGTTTTGAGCACTCGTTCTCAACGGATTTTCCCTTCAGGAATTTT
TCGCTATACCATTCATTAAAACAACTATCGTATTTCGTCTTCAGGT
CAGTGCATTCAGGCGCAAAACTAGCTGACATTATATTCCCCATATA
AAACTGGTATTATTGTTATTGTAATTCAAGTTAAAACACAAACAGA
TGTTTTCTAATGACCTCTCCCGGCATATATCTGGCGCACTTAGAAT
AAATTTCTACATGCAAATACTCTGAATATTCTGAATG >YKL196C Chr 11 reverse
complement (SEQ ID NO: 64)
TTCTTCTTCTCCATTATTATCTTCAGTAGTTGTCTTGTCATCAGAT
TGTCCCTGTTGCGATTTTTCATCATCTGAATGTTCTGGATCTTTTT
TTTCAGAAGATCCAGCATTGTTGTCTAGTGTTTTGTCACTAGTATT
TTCTTCCGAGTTCTCAGAACTTATGTTTTCATCCTTTTGATTGTTA
GCGGTGGCATTGTCTTCATTGGATTGCTCATCATCTTTGGTTGATA
CCTTTGATTCATCAGTATCTTTATCTTCGGAAGACTTGTTGTCTTC
TTCTGCTAGAACAACCAATTCACCATCTTCAGCTACTTTTGAAGCA
CCAATTTTATCACCCCCTAATTGAGGGCCAGCTTCAGTTTCATCGT
TTTTGCTGTCGTTGTCATCTCCATTTTCATCGCTTTTGCTGTTGTT
GTCATCTTCATTTTCTGGTGATTGCTCACCACCACTGGAAAGCTCT
TCCTCTGCGGTTTTATCGGATTCTACCTTCCTTGAGGCGAAAAGAG
GCTTCCTATTAGGAGCAATAAAATATAAAGCACCAGCCATAGAAAG
AATCCCCATTATAAAGCCCGCTGTTTTTTCCTGATTGGAGTTCCTA
CCGAACTGAGGGGAGGACGCCATGAGACGTCTTGTTTGGTGTCGGC
ATAACCCCCTTGCCACTTGAATTGACGGCCTGTTTCTGCACGCATT
CCTGACGACTAAGTTGCGAAGCATTTTACTGATAATATACACTCTT
TGGATCGAGCCTACTTCCAGTTGGTAATTGGTGTTCCACAATTTCA
GCATTATATGTTTTTAAACCAAAATTCGGCTCCTTTTCCCTTTTTT
TCTTATTGGGTGGCGTGCCGTACAGAACGATTGGCTTGGTGTGAAA
TCAAGAGCAAGCACAATAGATATCAACATGAACAATATACAAAAGT
CTCTGGCACAGTTTGACTGCGTTAGACCAGGCTAGGGCATTTCTGA
AGCTTTACGTATCACTAGAGAAGTTATTTTGGCAATG YDL103C Chr 4 reverse
complement (SEQ ID NO: 65)
GAAAGATGACCTCTTTCAAACGCAGTACGCAGTTTGTTGACCATAT
GCGGATAGGTGACGTATATTTTCCAGAATGGTAATTTGGTATCTCC
GGAATAACCCCAGATAGACTGTTTAGATACAACTTCAATCGAATCA
ATAGCGTGGTCAAATGTTTCGTTTAAATAGTGCACAAACTTGTTGA
TTTGCTCCTGATCGTTAGCGTCGGAAGAATTGGGCGCTGGGACGTA
AAGATAGTTCTTGAACCCTGTAACATTACAAAGTACAGAGTGTCCT
TCACTAGTGACACCAAAAAACCTTACCACGGTAGATGTATTTTCAT
CTTTGATACCATTCAGTACGCTCTGTTCCGCATCAATTTGTTGGAA
AGAAATATCATACAAACTTGGGTCAAAATCGGTTGGAAGTTTCTTA
CGCTCGAATGATGACAGGTCATGCTCTTCTTGGTCGGCCATATCAT
GTTCCATTTGCGATACGTCTTGTTCAAAAGTAGACTCTAATTGCGT
ACCCATTAAATCTGTATCCTTTGCTTTGAAGCCTTGACTATTATAT
TTTCGAAAAGAATCACTCGGAATAATCTCTATTGTTGAAACAGGTT
CACTTCCAACACCATGATCTATTGATTGCCGTTTGATTTTCTTTTC
CAACTGGGGAGTATCCTCGTCATCGATCTTCACATCAACCATGGGA
AGGGATCTTTTTTCACTCATTGCTGTGCGTATATGTATAAAGATTA
ATGCTTAATAGCAAGTGCTCAATATCTATTTTCCTTTATCCTTGAA
AATTGCCTTAGATCAGTAGTACTGAGTGTTTGTTCTTCATAAGCGA
ATAGCAGATGTCCTCATTGAACATTTTATAGAATAAAAAGTTACGC
GTAATACGAAAAATTGCCTATTACGTTAAGGCATACTTTTACCACA
CAAAACCAAGATCAACCGTCAAGCAAGAACGCTGCAAAGAAACGCA
GATATAAAGGAGAACACCAGATCGAACTGCAACTATG >YLR427W Chr 12 (SEQ ID
NO: 66) TCTTGAATGTCGACATATGCCCATCATCAGTGAGGAATACTCGCGT
GAAGGATCTCATCTGCGACCTGAGTGATGATGAAGAAGTTGCAGCA
CTACTGAATTTGTTGAAAAGAAAGTATAAAAATGAAATCCGATTGA
TTGTGAACAATGCTGGAGTAAGGGCCAACTTCACCGGATTCAACGG
CATGGAACGGGATAATCTTGATAAGATTTTTAAGATAAACACATTT
GCACCACTTCAGTTCATTCAAGAACTGGCCCCTAGTAGACACTCAA
CTAGACAGTGTTATATTGTCAATATCGCAAGCATTTTGGGCATATT
GACGCCGGCCAAAGTAGCAGCTTATGCGGCGAGCAAAGCAGCATTG
ATAGCCTTTCATCAGTCGTACAGTTTTGAATTGCAAAACGAAGGCG
TAAGAAATATCAGAACACTATTAGTGACCCCAGGGCAACTGAATAC
GGAAATGTTTGCAGGATTCAAGCCTCCTCGCCAGTTCTTTGCCCCT
GTAATAGACATTACTACTCTAGCTGCCAAGATAGTCCGTTATTGTG
AGCTGGGTCAGAGGGGACAGCTAAATGAACCCTTTTATTGTAGTTT
TGCTCATCTTTTGATGTGCGTACCTTATTCGCTACAACGCATTGTA
AGAAGCTTCTCTCGCATAGATTGCTGCCTCCCGGACGAGTAGAAAG
CTATACATAGTAAATAGAATATTTATAGATGAATCCGAGTAGACAG
CCTTGGTGTTTGCTTATACATTCTTTGCAATTATGCACGCCCTCAT
TACCGTTGCTCATATTTTTGGGCATTAATTGTATTTTGAAAAGTGC
TCGTTCAGGCCCGTGTAAAAAAAGGTGATGAAGCAAACATTATAAT
AAACACTTCTGAGAAGCCACGTGTAGAGGGGTAAGCTATATCGTAA
ACACCGTTGGATGTGGACCACGGTGCACTGATAAAATAAAGATATA
GTAGTAGAATCTGTTACTAATCTTAACACTTTTGATG >YLL020C Chr 12 reverse
complement (SEQ ID NO: 67)
GCTGTAGATAGATATAGAGAAGCCGCCAAAACAGAACTAAGAATTC
TACAGACTATCCTGAATAATGACCCTCAAGGTCAGTTCCAGTGCCT
CTTGCTAAGGGAGTGCTTCGATTACAAAAATCACATTTGTTTGGTG
ACAGATCTATACGGCAGGTCCATTTACGATTTTATGTGCTCCAACG
GCATTGCCAGGTTCCCCGGCTCTCATATTCAGGCCATTGCCAGACA
GCTAATCAGATCTGTCTGCTTCTTGCACGATTTGGGCATAATACAC
ACGGATTTGAAACCAGAAAATATCCTGATTTGTGACGAAACCCATA
TTGCTCAAAAATTGCCTTTGAAAACCGTACAGTCGCTAAGCAAGAG
ACGCCGTGAAGCCAGTAAGGGGAAGCGCAAAATCTTGAAAAATCCA
GAAATCAAAATCATTGATTTCGGTAGCGCAATTTTCCATTACGAAT
ATCATCCTCCTGTAATATCCACTCGTCACTATAGAGCCCCGGAAAT
TGTCCTTGGCTTGGGCTGGTCGTTCCCCTGCGACATTTGGTCCATC
GCGTGTGTCCTAGTAGAACTGGTTATTGGCGAATCTCTTTACCCCA
TTCATGAAAATCTAGAACATATGGCTATGATGCAACGAATCAATGG
AACCCCTTTCCCCACAGACATTATTGATAAGATGTTTTACAAATCT
AAACATAAATTGGGCAACTCTCCATCAGACCTAAATTCAACGGTGA
TAAAGCATTTCGACAGAAAAACTTTAAGCTTACAATGGCCTGAAAA
AAATAAACGCGGAGATACCATAACCACCGAAAAGTCAATGAAAAGG
GTCTTGCAGTCATGTGATCGCTTAGATATTTACATCTCAAAGGTCT
TGAAACAGGATTACGGCGATAGCTTAAGCATCAATTGGAATCTACC
GCCGGAGAAAAACTGGTCTTTGATAAATTCGAAATTGGCATGGAAA
AGACAAACACATTCCTCTTCTTCCTCAACTACTGATG
Example 2
Ratiometric Screening Assay (Luciferase and
.beta.-Galactosidase)
[0105] In this example, the ability of a substance to modulate the
chromatin modeling function of the YCR052W gene is determined using
a ratiometric assay.
[0106] S. cerevisiae strain W3031a cells are transformed with a
plasmid comprising a hybrid which includes the responsive promoter
YOR387C (SEQ ID NO: 1) operably linked to the E. coli lacZ gene
along with a plasmid comprising a hybrid which includes the
baseline promoter of YHR082C (SEQ ID NOs: 52) operably linked to
the Photinus pyralis luciferase gene lacking the peroxisomal
targeting sequence (see Leskinen et al., Yeast 20:1109-1113
(2003)). The cells are grown, in microtiter dish wells containing
100 .mu.l culture, while shaking at 30.degree. C., to exponential
phase and the culture is split in two. One culture is contacted
with a substance to be tested for the ability to modulate chromatin
modeling and the other culture is not. The cells in each culture
are further incubated for another 2 hours.
[0107] 100 .mu.l of 1 mM D-luciferin in 0.1M M Na-citrate (pH 5.0)
are pipetted into each well. The plate is shaken briefly and then
the luminescence is measured wuing a Victor mulitlabel counter in
luminescence mode (Perkin-Elmer Wallac; Turku, Finland) using a one
second counting time. After the measurement of the luminescence,
CPRG (chlorophenol red .beta.-D-galactopyranoside)
.beta.-galactosidase test buffer (for example, 50 or 150 mg/ml of
CPRG in HBSS (Hank's balanced salt solution)) is added. Then, the
expression of the lacZ reporter gene is measured by measuring the
chlorophenol red reaction product, optical absorption at 540 to 580
nm, after another 2 minutes to 24 hours of incubation.
[0108] The levels of expression, in each culture tested, of the
lacZ gene is normalized to the level of expression of the
luciferase gene. The normalized level of lacZ expression in the
presence of the substance being tested is then compared to the
normalized level of expression of the lacZ gene in the absence of
the substance. If the ratio is above or below 1, this indicates
that the test substance is modulating the chromatin modeling
activity of the cell.
Example 3
IC90/MIC Calculation
[0109] In this example, a baseline promoter of the invention (SEQ
ID NO: 53) was used to determine the IC90 of various known
anti-fungal compounds. The IC90 value determined was then compared
to a visually determined MIC value.
Setting up the Plates:
[0110] 1. Picked a single colony of yeast Y229-1 (P.sub.CDC48 (SEQ
ID NO: 53)-PHO5-Renilla Luciferase) and inoculated 10 mL YPD (1%
yeast extract, 2% peptone, 2% dextrose) containing 100 .mu.g/mL
G418 (Geneticin, GIBCO, Invitrogen Corp.). [0111] 2. Incubated the
cells overnight at 30.degree. C. with shaking (250 rpm). [0112] 3.
Measured optical density of the culture at 600 nm (OD.sub.600) and
diluted an aliquot to OD.sub.600=0.1 in YPD. Made an additional
1:100 dilution of the 0.1 OD.sub.600 culture in YPD. [0113] 4.
Transferred 12.5 .mu.L of serially diluted compound or no drug
control in 2% DMSO to one well of a black/clear bottom sterile 384
well plate. Covered with a sterile lid. [0114] 5. Added a 12.5
.mu.L aliquot of diluted Y229-1 cells to each well of the 384 well
plate. Final DMSO concentration was 1%. [0115] 6. Covered the 384
well plate with a sterile lid, placed in a plastic bag to limit
evaporation, and incubated for 24 hours at 30.degree. C.
Preparing the Luciferase Assay:
[0116] A Promega Renilla Luciferase Kit was used in these steps
(E2820, Promega Corp., Madison, Wis.). [0117] 1. Add 12.5 .mu.L
1.times. Lysis buffer per well. [0118] 2. Add 25 .mu.L Assay
Reagent per well of 384 well plate. Read light output immediately
on an appropriate device (e.g., LeadSeeker, Amersham Biosciences,
Piscataway, N.J.).
TABLE-US-00006 [0118] TABLE 4 Correlation of IC90 calculated from
luciferase light output and MIC determined visually 5FC AMB CSP FLU
TRB VOR IC90 1.9 0.3 0.02 0.3 1.4 0.008 (.mu.g/mL) MIC 2.5 0.3
0.015 0.2 1.56 0.015 (.mu.g/mL) IC90, inhibitor concentration at
which there is a 90% drop in signal relative to the uninhibited
control; MIC, minimum inhibitor concentration at which there is an
approximately 90% drop in cell number relative to the uninhibited
control; 5FC, 5-fluorocytosine; AMB, amphotericin B; CSP,
caspofungin; FLU, fluconazole; TRB, terbinafine; VOR,
voriconazole.
Example 4
High Throughput Screen
[0119] In this example, a baseline promoter of the invention (SEQ
ID NO: 53) is used in a high throughput screen to discover
compounds with antifungal activity.
Compound Preparation
[0120] 1. Prepare compounds in 5% DMSO. [0121] 2. A control
inhibitor, amphotericin B is prepared at 25 .mu.g/mL (in 5% DMSO)
to produce 5 .mu.g/mL (final concentration).
Culture Inoculum Preparation
[0121] [0122] 1. Inoculate 10 mL YPDU (1% yeast extract, 2%
peptone, 2% dextrose, 80 .mu.g/mL uridine) supplemented with 0.1
mg/mL G418 (from 50 mg/mL stock) with yeast strain Y229-1
(P.sub.CDC48 (SEQ ID NO: 53)-PHOS-Renilla Luciferase). [0123] 2.
Grow Y229-1 overnight at 30.degree. C. with shaking (250 rpm).
[0124] 3. Measure the optical density of the yeast culture at 600
nm (OD.sub.600) and determine volume of yeast culture needed for
OD.sub.600=0.1 in 5 ml YPDU. [0125] 4. Further dilute this 5 mL
culture 1:20 in YPDU (this volume is sufficient for approximately
5000 assay points).
High Throughput Assay Set Up
[0125] [0126] 1. Add 5 .mu.L compound or 5% DMSO to a sterile 384
well plate (e.g., Matrix Technologies, Hudson, N.H. catalog #4331).
[0127] 2. Add 20 .mu.L of dilute inoculum to each well containing
compound, amphotericin B or DMSO. [0128] 3. Cover plate (e.g.,
Matrix Technologies catalog #4320) and incubate 24 hr at 30.degree.
C. [0129] 4. Prepare 1.times. Lysis Reagent from Promega Renilla
luciferase kit (catalog #E2820) by diluting 5.times. stock in
deionized H.sub.2O. [0130] 5. Add 12.5 .mu.l 1.times. Lysis Reagent
to each well. Incubate at room temperature for 1 hour. [0131] 6.
Prepare Assay Reagent as described by manufacturer. [0132] 7.
Dispense 25 .mu.L Assay Reagent into each well. [0133] 8. Read
immediately on an appropriate luminescence detection device.
Example 5
Ratiometric Dual Luciferase Assay
[0134] In this example, a Renilla luciferase and a firefly
luciferase reporter is linked to a gene responsive and a baseline
promoter, respectively, and used in a ratiometric assay to
determine whether a substance inhibits fungal cell growth.
[0135] Strain construction. A double recombinant strain comprising
a chromsomally integrated YOR387C promoter/Pho5-Renilla luciferase
hybrid and a chromsomally integrated YHR082C promoter/Pho5-Firefly
luciferase (--SLK) hybrid is constructed. The yeast strain, Y229-1,
is transformed with plasmid pSPRT190 (SEQ ID NO: 87), comprising
gene responsive S. cerevisiae YOR387C promoter (SEQ ID NO: 1)
operably linked to Pho5-Renilla luciferase. The YOR387C
promoter/Pho5-Renilla luciferase hybrid integrates into the Y229-1
cell's chromosomal TRP3 locus. The pSPRT190 plasmid comprises the
KanMX gene for selection. The recombinant strain is, in turn,
transformed with plasmid pSPRT50 (SEQ ID NO: 86) comprising
baseline S. cerevisiae YHR082C promoter (SEQ ID NO:52) operably
linked to Pho5-Firefly luciferase (-SLK). The YHR082C
promoter/Pho5-Firefly luciferase (-SLK) hybrid is allowed to
integrate into the Y229-1 cell's chromosomal HO locus. The pSPRT190
plasmid comprises the KanMX gene for selection
[0136] Assay. Test and control double recombinant yeast cells are
mixed in growth medium and diluted to OD.sub.600 of 0.2. Two
experiments are set up: (i) double recombinant strain contacted
with the substance being tested for the ability to inhibit fungal
cell growth and (ii) double recombinant strain contacted only with
a blank. Then, 25 .mu.l of the diluted yeast strains are added to
microtiter wells containing a compound to be tested for the ability
to inhibit fungal cell growth or a blank (10 .mu.l compound
[dissolved in 100% DMSO] per well). Yeast and compound are
incubated for 3 h at RT, followed by the addition of 25 .mu.l of
firefly luciferase reagent (Promega, Madison, Wis.) to each well,
and incubated for 1 hour. Luminescence (firefly luciferase
reporter) is determined using a Viewlux plate reader (Perkin Elmer,
Wellesley, Mass.).
[0137] After reading the firefly luciferase counts, using a Victor2
plate reader (Perkin Elmer, Wellesley, Mass.) equipped with an
injection system and interfaced in a Thermo-CRS stacker-based
robotic system (Thermo-CRS Ltd., Burlington, Ontario, Canada), 25
.mu.l Renilla luciferase reagent (Stop & Glow, Promega,
Madison, Wis.) are added to each well individually, and
luminescence is determined immediately (within seconds, due to the
short half-life of the Renilla luminescence).
[0138] The relative levels of expression from the YOR387C and
YHR082C promoters, in the presence and absence of the substance
being tested for the ability to inhibit fungal cell growth, are
compared.
[0139] In this assay, the compound being tested for the ability to
inhibit fungal cell growth is selected if the responsive promoter
(YOR387C) expression level (Renilla luciferase) decreases or
increases in relation to the expression level from the baseline
promoter (YHR082C) (fire fly luciferase) when the relative
expression levels in the presence of the substance being tested are
compared to the relative expression levels in the absence of the
substance being tested.
[0140] Alternatively, the plasmid pSPRT47 or pSPRT192 can be used
to construct the assay strain. Plasmid pSPRT47 comprises a
Pho5-Renilla luciferase construct to which a promoter can be
operably linked, an HO targeting sequence, and the KanMX gene for
selection. Plasmid pSPRT192 comprise a Pho5-Renilla luciferase
construct to which a promoter can be operably linked, a TRP3
targeting sequence, and a hisG:URA3:hisG consruct for selection and
counterselection (e.g., using 5-FOA).
[0141] The present invention is not to be limited in scope by the
specific embodiments described herein. Indeed, various
modifications of the invention in addition to those described
herein will become apparent to those skilled in the art from the
foregoing description. Such modifications are intended to fall
within the scope of the appended claims.
[0142] Patents, patent applications, publications, product
descriptions, Genbank Accession Numbers and protocols are cited
throughout this application, the disclosures of which are
incorporated herein by reference in their entireties for all
purposes.
Sequence CWU 1
1
8811006DNASaccharomyces cerevisiae 1gccagggaag cctatatctg
attccctgtt tcataatcca atgcagccac tagcttataa 60ttatttgaac tatttgtcga
acatcacagt aataaaatcc ccagaaagtt ccacttgctg 120catattggca
cctgttgatt cactctccat cacttttttg ttagccgccc agcctagaaa
180gtctttaaat acatctgaaa tttttttttt tttaacagtg cacccgtgca
tcatacctca 240tgcaaggtac ctttttttct caaaggtatt gtcttccatt
gaagtggcac tatggcatga 300tgaaccctga gcatttctga attcaacaga
accaaattgt ccagaaataa atctgtccga 360catgaattat gaaacttttt
ttcaattaag tgaagagaat tttgcagcgt cttaccatta 420ttttgaccca
ttggtcgcat gtttgcgctt tgacttcgag aaccatgtta aagcttactt
480gtacgacaac caatgaagta tattacggca gtttttttgg actgggtcaa
aaaaagtgtt 540gcataatcaa atcaggaaca cattaaaatg ttgtaaaatt
tgtcttagta tcacctgagt 600ggttattcat tacgtactac tgtcaaaata
ttcggatctt tcctaaacgg gcttttgaat 660tagtgttgct tctattcctg
gaatggaagg taatactttc atgcattctc gcttttcgga 720cttttaacaa
taaattaaaa acaatgatac tttcatcaac tacctataca ccctgcgggt
780caattatttt tttttcgaat aaccgctgag gttggaaata tagaaacata
ttccagatct 840gtattttcag ttgctagaaa aaggttaata taatcattaa
ggttttcagc atataacagg 900tataattgat atataagcat cgtaattttc
attcaaaatg gagagctact gcttctgata 960gattgtacaa tctcaagaaa
tcaagaacaa caaccatacc atgagt 100621003DNASaccharomyces cerevisiae
2ttgtgtttag acttaagtat gaaaatttta tgtatgagct gtggctatgt atccgctggc
60aaataggtct gctttttcta cactttccac cctcaaccta acagagcccg ctggcacaaa
120taatcgatag taggacaaca gagctactcc ttcttatgcc ccgccccttt
gagcttgttg 180tattgctctt gatagttgtg tttttcactt tcatcagcat
cggtagtctt gccgtccttg 240tcttgactag ccattttctt aaaagcgtca
ctcactacct ttgcatcgcc ctgcaatttt 300tcctttgagc tctgaaatat
ttcgctcact ttttgttggt ctgtattcat tctggatgtc 360ttggttgtag
aaatttcttt tattggttca ttaaagtcaa ggtaaatggc gagaactaga
420atagagtttt attcttttta ccgttatata gataattcta gccgggggcg
gtcgcccctg 480agattcccga catcagtaag acatagtact gtacgattac
tgtacgatta atctatccac 540ttcagatgtt caacaattcc ttttggcatt
acgtattaat acttcatagg atcggcaccc 600tcccttaagc ctcccctaaa
tgctttcggt acccctttaa gacaactatc tcttaacctt 660ctgtatttac
ttgcatgtta cgttgagtct cattggaggt ttgcatcata tgtttaggtt
720tttttggaaa cgtggacggc tcatagtgat tggtaaatgg gagttacgaa
taaacgtatc 780ttaaagggag cggtatgtaa aatggataga tgatcatgaa
tacagtacga ggtgtaaaga 840atgatgggac tgagagggca attatcatcc
ctcagaatca acatcacaaa catatataaa 900gctcccaatt ctgccccaaa
gttttgtccc taggcatttt taatctttgt atctgtgctc 960tttactttag
tagaaaggta tataaaaaag tatagtcaag atg 100331003DNASaccharomyces
cerevisiae 3gaggtttcaa catgccaaca atggaagtgg ttaagagacg ttttctggtt
tctgggcagc 60tgttacatag aatctctgat ttacatgctc aagtattcgc aaagatcggt
atgatgggaa 120aataggtgaa ttttgggata atgactgaga ttataccgtt
gttaatgcta taatattaat 180tatacagaat atattagaag ttctcctcga
ggatatggga acccacagaa gcgaatcgat 240gtttctacat aaaagaaaaa
aataatctca ttttatcttc ggttttgaat cttttcattt 300ttttttccta
ttacattatc aattctggca tttcagtgtt tataaaaaga tcaaattgct
360gtttgaaact cattgcgcta attccataat tttgcaccat gtatgctcat
gactccatat 420aggacaacat actggtagga atattatcaa gtgaacgata
acggttcttc tcccaacaag 480aagtaagcct ggtgagtgta ttttgaagct
ttttggaggg gcggccacgg aataagatat 540gaacacatac tggcaattgg
gctgagatct gattacattg tcgacagatt cccatgaagg 600tgtttgagag
aattttgatt gagtcgttat acaaattgct gtggacacgt acgcataaag
660tagttcgggc agctatgttt gccacgtagt agctacaata ttaagcatag
cagactaaat 720gcaattggct gtgttgaaag aaacttctga actgagattc
aagtaacgtg attgtgtgcg 780ccattttaca caggctgcgc ggcaaatcaa
aaccaaagat acttctccgg aggtatacgt 840gttgcatttt caacagacta
tacgagtttt ttctgatcgt tgtatattta agtgcagctt 900ggacttacaa
gctctatact aggataatga tctcattgga ttcaagagaa agaaactcta
960tactggcgcc aaattagcag tgtcaaattt cgaaaaggtg atg
100341003DNASaccharomyces cerevisiae 4ttcttttttt tttttttttt
ttctttttgg gaacggggga aaaggtctca ctagtatata 60aaggaaagag agtacagcac
gaatgagcca gctgcaggaa aggggctagg ttaaaaaata 120aaaacccagc
agaaagcaaa caaccaaata taatttagaa atggacagag accatattaa
180tgaccatgac catcgaatga gctattccat caacaaggac gacttgttgt
taatggtttt 240ggcggttttc attcccccag tggccgtctg gaagcgtaag
ggtatgttca acagggatac 300actattgaac ttacttctct tcctactgtt
attcttccca gcaatcattc acgcttgcta 360cgttgtatat gaaacgagta
gtgaacgttc gtacgatctt tcacgcagac atgcgactgc 420gcccgccgta
gaccgtgacc tggaagctca ccctgcagag gaatctcaag cacagcctcc
480agcatatgat gaagacgatg aggccggtgc cgatgtgccc ttgatggaca
acaaacaaca 540gctctcttcc ggccgtactt agtgatcgga acgagctctt
tatcaccgta gttctaaata 600acacatagag taaattattg cctttttctt
cgttcctttt gttcttcacg tcctttttat 660gaaatacgtg ccggtgttcc
ggggttggat gcggaatcga aagtgttgaa tgtgaaatat 720gcggaggcca
agtatgcgct tcggcggcta aatgcggcat gtgaaaagta ttgtctattt
780tatcttcatc cttctttccc agaatattga acttatttaa ttcacatgga
gcagagaaag 840cgcacctctg cgttggcggc aatgttaatt tgagacgtat
ataaattgga gctttcgtca 900cctttttttg gcttgttctg ttgtcgggtt
cctaatgtta gttttatcct tgatttattc 960tgtttcattc cctttttttt
ccagtgaaaa agaagtaaca atg 100351003DNASaccharomyces cerevisiae
5gaagttacaa ataataacaa caaagttaat aacagtagta gcaaccctga tatttccacc
60aactcggtag ttcataacgc aatgcaattt acaaatacaa acaacaatac aagtagcact
120gtggatataa acgatcccaa aaatattgca ccccctccta caacctctgt
ttcagcccca 180agcactccaa ccttgtcctc atcaagtcaa atggcgaata
tggcttcgcc tagtaccgat 240aatggtgaca atgaggagaa aaatggtggt
aagaagaaga gatttggcct attcaaaaaa 300aaaaataaat caaagaaata
atatccttcg acagcttaat gaacacgcat gtaaatgttt 360tgtaaacagt
ttgttttttc gtttttgtat ctgttaattt tctattattt atttttttag
420tttttttttg cggtagaatt gggttatcac tgaatctatt ttattttttg
ggttggtttc 480tccctgtttt cagtttgttt gaattggtgt ttaacaaaat
ctggaattat tcactataat 540tatttcctct ttttagtttg accaaataaa
ctcaactatc ttgtgttatt tctttgtatt 600atagctttac acatatttaa
ctatccatac cttttacttt ttgtatatac tgttaatttc 660ataaaacacg
ttttagtacc agaaaatcgt gtcatagcac cgttttagca gtgattttta
720aaattgtcca aggtgccaat aaatactggt gggaacaaca ttttaattag
aatagctaaa 780accttctaag aaaactatcc cagcgtagaa agaaagaaat
ctctatcagc cgcgcaagtg 840cagtatctct tatttttgca cggccccgtc
gctatattgt cacacgcatt tgaattagtg 900actcaataat aatacagatt
tgtgacgtat aaagaaacca cagaaaaact gcgtatgtcc 960gaagcacagg
tccactagta atagagccca aaaatctgac atg 100361003DNASaccharomyces
cerevisiae 6aggttgccac atacatggcc aagaccggta agtcagcctt ggaagcagaa
aaggaattgc 60ttaacggtca atccgcccaa gggataatca catgcagaga agttcacgag
tggctacaaa 120catgtgagtt gacccaagaa ttcccattat tcgaggcagt
ctaccagata gtctacaaca 180acgtccgcat ggaagaccta ccggagatga
ttgaagagct agacatcgat gacgaataga 240cactctcccc ccccctcccc
ctctgatctt tcctgttgcc tctttttccc ccaaccaatt 300tatcattata
cacaagttct acaactacta ctagtaacat tactacagtt attataattt
360tctattctct ttttctttaa gaatctatca ttaacgttaa tttctatata
tacataacta 420ccattataca cgctattatc gtttacatat cacatcaccg
ttaatgaaag atacgacacc 480ctgtacacta acacaattaa ataatcgcca
taaccttttc tgttatctat agcccttaaa 540gctgtttctt cgagcttttt
cactgcagta attctccaca tgggcccagc cactgagata 600agagcgctat
gttagtcact actgacggct ctccagtcat ttatgtgatt ttttagtgac
660tcatgtcgca tttggcccgt ttttttccgc tgtcgcaacc tatttccatt
aacggtgccg 720tatggaagag tcatttaaag gcaggagaga gagattactc
atcttcattg gatcagattg 780atgactgcgt acggcagata gtgtaatctg
agcagttgcg agacccagac tggcactgtc 840tcaatagtat attaatgggc
atacattcgt actcccttgt tcttgcccac agttctctct 900ctctttactt
cttgtatctt gtctccccat tgtgcagcga taaggaacat tgttctaata
960tacacggata caaaagaaat acacataatt gcataaaata atg
100371003DNASaccharomyces cerevisiae 7cttagtatac gattgtgttc
ttgcccacca tctactaaca aaatatggca agattaaacc 60tagtagcgca acataacaaa
ccactaataa tggagatgca cttccatcta ccaaaaatct 120tggtagagcg
ataccatgtg aagtagattg tgggccatct ggatgaccgt atttcaaata
180gttttgccta accaattcgt cagtaaggga ttcgtaagcc ttcgtaatct
gaacataagt 240ttcttccatc acacttttct catcaggtgt taggcccttt
gctaatttat ctggatgaaa 300tttaacagat aattttctat aagcagattt
gatgtctcta tcggaagcac tagtagagat 360accaaggatt tcataaggat
caaataattt tgtagcagcg tctttaatcg cgtcattact 420attaatcctt
tgcagaagaa ttgcaactaa gatccaaccc acaataatta taatatttct
480cctgctccat attttggact tcttattact atttttatca aacttccttc
taaattgttt 540gatttcatca ctggtgtatt cttcattcaa gttcttgaaa
acttcctcat taaactcctt 600actcttccct gaattcccat cttcagcatt
ggccccaaaa aaaatttggt atatttgaag 660cagtgtcata ggcccgacga
ccatcaagag ccccgttaaa atgaaggacg gccacgtctc 720actagcctca
tcatactcgt aatttgtagg cattgtgctg taatatgcag tatatttatt
780ttgttgcgaa ggccttcttt ggtcaattat aactgactat tgttccagtc
gcatctttta 840ttcgttcagt gatcttcgac gtgttcaaga caaatttttc
agaaaacgcc tttccgtaac 900gagtgaccaa ttcgcgttat aagagcgatt
gtgtcagtaa tgaagtcaga ttgtgacatt 960ataaagctaa gacgttaata
actcaaaata gaaaagaagc atg 100381003DNASaccharomyces cerevisiae
8aaataagagg tttggccaaa acgtggtcat ccaaggttga tggattcttc tcttctaata
60ttattaataa tggatcgaaa catatttgag gcaaagctag ttcccattct tttgccaaga
120aagcggaggg agaatgatcg taaagagtca caacgtccct aggaaacagt
ttttggaaca 180atttctgaaa tgtattcaaa ataggaataa tatcacttga
atagctgttt tcgtgaacgg 240atgtcgtttg accgatcgaa acagaaaagg
atttggcatt tgcagtcgtg aactgtttgt 300atgagccatt gtccaaccgt
cttatgtcat aatcggtgaa aatgctgaaa atatttgaca 360gcgaaagatc
aactgaagcc ttgatgaatg gctccaaata tataaacccg ctactttcaa
420atagtatctt tgttgtctta ttcatcatag gttttatatc tctcaatacc
tgcgggattt 480cttgcagtga agacgcactc attattatca agtcgaatac
caagccacca ttttcattat 540taaggtctac taaatctagg agcgattgga
aatggttttg aatctgaaac cgttccgtcc 600cataaaatag tgaactaacc
tcgtagttgg cattttttga gttgttcaca agataaaatt 660ctatattttt
ggcatgttga aacctggatg tgtacagaat gagatttggg tgatctccgc
720ataccaaaac ccttaatgca gatgtcatta ttcaatatac ctatcgcgag
atatgccggg 780aataaaatgt aggaaacaaa aagaaaaata tatgcacagt
agttcaaaca gaatagaaat 840gacggagagc aaatatttgg gtagaatcac
agactttagt gttggttaca agcttttcga 900atggatgtag tttgtaatca
aaggacacta ataatagcga aaaaaaaaag ggcaagtatg 960ttggcagtcg
cctctttttc aatttagcgg taagttttta atg 100391003DNASaccharomyces
cerevisiae 9cgcaaatgat gagaaatagt catctaaatt agtggaagct gaaacgcaag
gattgataat 60gtaataggat caatgaatat taacatataa aacgatgata ataatattta
tagaattgtg 120tagaattgca gattcccttt tatggattcc taaatcctcg
aggagaactt ctagtatatc 180tacataccta atattatagc cttaatcaca
atggaatccc aacaattaca tcaaaatcca 240cattctctac attactagta
tattatcata tgcggtttaa gaaaatggta taaagattaa 300gaaacagtca
tgaaaattta gtgaagctga aatgccagta ttgataatac gatagaataa
360tgaatgacaa agtatataag gaagatgaag taacattatt atggagaact
atcgactccc 420ttttgtgaat ttctatatcc tcaaggagaa ttgcttgtat
actatgtata tataatatta 480taacccttga caacaatgga ataattagtc
cttaaacgtt agactggctg ggacagcacg 540acctctgttg gctaatttta
cttttcccac aaccgtagat cttgatttca tgactgtttt 600cgtggtaatg
tccgaatgat acatatataa gttatatgct ttcactttat aagtattagc
660gtttttactc accatccact gctttgagaa gatgtggtca gcaaccacgc
aatacatcac 720atattttggc acgggtgatg taccgatcta atggctaagc
ttccttttgg aagctgatag 780agatccagat ttgcatctct atcgctattg
tcgtcatcga cactgctcac ttagtgctgc 840aggtaaattc cgtttttttc
caaattaata tttatgaacg ttatgatgtc aagttttttc 900aagaagtaat
tatccgcgaa aaaaagaata taaaaaatac aaatgtgcat agatcctcac
960atagtataca actaaaaagc aaacaaaaga acatcctcaa atg
1003101003DNASaccharomyces cerevisiae 10caatgattct gaaatactcc
ttttacaacc tttgcaaaga taatgtcttt cagtctgata 60ttacgagcga cctagaacca
ctaatccata ttcttcattc aacttactcc attttccttg 120gcaaacaatg
ccccacaatc atatacgtca taactataag ggatatgtct ggaatgcggc
180caagatagaa ttaaagggct gcagaacacc actactgata ctcattgcca
aggctaggag 240gcaccatccg tttcattttc tttgaggtaa gccaatcatg
aaatagtata cacatccata 300acggacgtac ggacgaaata agtgccgttg
tcccactatt ccaccgcatt tggcccattt 360ggctcacttt gactcaactt
gcgtcatttt aactgatatg aagggtccga ctttgtcctt 420tttcggccac
cgcatacccc acggcgatgc ctccgctacc tgcatttgag tagcatctcc
480gtttcgcggg gtattcggcg ctacgtcgcc tgttcgagcg gctctgttcg
ttgcatgaaa 540ctaaaataag cggaaagtgt ccagccatcc actacgtcag
aaagaaataa tggttgtaca 600ctgtttctcg gctatatacc gtttttggtt
ggttaatcct cgccaggtgc agctattgcg 660cttggctgct tcgcgatagt
agtaatctga gaaagtgcag atcccggtaa gggaaacact 720tttggttcac
ctttgatagg gctttcattg gggcattcgt aacaaaaagg aagtagatag
780agaaattgag aaagcttaag tgagatgttt tagcttcaat tttgtcccct
tcaacgctgc 840ttggccttag agggtcagaa ttgcagttca ggagtagtca
cactcatagt atataaacaa 900gccctttatt gattttgaat aattattttg
tatacgtgtt ctagcataca agttagaata 960aataaaaaat agaaaaatag
aacatagaaa gttttagacc atg 1003111003DNASaccharomyces cerevisiae
11cttagtatac gattgtgttc ttgcccacca tctactaaca aaatatggca agattaaacc
60tagtagcgca acataacaaa ccactaataa tggagatgca cttccatcta ccaaaaatct
120tggtagagcg ataccatgtg aagtagattg tgggccatct ggatgaccgt
atttcaaata 180gttttgccta accaattcgt cagtaaggga ttcgtaagcc
ttcgtaatct gaacataagt 240ttcttccatc acacttttct catcaggtgt
taggcccttt gctaatttat ctggatgaaa 300tttaacagat aattttctat
aagcagattt gatgtctcta tcggaagcac tagtagagat 360accaaggatt
tcataaggat caaataattt tgtagcagcg tctttaatcg cgtcattact
420attaatcctt tgcagaagaa ttgcaactaa gatccaaccc acaataatta
taatatttct 480cctgctccat attttggact tcttattact atttttatca
aacttccttc taaattgttt 540gatttcatca ctggtgtatt cttcattcaa
gttcttgaaa acttcctcat taaactcctt 600actcttccct gaattcccat
cttcagcatt ggccccaaaa aaaatttggt atatttgaag 660cagtgtcata
ggcccgacga ccatcaagag ccccgttaaa atgaaggacg gccacgtctc
720actagcctca tcatactcgt aatttgtagg cattgtgctg taatatgcag
tatatttatt 780ttgttgcgaa ggccttcttt ggtcaattat aactgactat
tgttccagtc gcatctttta 840ttcgttcagt gatcttcgac gtgttcaaga
caaatttttc agaaaacgcc tttccgtaac 900gagtgaccaa ttcgcgttat
aagagcgatt gtgtcagtaa tgaagtcaga ttgtgacatt 960ataaagctaa
gacgttaata actcaaaata gaaaagaagc atg 1003121003DNASaccharomyces
cerevisiae 12cataagaaac aaattttgta agtttcttgt gccttttgag tttgaaatca
ttgaccagaa 60caatgagaac tctaaacacg acaaatggat cttaaatcaa ttttcttgaa
aacatcttcc 120agtttcgata ttcgcacatt tgccggtaaa atagtgcaat
ctgcctatac ctttaaaatg 180ggatcttgaa cgcgtacctg aaggtttttt
taaataaggc cgaacctaaa tgttgccttc 240aaaattaacc tgtatcaaat
tttgaagccg aagttggtat cctcacattg tctaacctca 300tggtctccta
aagtccggcg atcactttcc gttgcgtgac tattgatgat cagactgtca
360acaatgtcgt tccgggcctc catagcttga agaattttcg ttgacggaac
tagaggaatg 420gcagatggag atgaccttag tttcatgtcg ccttgatagt
tatcaaagag tacagaaaca 480agacgtctct tgctcaataa taaggattgg
atcaggtgct tacaaatttt attcgtcata 540caaacaatta ctaatatttt
tttaatatta agcgtacccc tatttgattg gctctcttct 600aaattcatac
gttttaaaat ccgctttcaa ccgtcaacat aaaattagta agcagacctc
660gttccaagac ccaactaagt tgcactaaat atcctcgaac aaattggggg
cagatttttg 720aaccatataa aagtcatgac aatatttcca gatcaaggac
agttgcagtt cttcctttca 780tcggccagct gacgaattga ctgagcggct
ttgtatttca ctagaataca cttcctatag 840ctacaaaata ttattcaata
atattagtga attatttaaa cctctacctg aaccacctac 900cgagttggga
cgtctcaggg ttctttctaa aactgccggc ataagagttt caccgctaat
960tctgggagga gcttcaatcg gcgatgcatg gtcaggcttt atg
1003131003DNASaccharomyces cerevisiae 13agaacatcca aattcggaac
tactaagaag atatgggtat gttgaatggg acggttcgaa 60gtatgatttt ggagaagtgt
tacttgaaaa tattgtcgag gcgttaaaag agacttttga 120gacgaatact
gaatttttgg acaggtgtat tgatatctta cgcaataacg ccaatattca
180agaattctta gaaggtgaag aaatagtact agattcatat gattgttata
ataatggtga 240attgttgcct caactaatac ttttggtcca aatcttgaca
attctttgcc aaattccagg 300tttatgcaaa ctggacataa aagcaatgga
aaggcaagtg gagagaattg taaagaagtg 360tttacaattg atagaaggtg
cccgcgccac tacaaactgt agtgccacat ggaaacgttg 420tattatgaag
cgtctagccg attaccccat aaaaaagtgc gtttctatcg aaaaaccttc
480gaaaggaaac tcattaacaa gggaagaact aagagatgtt atggctcgga
gagttttgaa 540aagcgaaata gattcgctgc aagtttgtga agaaaccatc
gacaagaatt acaaggttat 600tcctgatgaa aagctgctaa ctaatatttt
aaagagaaag ttgacagagg aagaaaaaag 660ctctgtcaaa cgtccttgcg
tgaagaagtg agcggttgtt ctaaccacta tttaaagccg 720caattagtaa
tgcaaaaagt tggccggaat tagccgcgca agttggtggg gtcccttaat
780ccgaaaaagg acggctttaa caaatataaa ctccgaaaat ccccacagtg
acagaattgg 840agaaacaacc agttttgata tcgccataca tataaagaga
tgtagaaagc attcttcact 900gtaatgtcca aatcgtacat ttgaatttct
tgtaggttta tttaaaaggt aagttaaata 960aatataatag tacttacaaa
taaatttgga accctagaag atg 1003141003DNASaccharomyces cerevisiae
14gttgaacttt attcaaatat catcgaagta agaagatata ctactaaaga ctccctttgc
60tctatattcg agtcaggctc caccagtcat ttcgaaatta accagttaca ggtcaaaaga
120ctaaacttac tgcaaaacca atttgcttct gttttcactt ctttccaccc
caaagataat 180acaaagggca tacatatcaa ttttttttca cctgtcacta
ggataactga ccttcaatat 240tccttttttt atacgaatca gatactgttc
ggtacacgat atctaattaa aatgattcaa 300aactttgtaa caggtaaagt
tttcactaga accaatcaat ccaagtgaat taggggaaac 360catagttgtt
gatttgtaga aacctcacat tgtacattgt tggtttgttg ggcatatcag
420aacgagagat tttccaacat tcaatataca ctaaacccta tgacgagtcc
cacagatggc 480gtaaggtttt tatgatttca gcagggtacg acgactagta
ccatattaac attttttagt 540gtttctaatt tgggaaaagg tccgtgtttt
ttctcctagc aaccgtttag tgccaagggt 600taggcaattg aacgaggcca
agacaatatt ggctttgctt ctattacttg gctaacattg 660tgtctgcagg
tcgaaaggca cctttactgt aaggaacatt cttgcgctct aaacatacga
720agatatgggg aatatgaagc gtgtttctta tacgaagtgc agcatcgttc
aaggaaaata 780cacccccata gtaataatgg ctaagtggcc aggaattaga
atatgtgaga tatgagtgca 840aaatgagtga ccagtaatag cctgtttggg
atgtaattgc tcaaaaaatt tatataaata 900cagcggtttg atcagctttg
tttgagacat ttctctgttc ttttccttcc agttaagctt 960atatctccac
taagcaacaa cccaaaaaac aacaaataca atg 1003151003DNASaccharomyces
cerevisiae 15cagacttgtt gtctgtgctg ttacttgcac ttgctgcatt gctatttgcg
ttcaaatcac 60ctttttcgat gaggaggtcc ctttgcttct ccaccatcgc ctttgtgtaa
cgcaacattg 120tccaatcgaa caagtggtcg ttgtgatact ctagtttaat
actcagatct ttaaacagcc 180ttgccaagaa caaataatct ggcttctcat
cgaatttcaa attcttacag taagccatat 240attcttgaaa ctctaatggt
aaacctgaac atagagtttc cacgctaacg tttaatttct 300tttccatgat
acgatcatac ttttgtttct
tggtggttgc tttcaaaccc tgccatggca 360aagaaccctt acaaaaatag
atcaagacat aacctagtga ttctaagtca tctcttctac 420tttgctctat
tccaagatgc gtattgacac ttgcataacg agctgtacct gtcaaggact
480tgttctccct gtaaggaata tgacgatgtg tgttgaaatc tcggtatttc
tttgatagac 540cgaaatcaat aacatgaacg gtgctaccac ggcgtcctac
ccccattaaa aagttgtctg 600gtttgatatc tctatgaatg aacgaccttc
catgtatata ctgaatacgg caaaacattt 660gcaaagccag catgataacc
gtcttaaagg agaaccttct gtgacagtag ttgaataaat 720cttccaaaga
tgggcctaga agatcgatga ccatagcatt atattcaccc tctctgccaa
780accatctgat gaacgggatt cccacaccac cgcttaagta tctgtagacg
cgggactcat 840agtccaattg aggatgtctg gacctgatcg attccagctt
gatggctact tcttcaccac 900taattaagtt cgtgccgtgg taaatgtcac
caaaggaacc actcccaatc ttcctgccaa 960tacgaaattt ccttcctact
cttaagtcca tctcttttta atg 1003161003DNASaccharomyces cerevisiae
16tcactcaacc acctccaaaa aataacaggt tcatctaaag taaaagactt taacttgctc
60ttagtttcca aattaaatat ctgcacgata gtaccatttg ctctaacgga aataaccatc
120tgagatggat gcatgatagc agaatcaccg cccatattct tccttgtcac
ttcattgcct 180ttggccaaat ccacgattgc aacagagttt gtaccgtcct
ttgtttctct aacagtgacg 240aagtggtcac tctcgaaagt agttgatctg
aagtcaagga attgagggga aattcctaag 300gacatcagat cgaccaattc
ggtaaattca atgggtaggt cactcattgg ttagaacttt 360cgtgataatt
tatttttata gttgaatatc ttctttctct ctcaactctg atccggattg
420tcgaggtttc aataagttac tctgaacaac taatcaaaat atctccttat
ttctgtagat 480tccttcagtt ccacttttta cttttcttaa ttctctttgt
atttattcct agcgacgaaa 540aatgcgagat ctcgaccaaa aaaagggggt
agggtaataa aattaaccct attatttttt 600aactttaaaa cctataatgt
gctaatattt tattataaac ctcctttttt tgcgttcaaa 660ccctgacaca
ttttaagccc tatatttacg gtattagttg attaaactcc gaagcgaaag
720gaattcggtc attagcggct aatagccgtt ggggtaaatc acctacaagc
aagtacacaa 780gagaacgttg gcgttgttaa gtcaaagcac taatacattg
gggctttaag agtgtttata 840aaggtctaac ctgtaaaaat tatttaaaca
acttgaacag gccttaaagt tttcctcatt 900ccgctcatca tcactaatat
tgctctccgt ttttgaatac acacttgaca ctaataagta 960tcacagaaaa
aaagaaaata taataaatta gtattgcgat atg 1003171003DNASaccharomyces
cerevisiae 17aaaacggctc ctagttatcg tcagttttga taattagtaa ctagaatata
gtatcaccaa 60gaacaggcca ggtaatatac tagaagttgt gtataactac taatattttt
atacatatag 120atgccaagtc aaggtccaat tagtagattt ttcactctac
cttgcattat tacctgaaag 180atcgtttgtg tttccttaaa ttcgccagag
gtgcaaacga atgtaaatat ctacatcaat 240ttacggtttc acgcccgttt
cgtttctaat ggcaactcat gtaaactgtg ataaacccgt 300tatttcagtt
ttattttctg ctagtataaa ggatgatagt agccatttcg ctggaatatg
360tatcgggttt aaaagatggc atagttcgaa acattttcgc acaagattct
aactgtcttc 420gaacgtcgta gaaaccgcaa cgcaagaaat gacagcgcaa
taaaatttat tttactaact 480gatagaacaa atgtaaaaaa tttcataata
tcaagtagaa ttctttattc tctttgtgaa 540ttcctaaacc catgaaaggg
acttgtagta taagctggat acctattttc attggcttcg 600tttccaaatg
acacaaaacc atcacaaaag gaaatagtcc ttccatagtt acccaatcat
660tctgttgcat attaaatgaa cagctccttg ttgataatgc cagtgtggca
tattcaccgc 720tgcagttatt tcttttcata tataatgagt catttgtttc
tgtcatcact tttctatact 780ttctcttccc cgcgtgtttt ccgtacacca
acaatatatg ccataataca cgtaacattt 840ttttataaaa agaaaaggta
agtgatatat ataaaatagc gccatgagta ggaaactttt 900ctgttactgc
agatatgtgc cagactggct caggtggcat aaacacagat taatagtatt
960ggcgttgctg aaataagaag atctgtaaca atatcttaca atg
1003181003DNASaccharomyces cerevisiae 18tctaaaaatg ttcaggacta
catacatcaa ttaggtttca ttccaaaagt accttttgtc 60aatttatacc caaatgccaa
ttcacaagca ttagacttat tggagcaaat gctcgcgttt 120gaccctcaaa
agagaattac cgtggatgag gccctggagc atccttactt gtctatatgg
180catgatccag ctgacgaacc tgtgtgtagt gaaaaattcg aatttagttt
tgaatcggtt 240aatgatatgg aggacttaaa acaaatggtt atacaagaag
tgcaagattt caggctgttt 300gtgagacaac cgctattaga agagcaaagg
caattacaat tacagcagca gcaacagcag 360cagcaacagc aacagcaaca
gcaacagcag ccttcagatg tggataatgg caacgccgca 420gcgagtgaag
aaaattatcc aaaacagatg gccacgtcta attctgttgc gccacaacaa
480gaatcatttg gtattcactc ccaaaatttg ccaaggcatg atgcagattt
cccacctcga 540cctcaagaga gtatgatgga gatgagacct gccactggaa
ataccgcaga tattccgcct 600cagaatgata acggcacgct tctagacctt
gaaaaagagc tggagtttgg attagataga 660aaatattttt aggacaaaaa
actataagta accggggaag tatagaatca ccatagatgt 720aagcttacag
acaatgtgta tatatgatgt atatgaacgt atacaaatat atatatatat
780acgtgctctt gttgtagctc gtatatcaaa ttcctcctcc gacgcttatc
ttaatcgtac 840tccgcggaag tttgttatcg cctcttgaat tctttctttt
cgttcattta tgattagtca 900tctatagaca atattcatta tttaagcacc
tagaatacta aactaaatgt ctaaatatga 960cacaaggaag ataagataaa
aaaaaccaag cgcttagaat atg 1003191003DNASaccharomyces cerevisiae
19catcgggatc ttccgactcg tcatcagacg tcgaggtcat tatgcactcc ccgagcgatc
60ctgaatacgc tttaaaatca caaactctaa gaagctcgag tcagaccgtc attaatagta
120agagaccagt aaagatagaa gacgaggaag aagccgtagg aatgtcacag
ctcaattata 180ggtcgtcatt aagacaagct cctccaagag ctccctcaac
tttgtcatat aatcactcga 240agaacaacga aacgccaatg caagatattt
tcacaaatgg cgaaacagca aataacagaa 300agaagaagag aggatctttt
gcaaggcata gaacgatacc aggatctgac gtcatggccc 360aataccttgc
acaagtgcaa cattcgacat ttatgtatgc agccaatatt ttgggcgcct
420ctgcggaaga caacacgcat cctgacgagt agtatagctg tgctgagcct
gaagtataat 480gcatatccat cggactattt agacaaagcc caaggaagcc
taaggcggcc tgccaaggtg 540tttctccctt tttttctccg atttctttgt
ataaactaaa taatatagtg attactaatg 600gaatggcggt attatgcacc
taacctgttc attctgccac agattacgta agcgatttat 660tgccggcact
tgttgtctta atgaccgagt caccaatgtg gaacgataat tttctctgac
720tcaaacctgt taattttttt ctacttcgtt tcgttagcga cgacgtcaag
ccgcaggatc 780ctgtctgcct tgacctcttc tctaccttca gaacttacac
ttttcaggta agatgacctt 840tatataaaag ttgacactat tacagttgtt
tataaacgtc tgaagaatga gacgttttta 900taaaatgaat aaaatgcata
ttctaagttt aaaacaacat tttcaaagtg tacgattgta 960aaaagagagg
caattagaga atctcaaaca ggtaataata atg 1003201003DNASaccharomyces
cerevisiae 20gaaatccaat gagaataccg cataatcttt cccaattacc ttgatattta
caaaagacca 60tagatcctta ccctggtaaa gcttgtaatt ctgtgaatcc gttccctcca
attggatttg 120attagtggaa gtcaacttgg acagtaatgt actacccggg
tttttatgca ctccatagat 180caaaggatat ggaaaccaaa attccgtcga
caattgtggt gactcaaaga tcaaatggtg 240agtagtcaaa tgtaaagtac
cttgtgtggc cgtacctctt ctatgcagca ctacattaga 300tactttggca
atcttgatgt actccatctt tgcggaatgg tatactattt cttttccctc
360tttttagcac tattacaccc cgcccacaaa ataaaaataa taacgacgac
ctaatctcac 420caagtgaccc ttgtaaaacc tccttttctt tataatgttt
cttttcttac taatatttgg 480tacatttagg gtagtgataa aagaatggca
acattgttat tattgtgaaa aatgaggaaa 540atagaaaatc agaaacccta
aaaagtgatt ttaccctatc agaaatattc aaatgtccta 600attaaaaata
gtaaatcccc taaacattca gattgtaaac tagggttgag aaaatgactc
660atccaccact gtcttctttc ctgcggcatt ctatagatta ttgtgaatga
ctcttattga 720tgagatggca ataacttttg aatatcagag ataggaacct
ccatgtcgta acgattgtgt 780caccttgagt aagcatcgag aaaatccaat
cttttttttt ccgtcataag catttctgcc 840atgctatttg tatatatata
attactaata cgtcttctat agtatgcctt atctcttttt 900tgaagcgcta
tttaagttta agcatcgaaa aactaacatc tatagttaaa attagttcta
960taaaggaaga gcaatacagt acatagacag gaagaaaaga atg
1003211003DNASaccharomyces cerevisiae 21gaattacgag aagcccaatt
gaccatcgaa aagctacaaa ggaaacaact acactacaaa 60aggctactcg atgaccaaag
aatggtcctc gaagaagtgc aaccgacttt tgataggtat 120gaagccacaa
tacaagaaag agagaaagag atagaccatc tcaagcaaca attggagctc
180gaacgcagac agcaagccaa acaaaagcag ttttttgacg ctgagaatga
acagctactt 240gctgtcgtaa gccaactaca cgaagagatc aaagaaaacg
aagagagaaa tctttctcat 300aatcaaccca ctggtgccaa cgaagatgtc
gaactcctga aaaaacagct ggaacaatta 360cgcaacatag aagaccaatt
tgagttacac aagacaaagt gggctaaaga acgcgaacaa 420ttgaaaatgc
ataacgattc gctcagtaaa gaataccaaa atttgagcaa ggaactattt
480ttgacaaaac cacaagattc ctcatcggaa gaggtggcat ccttaacgaa
aaaacttgaa 540gaggctaatg aaaaaatcaa acagttggaa caggctcaag
cacaaacagc cgtggaatcg 600ttgccaattt tcgacccccc tgcaccagtc
gataccacgg caggaagaca acagtggtgt 660gagcattgcg atacgatggg
tcataataca gcagaatgcc cccatcacaa tcctgacaac 720cagcagttct
tctaggcagt cgaactgact ctaatagtga ctccggtaaa ttagttaatt
780aattgctaaa cccatgcaca gtgactcacg tttttttatc agtcattcga
tatagaaggt 840aagaaaagga tatgactatg aacagtagta tactgtgtat
ataatagata tggaacgtta 900tattcacctc cgatgtgtgt tgtacataca
taaaaatatc atagcacaac tgcgctgtgt 960aatagtaata caatagttta
caaaattttt tttctgaata atg 1003221003DNASaccharomyces cerevisiae
22tgatgcagtt ggagtcggta gcgatgttat aggaggagga ggtgcactag gaggcaaagg
60aggcgccata gcatccgtat gggggggtgg tgcgttaaaa gcattcgatg atgtttgctt
120atgtaatttg gattcaattt catccagaaa cgacatacca ttttggctac
tctgtttagg 180cgggtttata gatggagaaa taggagaagg tcttacgtta
tcatccttag atgaacctat 240tacatcctct tgtttatctt gggtagtgta
tccagtatca ccaccaacca cgaatctatc 300atctctcttc ttttggatct
cagcaagaaa tggtaagggt cccccaggtg ttacggccga 360ggaagaactg
gatgaagata ttttggattg cttactggac gctgagggtt tgtttgtcgt
420gagtgttgga ggaggaggag gagccgatgg tactgacgtt gcaaccgaag
gaggaggagg 480aggtggtagt ggtgctaaag gaatagatga tgctgacaag
gcagaggatg ttgaaaaagc 540accaggtggt gggggaggag gaggtggtgg
cgcgggcatt gacgatgctt tgttaggagg 600aactgatgta gtatttggta
acggtggtgc taaagttggg ggaaccgggg ttgcttttac 660gctgttagtc
gacgcagagc tcatggctgc tggtagtggt ggtggaggcg gagctggagc
720tgaagttgct tttttaggtg cagaggtcac atttggtaac gatggtggag
gtggtgcagt 780aggcggtaat ggcggagcat gtgatgtagg aatgggcgga
gcagatgatg gaagtggtgg 840ttgtgaagga gatttatgat tttcggtttg
aatctttgtg gaagaaacgc cctcgacggc 900tcctcgttca gacctcctgg
cattaatttc tgccaaaaat ggcaacccgc ctgcaggaac 960ttcagaagat
ggtggtgaga ctgcgttatc tgtaggttta atg 1003231003DNASaccharomyces
cerevisiae 23tgaataagcc tgtataactg cgcatgtaag aatagtggct gcaatagatc
ccacagcagt 60cagtgctcca gcctgctctt tcaaatccgt cgtcgtaagt ctatcaacag
gtaatcggtc 120aaatgtgaca ttatttgaga ttcccaacca aaatgtgggg
taccaaagta ttacttttgc 180tatgggcgat ccattttcta gtgttctttt
caccgcactc ttccaaacga cgtataaaac 240aaagcataaa ggtaaaagtc
tttgaaatcc cacactgaaa agctcataca cgctagctgg 300gtcatgtgca
tccgttagta atggagggtc tagagataat acaagagtcc aatagcccac
360tatcgtgttt atccagtacc cgtaaatact gtcatacaaa taaaatattg
gcagtccaaa 420aagcatattc aatgcgacca cagctgctcg aggatcataa
cagcccgaca ggataccgcc 480ttgtatatct ctgaatgaat atgaaccggg
aaaaaatgaa ttgaatgcta ctgaaaaccc 540agtgttatac attccttttt
ttgatggaaa agaattttgg gccccttgca tagaaacttt 600ggtacagcca
ccgtaaaatg gtgagataac acctgcatgt actgcagagg cgcatggaaa
660cgagtcactt ctataagggt aggacacgac catcgggtct tcttcgctaa
acagggcacc 720tccgccaatt tcataccctg tatatttgac tctcctattc
cctacggcaa tggcggagta 780tgtccatcca cctctatcac acagtgctgg
acatctgatc atgtattctt tattatcaag 840tggaccgcag tttttagcat
ttaaaccaca ttcgttattg gtaccctccc agttcaagta 900ggaattgcaa
cttaaagaca atataggaat cttttctgag ccatcatttg gatggaagta
960tggcggcttg ataagatagg ggtatatgag agaataaaaa atg
1003241003DNASaccharomyces cerevisiae 24ctggctattt atataagatg
ttgctctaat tccatgagtg atgtcccatc gcatgatgaa 60gaagttatga atgaagtgaa
caatatcatc gaattacaac aacggcccat gcaaatgacc 120aagtctaccg
tacgtaccag gagaagacca ccaccacctc ccattccttc cactcaaaag
180ccatcgttga cagaagagca aaccgaaagt ataagaatgt cacgccgtaa
taaagatgag 240aacaatgcta aaagagttgc cccaccaccg ttgccgaaca
gacagttacc taacttagac 300gctaatactt actatgtacc atcctcacaa
aatgactatg gcatgtatgg tgctttcatg 360gataaaaaag cagatgaatg
gaagagaaga gtaatggact caattcaaaa attgagcaat 420caagacacca
cattgatgtt cttttcggat ccggcattaa gtttggaaga cagtattcgc
480agaattaggg agaagtattc aaactaaata cactgttatt tatatacatc
gggagggagt 540ttaaaaattg tatacactaa agtgacctta tgaattggag
ccattttcaa cgttttagag 600tgatgttgca tccttttcat cagatcggga
aattaaaaaa acggttatag aagggctaaa 660tgtaaatatg taaattttat
gaagtcaaga gtaaagaaaa tcttcggttg ccgatcaaat 720acactcttac
aaaataccct aaactcagtt tatctttcca ttccaactga ttatcgcgta
780cgccttggtg accaaccata ccctaaaatg tcaatttccg gggcggaagg
ggtccttaaa 840ctgcaaagcc tcttcccatg taaataaaat aaatccgtag
aatttattaa atattgtcat 900attataagtc gggaaaggcc ttctgaatgc
acgtatattg accaaataga gttagattca 960aattatttat tacatttttt
cagcatcgaa gttaagtaga atg 1003251003DNASaccharomyces cerevisiae
25ggcgaaagag ctgtgtatga gaataacgag accggtgtca ttaagaacct ggaattgcaa
60aaggccacca tccagggtta cgacaatgac atgaggccag tcatccttgt gagacctaga
120ttacatcatt cttccgacca aactgaacag gagctggaaa aattctccct
gttggtcatc 180gagcaatcaa agcttttctt caaagaaaac taccccgcat
ccaccaccat cttgttcgat 240ctgaacgggt tctccatgtc caatatggac
tatgcccccg tcaagtttct aatcacttgt 300tttgaggctc actacccgga
atctttgggc catctactta tccataaggc cccttggatc 360tttaacccga
tttggaacat catcaagaat tggctggatc cggtcgtcgc gtccaagatt
420gttttcacta agaatatcga cgaattgcac aagtttatcc aaccccaata
catccctagg 480tatctgggcg gagaaaatga caatgatttg gatcattaca
ctccaccaga tggctctttg 540gacgtccatc tgaaagacac cgagactcgc
gctatgatcg aaaaggagag agaagaattg 600gtcgaacaat tcttgactgt
cactgcccaa tggattgaac accagccatt gaacgatcca 660gcatacattc
aactgcagga gaaaagagta caactttcca ccgctctttg tgaaaactac
720tccaagttag atccgtacat cagatcaaga tccgtttatg actacaatgg
ttctctaaaa 780gtttgagcca agccacgcga cgctccatac tgtatagacc
tttattataa aaacatctat 840ttctttatat gtccctctat cttatttttc
ttttactttt atttgtataa tacgcgaaaa 900attatagaat catttattat
caaaaacaat caactatgaa atgaaaaaca atacgcctgc 960taatacggtt
cgcccttcaa tttttgtttt ttttacatga atg 1003261003DNASaccharomyces
cerevisiae 26agataagaat atgagggtaa caatgaatga gaactcactt tgtaatcacc
agttgaagaa 60tggtaaacaa ggctagtaat acatcctggt atttcggtct catgttcaaa
ataatcagtg 120gtaatttata ttatgtatat cttttccttg tttctttgtt
aagcttttgt gtaaacatac 180tttagtaatt gtgacgtttt ttggcacttc
gcacaggcgt gactagtttc tgtgaacaga 240gggcgtctat tcatgatctt
cacaaatctt gggggagtta atttactctg cttccacatt 300acattgatgc
gcagtgatct actgacgcct gctggtggtg agtacacatt agataaatac
360ctagggtgtg cagcaataga ccttaaacaa gggtgagctt tgcctattag
agaagaacct 420ctaactgcgc ttttcaataa tagccaccga acaattggca
tattatgcaa cctgtgtttt 480taagtacctt tttatgtcag attttttgaa
ctatccttcg attgttattt cttgatataa 540cgtttacatg atttcttaat
ttacaacgcg gacgaattag attgaaatta tttagcatat 600ctgtctaaga
gcacattaga tcgaagtacc gtagcacatc tgccatacta tcaaatgcat
660catcacgaaa aggcgcagtg ttgatggata gtaaatatga aaaggatctc
ttcttaggtg 720ttccaagcct cgcttcacgt gtaacataca cagtttagaa
gtttaccaaa ccgcggcaag 780gcgctgtaag gttcaagagg ctttgaatta
tgctatctta ctatcttgcc gcaagttgct 840gtcaggtgta gttcctaggc
ggctcgttga caagaccctt ctgcccaatg acttctatat 900aaagtatggt
tttcttttta aatacagtaa ttccatgaaa aaaagagggc caaaaagatc
960aagcaataaa ccaaccgata tataaaacac agaactgcag atg
1003271003DNASaccharomyces cerevisiae 27acagaagtaa gtaccggggt
acctaaatat acgcataaaa gtctctttct tttttttttt 60ttttttttta gcttcctaca
tttcgttaat aatattatat agattatatt tatatttaag 120aaaagtaata
tcagcatatt atgaatagac aaaaaagtct aaggtcaaga tttattaaat
180gttagattat taagattaca attaacaaca tttgttgtta cttcataaaa
tttcgttact 240tcataaaaga actacagtcc cccacttcta aatgagtggt
tgtaggggtt catatacctc 300ccatactgtt ggaataaaaa tcaactatca
tctactaact agtatttacg ttactagtat 360attatcatat acggtgttag
aagatgacgc aaatgatgag aaatagtcat ctaaattagt 420ggaagctgaa
acgcaaggat tgataatgta ataggatcaa tgaatattaa catataaaat
480gatgataata atatttatag aattgtgtag aattgcagat tcccttttat
ggattcctaa 540atcctcgagg agaacttcta gtacattcta catacctaat
attattgcct tattaaaaat 600ggaatcccaa caattatctc aaaattcacc
aactcttcac atacatacct gcggtagcaa 660gataggtaca ctttttctac
gtttcacgaa ggtagcgata ggtacctcac tcattgtagg 720tgcgggggta
gcgatggagg tttctgtacc attaccacca cagccactat actcacgtag
780tgaagtccct agcgtggaat tgtgtggtat cgttgctatc tgtcgttcgc
caccctcggt 840atatccaacg tgcaggccaa tatcactctc taagaaaatc
gtatcaggat tggtacggac 900aaactcttca tagcctacgt cgaaatcaaa
ttctgctaaa gctgtgactg tagttaaacc 960taaaaccgcg attattgctt
gaaaaactgt ggagaaattc atg 1003281003DNASaccharomyces cerevisiae
28tgtcatcgcc gccagaacgt gctccttact gtgggacgtg ccactggtgg gagtaaacca
60ctgcattggt cacatcgaaa tggggagaga aatcactaaa gctcaaaatc ctgtggtact
120gtatgtaagt ggtggaaata cacaagttat tgcatactcg gaaaaaaggt
accgtatctt 180tggtgaaacg cttgatattg ctatcggtaa ttgtcttgat
agatttgcaa gaactctgaa 240gatacctaat gagccctcgc ctggctacaa
catcgagcag ttagctaaaa aagcccctca 300caaagaaaac ttggtagaac
ttccctatac agtaaagggg atggatcttt cgatgagtgg 360tatattggct
tccatcgatt tacttgccaa ggatctattt aagggcaata agaaaaataa
420gatcctattc gacaagacaa cgggcgagca aaaagtcact gtagaggatc
tttgctactc 480tctgcaagag aacctattcg ccatgctagt tgaaataaca
gaaagagcta tggctcacgt 540taactccaat caagttttga tcgtaggcgg
tgttggttgt aacgtgcgat tacaagaaat 600gatggcgcaa atgtgtaaag
acagggccaa tgggcaagta catgctacag ataataggtt 660ttgtatcgat
aacggagtta tgattgccca agcaggacta ctagagtata gaatgggtgg
720gatcgtgaag gacttttctg aaactgttgt tacgcagaaa ttcagaaccg
atgaagtata 780cgcagcctgg cgtgattaac gatttttgtt acagttagcc
taatgttcac actacgaata 840tatatatata tttatatatg ccatacacct
tacatttaca gctcagtgta taaccttgtg 900gaagcgcgaa aataagcaag
agacatttga aaataatcaa taaacaaagc tcaaaaaaat 960tagatcatca
cctaacaaaa taagacgtgg caattacaga atg 1003291003DNASaccharomyces
cerevisiae 29ggcaacatag cagtacgtcg ccaaaaagaa cgaaaacaaa atagatatta
tgcataaact 60aacgtgattt tcatacagat attgcaattc aggcaattgt ccatcagtca
atttccatct 120gatagccaaa actaagacca aagttgtaga catggcaatt
ccattgatct tatacgaaag 180cttcgaacca tcccttaact gaacaccctt
catgactctg cccggtaaaa tgacgtccaa 240aactgccagt attccatacc
acaggcaata gacagtccat aattcacgat tgcccagata 300gtagcgcaat
ggcttgatac cgttccaaag ctcaactata tcgaaattct ggaaaaatcc
360cttaataaaa taatcgggcc ttatcatttg attcaagatg atagtgaaaa
caggcagccc 420tatgctgatg cctaaggcac caatcagccc accaaattca
aactctgtag ttctgggatt 480caaagctgat accatccttt tattctctac
accgaatttg tctttactcc tatgctgttt 540acaaggtcta tctgataagc
aattgcgcaa gaaaatagta gaatgaaaac tgattattaa 600aaacaaacgt
aaactcaagc ctcacttgat gctcagacgg agtacgtgaa aaacgtccgt
660tatgcaaaac cctttatatg cacaaccttc
acacaatgca aatttccgat gatgcctaca 720tacaaaagag cgaaaggcga
tataaatttt tttcacggga ttttcgttta ggtgaaaata 780aaatgaacga
cagagcatgc agagtccggg taatacatat gtttcaatac tgtttcaata
840ctgtttcaga agtgcgtcac atattaattt taacttataa ctggcctgtt
gctggcaaga 900ggtatatata tatgacgaat gtgaccaaca taagtcctta
agataatccc gaaatatttg 960gttaggatga ttccctttcg aatttgtgaa
cgttgatgat atg 1003301003DNASaccharomyces cerevisiae 30ctagcggtct
acattcgcta tcaactgaac gaatagtgat gcaatctgaa ccttaaactt 60ttcgctcgct
tttactttcc agaaaaaatt ttcttcattg ggattcctct gctgacaaac
120gggctcacag aactgaggaa acgagaattt tgttaatcaa agcaacatta
cgggaaggaa 180cggaaccgtc tcgttgctgt gggcgcgagt tcgacgcaag
aaaaaaatgc tgcagcatcg 240tcttttgtga acgccaagaa ggagtccggc
gaaggtattg ggcgtcacaa ccttttattc 300tccgcagtgt ctcttgccta
caaagcctgc agaagctttt atgccggggt gtacgtatgt 360taatcagtta
ccctttccta ctgtgtgccg cacccctcat ttgctctcga tacgagaagt
420tccgcgccga ttctggcctt tttgttttct ggctagccta ctttcagggg
acgtgccttc 480ttatgccgct gtaaattaca gcagagattt agatctgagt
gaaaacgtgg cgaggaaaaa 540gggtacaccg ttaaaagcgt agacacaatc
ccactttgtt gtgcctagtc ttctctgctg 600tattttaatt ttctgtttga
acttttttcc gttcttttcc atcattgttt aatttttact 660ttgagtgggc
ttctcaaaag aaaaaaatag ctggtattgt atttttaccg taggcgtttt
720ctttgtattt ttcccaagag cagcagcacc ttttatatac cttggttaca
cattgtaagc 780ctcaaaaatc gtcctatgta agtgtcttat cgcatcatac
ttaaggtcaa acctgtaaag 840cgcgcttcta gttttatttc tactactcgc
cttggctctc gagatcgctt aggtagttta 900atagttagtc acacccaccc
acccattcgc ttagaatata gaatatagag cagttgcagt 960acacgactca
ttttagcaac gataagagta taaattggaa atg 1003311003DNASaccharomyces
cerevisiae 31tccttataac ctgtaaaaaa aaaaaaaaaa aaaaaaaaga agggtttaaa
taaaaatcgg 60acttactcaa agggttgaaa agcactttaa tataggtttt tagtttcggg
taagaagatg 120tgtcaaaggt ctcgaaaagg aaacattagg gcaaatacgt
aaaggttgcc ttcgtatctc 180gtttccacag caacgagaaa cgagatacga
aggcaactga ttcccagcat ataaaaggcc 240atgaattttt ggcgtttcct
attgtctttt cgaaactatt tctttcattc ctttcctttc 300ttgttctttt
acttataaga aaaataatat acgtatatat ataactacaa accacatcag
360caataaaaaa aaactatatg accatggacc aaggccttaa cccaaagcaa
ttcttccttg 420acgatgtcgt cctacaagac actttgtgct caatgagcaa
ccgtgtcaac aagagtgtca 480agaccggcta cttattcccc aaggatcacg
ttccttctgc caacatcatt gccgtcgaac 540gtcgcggcgg tctttctgac
attggtaaga atacttccaa ctaagagcat gcttctcttt 600ttttttgtag
gccaatgata ggaaagaaca atagattata aatacgtcag aatatagtag
660atatgttttt atgtttagac ctcgtacata ggaataattg acgttttttt
ttggccaaca 720tttgaaattt ttttttgtta cctcgcgctg agcccaaacg
ggctccacta cccgccgcgg 780tcgccatttt gggaagtcat ccgtcccaaa
aaggaaatag ccataacata tcgttactgt 840tttggaacat cgcccgtttc
gcccgattcc gcctcagcgg gtataaaaag agatcttttt 900ttttcctggc
tgtcccttcc catttttaaa tgtcttatct gctcctttgt gatcttacgg
960tctcactaac ctctcttcaa ctgctcaata atttcccgct atg
1003321003DNASaccharomyces cerevisiae 32aatggcaaac tgagcacaac
aataccagtc cggatcaact ggcaccatct ctcccgtagt 60ctcatctaat ttttcttccg
gatgaggttc cagatatacc gcaacacctt tattatggtt 120tccctgaggg
aataatagaa tgtcccattc gaaatcacca attctaaacc tgggcgaatt
180gtatttcggg tttgttaact cgttccagtc aggaatgttc cacgtgaagc
tatcttccag 240caaagtctcc acttcttcat caaattgtgg gagaatactc
ccaatgctct tatctatggg 300acttccggga aacacagtac cgatacttcc
caattcgtct tcagagctca ttgtttgttt 360gaagagacta atcaaagaat
cgttttctca aaaaaattaa tatcttaact gatagtttga 420tcaaaggggc
aaaacgtagg ggcaaacaaa cggaaaaatc gtttctcaaa ttttctgatg
480ccaagaactc taaccagtct tatctaaaaa ttgccttatg atccgtctct
ccggttacag 540cctgtgtaac tgattaatcc tgcctttcta atcaccattc
taatgtttta attaagggat 600tttgtcttca ttaacggctt tcgctcataa
aaatgttatg acgttttgcc cgcaggcggg 660aaaccatcca cttcacgaga
ctgatctcct ctgccggaac accgggcatc tccaacttat 720aagttggaga
aataagagaa tttcagattg agagaatgaa aaaaaaaaaa aaaaaaaagg
780cagaggagag catagaaatg gggttcactt tttggtaaag ctatagcatg
cctatcacat 840ataaatagag tgccagtagc gacttttttc acactcgaaa
tactcttact actgctctct 900tgttgttttt atcacttctt gtttcttctt
ggtaaataga atatcaagct acaaaaagca 960tacaatcaac tatcaactat
taactatatc gtaatacaca atg 1003331003DNASaccharomyces cerevisiae
33ttcttttttt tttttttttt ttctttttgg gaacggggga aaaggtctca ctagtatata
60aaggaaagag agtacagcac gaatgagcca gctgcaggaa aggggctagg ttaaaaaata
120aaaacccagc agaaagcaaa caaccaaata taatttagaa atggacagag
accatattaa 180tgaccatgac catcgaatga gctattccat caacaaggac
gacttgttgt taatggtttt 240ggcggttttc attcccccag tggccgtctg
gaagcgtaag ggtatgttca acagggatac 300actattgaac ttacttctct
tcctactgtt attcttccca gcaatcattc acgcttgcta 360cgttgtatat
gaaacgagta gtgaacgttc gtacgatctt tcacgcagac atgcgactgc
420gcccgccgta gaccgtgacc tggaagctca ccctgcagag gaatctcaag
cacagcctcc 480agcatatgat gaagacgatg aggccggtgc cgatgtgccc
ttgatggaca acaaacaaca 540gctctcttcc ggccgtactt agtgatcgga
acgagctctt tatcaccgta gttctaaata 600acacatagag taaattattg
cctttttctt cgttcctttt gttcttcacg tcctttttat 660gaaatacgtg
ccggtgttcc ggggttggat gcggaatcga aagtgttgaa tgtgaaatat
720gcggaggcca agtatgcgct tcggcggcta aatgcggcat gtgaaaagta
ttgtctattt 780tatcttcatc cttctttccc agaatattga acttatttaa
ttcacatgga gcagagaaag 840cgcacctctg cgttggcggc aatgttaatt
tgagacgtat ataaattgga gctttcgtca 900cctttttttg gcttgttctg
ttgtcgggtt cctaatgtta gttttatcct tgatttattc 960tgtttcattc
cctttttttt ccagtgaaaa agaagtaaca atg 1003341003DNASaccharomyces
cerevisiae 34cgctcccacg acatgcgcat ctcttccaaa aacactacca ctttctactg
actcactggc 60catttgagaa ccacagagac ttggtagatt ccatcgctgt aaacaatggg
aaactaaatt 120caacgtcttc aaggagcgtc tggctgaagg cggattggat
aacactgttt aacgttaaaa 180acccttgggt ccaaacgccg ccatcgttaa
tgcgtctgag tgggacagat cttgatacct 240tcacaccaga gaggatattc
ctgataaatt ctcttggaaa ccactacaaa tttttaatag 300cgaacagtca
tctaagctac aatcacaaaa aatacccctc ccctggcgtg caaattcctg
360tcaggaacgc tcttggcgaa gtttctccgg ccaaacaaat tgcccaactt
ttcgcaagac 420agctgtctca tatttacaag agcctcttca tagaaaaccc
cccgctctct cctgagaacg 480agctggcatt gactgccgtc ttctatgacg
agactgtaga gcgacgcctt agaaggctct 540atatgcgagc ttgtgcaagg
gcatatacga ctactaatgc cgactccacc acggagcctt 600tgatgtttca
ttgcacccgg tgggaagttg attagttgtt gttggctgtt atactggagt
660aaatgtcacg tgccactgtg gccgtgtcag gtcacgtgaa gagagaaatt
cggccacagt 720tgtggtccta aacaaggact gaaaaagatc gcagaaacta
gcaacgtcgg atagcggtat 780ggttgaacca aagtccagtt tttagtcata
gccggttgag ggatgatctg ccgttcagtg 840ttctgcgttc aataatacgc
atatatatat atatcaaggt atcaacacaa tacttgaaca 900ggttaggctc
gtactaaagc ctggaagttt ttatctttct gtaacatttt tctttcttga
960ggtgtgtgtg tgtatagatt agcagggaat tatctaagat atg
1003351003DNASaccharomyces cerevisiae 35cgactaacct ttttcttggc
agtatggaga ctgactaggt ctcccaaaca ttcattgtaa 60ctgctgttta aagattttgt
tctaacctaa attcaagtga gaagctgaac atgtgtctct 120acttatgata
tcacgacagc aaatactaat cttgccataa atagtctagc gttttgcaac
180ttacctctag atatatttta tttcttgagg aaccgttttc gtcggtaata
acaaaatact 240actgaaacgc cacagcattg agagaatacg ttatcgatta
cggctttctt ctcgctccag 300atgtcgcggg taagatattc acctcaaact
tttcttgttg agtgtcgtca caaatctaga 360acctacatgc catctcaacg
atttttctgg agaaaggcct cactccgttc cgtacgtaat 420gcatagataa
agtatcagga tcttcacgat gctcgagagt tacttagtag tctgagttta
480tgcgaaaaaa actccgccgt tgtaataatc gggaatacac agaagtagta
ctgcactatc 540actgggatac tcaaaaacct tctttttaac ttttctatcc
cacaaataga acataggaaa 600gaacattgac tcctccactt gaagttaaat
tacaggaaca aacgcctaac tataatttcg 660acattgttgc atcaacgaat
cgaccgaaag aaaaatctgg agttgcagtt atcacttgta 720tgtgcactaa
gatttatatt tttactcctg agatctgcca aatcggtagc ttattgaact
780gcgttccttt ttcccctgag ttctcgaggt acctgcggct ttgtctgtgc
catctccccc 840actttaaagt accccacgtt actaccgcgt ttttccccac
ccccggctta ataaattagc 900tatatcttgt tgacttaaat acggagaaaa
gaagaaaacc ttcaagaaat gcttcattgt 960cttgtcaaaa gagctaaagt
aaaagagctc tagttcgaag atg 1003361003DNASaccharomyces cerevisiae
36ctgtcgagtg ctggtggcgc ttgatagcgg tattgtactt aataggtatc gccttcattg
60gtttgaaatt ctcgatcaaa ttctgcagga tagggtcctg agcaggcaca gaggcttgtt
120tcagagtaag ggtctccaat ttctgtttaa gggcatgatt ttccgattct
ttcgcttgaa 180gttcattttg gaggcggttt agtttcgttt cgtagttttt
aacgacctta ctcagtgact 240ctatggtttc ctccagcact tgcaacttgt
tgtttttgcg ttctctgtag gccctttggg 300cgtctctatt ctgccgtttt
ttcttctgta gctcctcatc attttcttca gtttcgtaat 360cttcgtgaag
tcgatgaact cttttcctcc gctgagctgc tctatggggc aaccttggag
420gcagtttcca atttttggac acgtggatct tggacaaaag agcgaggtgg
ctttcctcag 480actccttagg ttttatcaga ggtagagcca tgactgtgat
aatatgctag ttacactgtt 540tatgttgtgt gaacttgttg taatatggtt
aacttcactt tcagtgattg atatgatagc 600gacatcactg ccgtgcaaaa
agaccattcc attactgcac ctttttgtcc ttttccgtgg 660aataaaagtt
cactcgtcag ttccatgcat tctggaaaaa aatgatctga aagatgccac
720agttgtgggg cccgcccggc ccaataggta aactaaaata caatagaagg
ggtactgagt 780gcacgtgact tatttttttt ttttggtttt aggtttcgct
tttttcacct ttttctactt 840tctaacacca cagttttggg cgggaagcgg
aaacgccata gttgtaggtc actggcgtga 900gtcaaggccg ggcagccaat
gactaagaac acgaggtaac ttgaatttaa ctatttataa 960ccagtggtag
ttacgaagac aaattgtttt gttcgtcaat atg 1003371003DNASaccharomyces
cerevisiae 37caattttgaa aattgacata taaaactctg attctgcaaa gcctcttgcc
ccaaaaagat 60tcaatagcat gcttaaaacg aacgcgattg caacccaggc agcaggattg
accgatggtg 120cccaataaga gatggtaagc gaacatccta ccagctcgct
agggtaagat atgagccaca 180tcaataagta gttaattcct atactgaagc
cgactgatgg gtctataaat ctactggcgt 240gcagcgcata ggagccggaa
acagggtact gacatgaaag ttcggcccca cactgaacaa 300cacagagcat
agatgcgccg accaacaaga aaccaattag caaagaagca ggtccggacg
360ctagtgattc ccctaaacca atgaaaagac ccgtccctag agtacctcca
atggctatca 420tggtcaggtg tctttgatct aggacctttt tatacggctg
gttggctaag tcccattgta 480ttttttcttc ctctgaaagc tccgaataca
gcaaaccttc ttggatcgct ggtgaatcat 540ttcgcttgaa agagttcttg
aatctgtgta ttagattatt tccttcattc gaaggcgtac 600ttaacacgct
agtggaaaac tgaaccttcg aaagtttggt agattcaaat tcatccattt
660cgggtatctt tcatggtatc ctgatggaga actcttgacg gtgattactt
aaccacccaa 720atctgtaaaa tatatacgca tttatttcct ggcatctctt
cgctatcccc gcaaaaagtt 780aatgaaaaaa agccattcct gcaggaactg
tggaaaagta aaaactgtgg catgaaagta 840ctaccaaatt gtggcgaggt
attcaatggt aatagcaaaa cagcaacaac gccaatttga 900aaatgtacta
ctaaggaaga taatttaaac agacaagttg tagaagaaat ctgaatcagc
960aacttttagt gtgcaacttg aaaaagtaat caaaaaacag atg
1003381003DNASaccharomyces cerevisiae 38cagttttctt tttgaatgaa
ggagaaaagt gttctagaaa tatgttttag ttctaagtat 60cgttgtctgg agcaagcaaa
gaaaagcaga cctgtatgtg caactttctt gttttccatt 120aaaatctcaa
gaaccggccc cttgactgtg ctcatatatc aagtatcggt tttaaagtcc
180cctaattgtg ataattgcaa gaatgacagt caaaaagata tttttctttt
ctttcttttt 240acctatcctt catgaaaata cctatgctga tactgaatat
cttcatttgc agctatgcaa 300tatttcagat actgcatttt acccaagagt
ttctttactg gtggttggat tttttttaag 360acgagaagat taccagtcct
ggatgttcat gaatgtgcct attgcgtcat cagcgactcc 420gcgccaagaa
aagaaatact cgcatgctcg gatttataga atcgcataat ggaaatagct
480tccttaaagg atggagataa ggaaaaaatg acaagaacaa ggaagtactt
gggagctttc 540tccgacatgt taaatatttg aaactcacgc agctctcctg
gaaagttgca tctggaaagg 600taaggttgtt ttttgttcag acatcaaaat
cggctcttgt agacaatgtc acagatccac 660aagttgaaaa aaaagtttcg
atgaactgga taaggggaga ggtcacaaac aaagtgtagg 720gggtgagtag
tatagtggta acttgttcat ttacctaaat aacattctca ctagaaaaga
780gatattgaaa gactcctcgt ttacctaact tggctggttc ttaggtatat
aatagaaaca 840tggatctggc aggttattac ttacagtttg cattcccacc
aggtaacaat tttcattata 900tatagttatt ttttaccacc acttcttctt
ctttcacttt gtcttgcaat agaaatacca 960aaacaaggtg aggaaaaaat
cgaatctcaa cgataaaaat atg 1003391003DNASaccharomyces cerevisiae
39cggttggatc ctcacttgtt ttaacacctc caaataacaa gtttttaatg aaggacattt
60gttctctata atattccgat gtacgtgtgt gtggctgatg agatttagac tggttagact
120atttgacgcg tctattatag cttactgcaa caagaaaatg atcgttgata
tataaactct 180cagatgtata tatcgttctg gaaacatcga gcataataca
atacaattca acaaaaatgc 240gagaaggcac tgatgtcttg tcgttaaaga
accaaaaacg cggacactac gaccgtctta 300tttccggtag aaaaagggta
catacagttg aaggaacgaa gaaaattaaa attagaaaaa 360aaagtaaaat
aaaacaagga aggtagggta atatggtctc gtttcctttg tcgctccgca
420aataaaggag cttattcccg cacgctcaca tggtaatttg cgccaaatca
cggatgtgga 480aaactgatca cgtgcttcga tcgccaacta ctgagcgtcg
tcccacactg atctggcaca 540gcttacctcg ccttgaaaat tttaatctgt
cctgctcgtt tgttgtatat tgcttcttct 600cagaatatgc ccgcgataac
tgacaaagag ggttcgacgt ttcagagatt ctactcttga 660ccactgtttc
gtgtagccgc tcaaggttta tttctttctt ctttaatgtt cttggcactt
720aggcggctcc gtcctccgtc tgaaattgcc gatcctatta tttgcggagg
gctccttaga 780agggctcctt agtaagcagt ttgcgttcct gatataactc
cgttcagaac aaggataaag 840tcgcaataac cattactaag cacagtgttg
taagtaggac aactcgaacc tatataaggg 900ttgtgaactg tgcttgattc
ttgcccatca tatgcaaaaa agtacgtact tgatatatac 960aacaactgta
gttcagtata gcgaagttta aatttagaag atg 1003401003DNASaccharomyces
cerevisiae 40tggaaatgta aaggataatg agtgagcata taaaatggaa gaaaaaataa
taataggatt 60atgtataaaa tatcgattcc cttttgtaga tttcgagatc ttcgaggaga
acttctagca 120cgttgtttac atatctaatg ttgtaacacg gcctttgtta
tcttgtgaat atatccatca 180cctacgtcgc ttgaggtttc gtatcaaaga
aagagagttc aaacataatc atacgtgttt 240cactccaaat ttgatcacaa
atggccaccc attatagctt gttattattt acttcttctt 300gagttcattt
tattgaaaga tgcccgctgg gaacaaaaca taaataaaat tcaagagtac
360aatatagcgt agcatagttg cgagcaaggg aaggcaacac agttaaaatg
acaatgttag 420aaggaaaaac taaatcaagc tcccatagga aatttcttgt
gtcagtaata tgtctcgaga 480aaataataat aattgaatct atttattgaa
aagtaaatat ctcgtaaccc ggatgctttg 540ggcggtcggg ttttgctact
cgtcatccga tgagaaaaac tgttcccttt tgccccaggt 600ttccattcat
ccgagcgatc acttatctga cttcgtcact ttttcatttc atccgaaaca
660atcaaaactg aagccaatca ccacaaaatt aacactcaac gtcatctttc
actacccttt 720acagaagaaa atatccatag tccggactag catcccagta
tgtgactcaa tattggtgca 780aaagagaaaa gcataagtca gtccaaagtc
cgcccttaac caggcacatc ggaattcaca 840aaacgtttct ttattatata
aaggagctgc ttcactggca aaattcttat tatttgtctt 900ggcttgctaa
tttcatctta tccttttttt cttttcacac ccaaatacct aacaattgag
960agaaaactct tagcataaca taacaaaaag tcaacgaaaa atg
1003411003DNASaccharomyces cerevisiae 41ttcctaagcc tttgagatat
atcaaaattg ctgcctttac caacgacgaa agtaaattgg 60aagagcttcg tgagaaatac
ccaaatcatt tgattggcgg agcagatctt gttgccaaaa 120ttaaaagcgg
tgaaattagc gttgattttg acaaagcatt tgcaactcct gatattgttc
180cagctttaca atcgcaggtg gccaggatac ttggtcctcg tggggtttta
ccctctgtca 240aaaaaggtac ggtgagtgat gatataagct ccttacttca
agaaagtcta ggatctatgc 300ctttcaggca aaggggaaac tctatcagca
ttggagtcgg caaatgctac ttcactgatc 360gagaaatttt acaaaacatt
atatccgcta gagcggcatt taaaactgct gttgataatc 420aaaaatctaa
gaaaccaaat atactgagta aaaccacatt atcaagcaca catggccccg
480gtattgtaat cgattttgct taatcagcga atcagttttg tcttgttgga
cttttttaat 540tgtatatata tatatactgg tgcattttct ttctttttat
ataaaatcat gtacatacaa 600agtaaccgtc tttttaaagt ggaataaagt
aactgctctt ttatttttga ataatagtga 660aagtgtatgt tgcataatca
ttagatacct atgtacaaga agtattcaaa tttatctgaa 720tagtatgtga
aatttcaatg agacaaaaac cacctacctt tgagcttcct ccttttcaat
780atcttagagc cctgtttgct cttggcccta gccaagaaac caaaagttct
cttccgtttc 840aatgtactag gttgataggt gttacccctt gatttccatc
ttctttgacc tattaagcca 900aatcccaacg gtgtgaagga cggagtcttc
aatggtgaac tgtttaagag cataccggtt 960tgtgggcgta acacagatag
agcagagaaa gaagatatag atg 1003421716DNASaccharomyces cerevisiae
42atggattcta gaacagttgg tatattagga gggggacaat tgggacgtat gattgttgag
60gcagcaaaca ggctcaacat taagacggta atactagatg ctgaaaattc tcctgccaaa
120caaataagca actccaatga ccacgttaat ggctcctttt ccaatcctct
tgatatcgaa 180aaactagctg aaaaatgtga tgtgctaacg attgagattg
agcatgttga tgttcctaca 240ctaaagaatc ttcaagtaaa acatcccaaa
ttaaaaattt acccttctcc agaaacaatc 300agattgatac aagacaaata
tattcaaaaa gagcatttaa tcaaaaatgg tatagcagtt 360acccaaagtg
ttcctgtgga acaagccagt gagacgtccc tattgaatgt tggaagagat
420ttgggttttc cattcgtctt gaagtcgagg actttggcat acgatggaag
aggtaacttc 480gttgtaaaga ataaggaaat gattccggaa gctttggaag
tactgaagga tcgtcctttg 540tacgccgaaa aatgggcacc atttactaaa
gaattagcag tcatgattgt gagatctgtt 600aacggtttag tgttttctta
cccaattgta gagactatcc acaaggacaa tatttgtgac 660ttatgttatg
cgcctgctag agttccggac tccgttcaac ttaaggcgaa gttgttggca
720gaaaatgcaa tcaaatcttt tcccggttgt ggtatatttg gtgtggaaat
gttctattta 780gaaacagggg aattgcttat taacgaaatt gccccaaggc
ctcacaactc tggacattat 840accattgatg cttgcgtcac ttctcaattt
gaagctcatt tgagatcaat attggatttg 900ccaatgccaa agaatttcac
atctttctcc accattacaa cgaacgccat tatgctaaat 960gttcttggag
acaaacatac aaaagataaa gagctagaaa cttgcgaaag agcattggcg
1020actccaggtt cctcagtgta cttatatgga aaagagtcta gacctaacag
aaaagtaggt 1080cacataaata ttattgcctc cagtatggcg gaatgtgaac
aaaggctgaa ctacattaca 1140ggtagaactg atattccaat caaaatctct
gtcgctcaaa agttggactt ggaagcaatg 1200gtcaaaccat tggttggaat
catcatggga tcagactctg acttgccggt aatgtctgcc 1260gcatgtgcgg
ttttaaaaga ttttggcgtt ccatttgaag tgacaatagt ctctgctcat
1320agaactccac ataggatgtc agcatatgct atttccgcaa gcaagcgtgg
aattaaaaca 1380attatcgctg gagctggtgg ggctgctcac ttgccaggta
tggtggctgc aatgacacca 1440cttcctgtca tcggtgtgcc cgtaaaaggt
tcttgtctag atggagtaga ttctttacat 1500tcaattgtgc aaatgcctag
aggtgttcca gtagctaccg tcgctattaa taatagtacg 1560aacgctgcgc
tgttggctgt cagactgctt ggcgcttatg attcaagtta tacaacgaaa
1620atggaacagt ttttattaaa gcaagaagaa gaagttcttg tcaaagcaca
aaagttagaa 1680actgtcggtt acgaagctta tctagaaaac aagtaa
1716434179DNASaccharomyces cerevisiae 43atgactaacg aaaaggtctg
gatagagaag ttggataatc caactctttc agtgttacca 60catgactttt tacgcccaca
acaagaacct tatacgaaac aagctacata ttcgttacag 120ctacctcagc
tcgatgtgcc tcatgatagt ttttctaaca aatacgctgt cgctttgagt
180gtatgggctg cattgatata tagagtaacc ggtgacgatg atattgttct
ttatattgcg 240aataacaaaa tcttaagatt caatattcaa ccaacgtggt
catttaatga gctgtattct 300acaattaaca atgagttgaa caagctcaat
tctattgagg ccaatttttc ctttgacgag 360ctagctgaaa aaattcaaag
ttgccaagat ctggaaagga cccctcagtt gttccgtttg 420gcctttttgg
aaaaccaaga tttcaaatta gacgagttca agcatcattt agtggacttt
480gctttgaatt tggataccag taataatgcg catgttttga acttaattta
taacagctta 540ctgtattcga atgaaagagt aaccattgtt gcggaccaat
ttactcaata tttgactgct 600gcgctaagcg atccatccaa ttgcataact
aaaatctctc tgatcaccgc atcatccaag 660gatagtttac ctgatccaac
taagaacttg ggctggtgcg atttcgtggg gtgtattcac 720gacattttcc
aggacaatgc tgaagccttc ccagagagaa cctgtgttgt ggagactcca
780acactaaatt ccgacaagtc ccgttctttc acttatcgcg acatcaaccg
cacttctaac 840atagttgccc attatttgat taaaacaggt atcaaaagag
gtgatgtagt gatgatctat 900tcttctaggg gtgtggattt gatggtatgt
gtgatgggtg tcttgaaagc cggcgcaacc 960ttttcagtta tcgaccctgc
atatccccca gccagacaaa ccatttactt aggtgttgct 1020aaaccacgtg
ggttgattgt tattagagct gctggacaat tggatcaact agtagaagat
1080tacatcaatg atgaattgga gattgtttca agaatcaatt ccatcgctat
tcaagaaaat 1140ggtaccattg aaggtggcaa attggacaat ggcgaggatg
ttttggctcc atatgatcac 1200tacaaagaca ccagaacagg tgttgtagtt
ggaccagatt ccaacccaac cctatctttc 1260acatctggtt ccgaaggtat
tcctaagggt gttcttggta gacatttttc cttggcttat 1320tatttcaatt
ggatgtccaa aaggttcaac ttaacagaaa atgataaatt cacaatgctg
1380agcggtattg cacatgatcc aattcaaaga gatatgttta caccattatt
tttaggtgcc 1440caattgtatg tccctactca agatgatatt ggtacaccgg
gccgtttagc ggaatggatg 1500agtaagtatg gttgcacagt tacccattta
acacctgcca tgggtcaatt acttactgcc 1560caagctacta caccattccc
taagttacat catgcgttct ttgtgggtga cattttaaca 1620aaacgtgatt
gtctgaggtt acaaaccttg gcagaaaatt gccgtattgt taatatgtac
1680ggtaccactg aaacacagcg tgcagtttct tatttcgaag ttaaatcaaa
aaatgacgat 1740ccaaactttt tgaaaaaatt gaaagatgtc atgcctgctg
gtaaaggtat gttgaacgtt 1800cagctactag ttgttaacag gaacgatcgt
actcaaatat gtggtattgg cgaaataggt 1860gagatttatg ttcgtgcagg
tggtttggcc gaaggttata gaggattacc agaattgaat 1920aaagaaaaat
ttgtgaacaa ctggtttgtt gaaaaagatc actggaatta tttggataag
1980gataatggtg aaccttggag acaattctgg ttaggtccaa gagatagatt
gtacagaacg 2040ggtgatttag gtcgttatct accaaacggt gactgtgaat
gttgcggtag ggctgatgat 2100caagttaaaa ttcgtgggtt cagaatcgaa
ttaggagaaa tagatacgca catttcccaa 2160catccattgg taagagaaaa
cattacttta gttcgcaaaa atgccgacaa tgagccaaca 2220ttgatcacat
ttatggtccc aagatttgac aagccagatg acttgtctaa gttccaaagt
2280gatgttccaa aggaggttga aactgaccct atagttaagg gcttaatcgg
ttaccatctt 2340ttatccaagg acatcaggac tttcttaaag aaaagattgg
ctagctatgc tatgccttcc 2400ttgattgtgg ttatggataa actaccattg
aatccaaatg gtaaagttga taagcctaaa 2460cttcaattcc caactcccaa
gcaattaaat ttggtagctg aaaatacagt ttctgaaact 2520gacgactctc
agtttaccaa tgttgagcgc gaggttagag acttatggtt aagtatatta
2580cctaccaagc cagcatctgt atcaccagat gattcgtttt tcgatttagg
tggtcattct 2640atcttggcta ccaaaatgat ttttacctta aagaaaaagc
tgcaagttga tttaccattg 2700ggcacaattt tcaagtatcc aacgataaag
gcctttgccg cggaaattga cagaattaaa 2760tcatcgggtg gatcatctca
aggtgaggtc gtcgaaaatg tcactgcaaa ttatgcggaa 2820gacgccaaga
aattggttga gacgctacca agttcgtacc cctctcgaga atattttgtt
2880gaacctaata gtgccgaagg aaaaacaaca attaatgtgt ttgttaccgg
tgtcacagga 2940tttctgggct cctacatcct tgcagatttg ttaggacgtt
ctccaaagaa ctacagtttc 3000aaagtgtttg cccacgtcag ggccaaggat
gaagaagctg catttgcaag attacaaaag 3060gcaggtatca cctatggtac
ttggaacgaa aaatttgcct caaatattaa agttgtatta 3120ggcgatttat
ctaaaagcca atttggtctt tcagatgaga agtggatgga tttggcaaac
3180acagttgata taattatcca taatggtgcg ttagttcact gggtttatcc
atatgccaaa 3240ttgagggatc caaatgttat ttcaactatc aatgttatga
gcttagccgc cgtcggcaag 3300ccaaagttct ttgactttgt ttcctccact
tctactcttg acactgaata ctactttaat 3360ttgtcagata aacttgttag
cgaagggaag ccaggcattt tagaatcaga cgatttaatg 3420aactctgcaa
gcgggctcac tggtggatat ggtcagtcca aatgggctgc tgagtacatc
3480attagacgtg caggtgaaag gggcctacgt gggtgtattg tcagaccagg
ttacgtaaca 3540ggtgcctctg ccaatggttc ttcaaacaca gatgatttct
tattgagatt tttgaaaggt 3600tcagtccaat taggtaagat tccagatatc
gaaaattccg tgaatatggt tccagtagat 3660catgttgctc gtgttgttgt
tgctacgtct ttgaatcctc ccaaagaaaa tgaattggcc 3720gttgctcaag
taacgggtca cccaagaata ttattcaaag actacttgta tactttacac
3780gattatggtt acgatgtcga aatcgaaagc tattctaaat ggaagaaatc
attggaggcg 3840tctgttattg acaggaatga agaaaatgcg ttgtatcctt
tgctacacat ggtcttagac 3900aacttacctg aaagtaccaa agctccggaa
ctagacgata ggaacgccgt ggcatcttta 3960aagaaagaca ccgcatggac
aggtgttgat tggtctaatg gaataggtgt tactccagaa 4020gaggttggta
tatatattgc atttttaaac aaggttggat ttttacctcc accaactcat
4080aatgacaaac ttccactgcc aagtatagaa ctaactcaag cgcaaataag
tctagttgct 4140tcaggtgctg gtgctcgtgg aagctccgca gcagcttaa
417944675DNASaccharomyces cerevisiae 44atgtctgtta ttaatttcac
aggtagttct ggtccattgg tgaaagtttg cggcttgcag 60agcacagagg ccgcagaatg
tgctctagat tccgatgctg acttgctggg tattatatgt 120gtgcccaata
gaaagagaac aattgacccg gttattgcaa ggaaaatttc aagtcttgta
180aaagcatata aaaatagttc aggcactccg aaatacttgg ttggcgtgtt
tcgtaatcaa 240cctaaggagg atgttttggc tctggtcaat gattacggca
ttgatatcgt ccaactgcat 300ggagatgagt cgtggcaaga ataccaagag
ttcctcggtt tgccagttat taaaagactc 360gtatttccaa aagactgcaa
catactactc agtgcagctt cacagaaacc tcattcgttt 420attcccttgt
ttgattcaga agcaggtggg acaggtgaac ttttggattg gaactcgatt
480tctgactggg ttggaaggca agagagcccc gaaagcttac attttatgtt
agctggtgga 540ctgacgccag aaaatgttgg tgatgcgctt agattaaatg
gcgttattgg tgttgatgta 600agcggaggtg tggagacaaa tggtgtaaaa
gactctaaca aaatagcaaa tttcgtcaaa 660aatgctaaga aatag
675451095DNASaccharomyces cerevisiae 45atgtctgccc ctaagaagat
cgtcgttttg ccaggtgacc acgttggtca agaaatcaca 60gccgaagcca ttaaggttct
taaagctatt tctgatgttc gttccaatgt caagttcgat 120ttcgaaaatc
atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat
180gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt
gggtggtcct 240aaatggggta ccggtagtgt tagacctgaa caaggtttac
taaaaatccg taaagaactt 300caattgtacg ccaacttaag accatgtaac
tttgcatccg actctctttt agacttatct 360ccaatcaagc cacaatttgc
taaaggtact gacttcgttg ttgtcagaga attagtggga 420ggtatttact
ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa
480caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat
ggccctacaa 540catgagccac cattgcctat ttggtccttg gataaagcta
atgttttggc ctcttcaaga 600ttatggagaa aaactgtgga ggaaaccatc
aagaacgaat tccctacatt gaaggttcaa 660catcaattga ttgattctgc
cgccatgatc ctagttaaga acccaaccca cctaaatggt 720attataatca
ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca
780ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa
gaacaccgca 840tttggtttgt acgaaccatg ccacggttct gctccagatt
tgccaaagaa taaggtcaac 900cctatcgcca ctatcttgtc tgctgcaatg
atgttgaaat tgtcattgaa cttgcctgaa 960gaaggtaagg ccattgaaga
tgcagttaaa aaggttttgg atgcaggtat cagaactggt 1020gatttaggtg
gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag
1080aaaatccttg cttaa 109546804DNASaccharomyces cerevisiae
46atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
60ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
120accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt
actaaaaaca 180catgtggata tcttgactga tttttccatg gagggcacag
ttaagccgct aaaggcatta 240tccgccaagt acaatttttt actcttcgaa
gacagaaaat ttgctgacat tggtaataca 300gtcaaattgc agtactctgc
gggtgtatac agaatagcag aatgggcaga cattacgaat 360gcacacggtg
tggtgggccc aggtattgtt agcggtttga agcaggcggc ggaagaagta
420acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg
ctccctagct 480actggagaat atactaaggg tactgttgac attgcgaaga
gcgacaaaga ttttgttatc 540ggctttattg ctcaaagaga catgggtgga
agagatgaag gttacgattg gttgattatg 600acacccggtg tgggtttaga
tgacaaggga gacgcattgg gtcaacagta tagaaccgtg 660gatgatgtgg
tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag
720ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga
agcatatttg 780agaagatgcg gccagcaaaa ctaa 80447663DNASaccharomyces
cerevisiae 47atgacagagc agaaagccct agtaaagcgt attacaaatg aaaccaagat
tcagattgcg 60atctctttaa agggtggtcc cctagcgata gagcactcga tcttcccaga
aaaagaggca 120gaagcagtag cagaacaggc cacacaatcg caagtgatta
acgtccacac aggtataggg 180tttctggacc atatgataca tgctctggcc
aagcattccg gctggtcgct aatcgttgag 240tgcattggtg acttacacat
agacgaccat cacaccactg aagactgcgg gattgctctc 300ggtcaagctt
ttaaagaggc cctaggggcc gtgcgtggag taaaaaggtt tggatcagga
360tttgcgcctt tggatgaggc actttccaga gcggtggtag atctttcgaa
caggccgtac 420gcagttgtcg aacttggttt gcaaagggag aaagtaggag
atctctcttg cgagatgatc 480ccgcattttc ttgaaagctt tgcagaggct
agcagaatta ccctccacgt tgattgtctg 540cgaggcaaga atgatcatca
ccgtagtgag agtgcgttca aggctcttgc ggttgccata 600agagaagcca
cctcgcccaa tggtaccaac gatgttccct ccaccaaagg tgttcttatg 660tag
66348736DNAAequorea victoria 48aagctttatt aaaatgtcta aaggtgaaga
attattcact ggtgttgtcc caattttggt 60tgaattagat ggtgatgtta atggtcacaa
attttctgtc tccggtgaag gtgaaggtga 120tgctacttac ggtaaattga
ccttaaaatt tatttgtact actggtaaat tgccagttcc 180atggccaacc
ttagtcacta ctttcggtta tggtgttcaa tgttttgcta gatacccaga
240tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg
ttcaagaaag 300aactattttt ttcaaagatg acggtaacta caagaccaga
gctgaagtca agtttgaagg 360tgatacctta gttaatagaa tcgaattaaa
aggtattgat tttaaagaag atggtaacat 420tttaggtcac aaattggaat
acaactataa ctctcacaat gtttacatca tggctgacaa 480acaaaagaat
ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt
540tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag
tcttgttacc 600agacaaccat tacttatcca ctcaatctgc cttatccaaa
gatccaaacg aaaagagaga 660ccacatggtc ttgttagaat ttgttactgc
tgctggtatt acccatggta tggatgaatt 720gtacaaataa ctgcag
73649945DNARenilla reniformis 49atggcttcga aagtttatga tccagaacaa
aggaaacgga tgataactgg tccgcagtgg 60tgggccagat gtaaacaaat gaatgttctt
gattcattta ttaattatta tgattcagaa 120aaacatgcag aaaatgctgt
tattttttta catggtaacg cggcctcttc ttatttatgg 180cgacatgttg
tgccacatat tgagccagta gcgcggtgta ttataccaga ccttattggt
240atgggcaaat caggcaaatc tggtaatggt tcttataggt tacttgatca
ttacaaatat 300cttactgcat ggtttgaact tcttaattta ccaaagaaga
tcatttttgt cggccatgat 360tggggtgctt gtttggcatt tcattatagc
tatgagcatc aagataagat caaagcaata 420gttcacgctg aaagtgtagt
agatgtgatt gaatcatggg atgaatggcc tgatattgaa 480gaagatattg
cgttgatcaa atctgaagaa ggagaaaaaa tggttttgga gaataacttc
540ttcgtggaaa ccatgttgcc atcaaaaatc atgagaaagt tagaaccaga
agaatttgca 600gcatatcttg aaccattcaa agagaaaggt gaagttcgtc
gtccaacatt atcatggcct 660cgtgaaatcc cgttagtaaa aggtggtaaa
cctgacgttg tacaaattgt taggaattat 720aatgcttatc tacgtgcaag
tgatgattta ccaaaaatgt ttattgaatc ggacccagga 780ttcttttcca
atgctattgt tgaaggtgcc aagaagtttc ctaatactga atttgtcaaa
840gtaaaaggtc ttcatttttc gcaagaagat gcacctgatg aaatgggaaa
atatatcaaa 900tcgttcgttg agcgagttct caaaaatgaa caaagatcta tctag
945502387DNAPhotinus pyralis 50ctgcagaaat aactaggtac taagcccgtt
tgtgaaaagt ggccaaaccc ataaatttgg 60caattacaat aaagaagcta aaattgtggt
caaactcaca aacattttta ttatatacat 120tttagtagct gatgcttata
aaagcaatat ttaaatcgta aacaacaaat aaaataaaat 180ttaaacgatg
tgattaagag ccaaaggtcc tctagaaaaa ggtatttaag caacggaatt
240cctttgtgtt acattcttga atgtcgctcg cagtgacatt agcattccgg
tactgttggt 300aaaatggaag acgccaaaaa cataaagaaa ggcccggcgc
cattctatcc tctagaggat 360ggaaccgctg gagagcaact gcataaggct
atgaagagat acgccctggt tcctggaaca 420attgcttttg tgagtatttc
tgtctgattt ctttcgagtt aacgaaatgt tcttatgttt 480ctttagacag
atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc
540gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
aatcgtcgta 600tgcagtgaaa actctcttca attctttatg ccggtgttgg
gcgcgttatt tatcggagtt 660gcagttgcgc ccgcgaacga catttataat
gaacgtaagc accctcgcca tcagaccaaa 720gggaatgacg tatttaattt
ttaaggtgaa ttgctcaaca gtatgaacat ttcgcagcct 780accgtagtgt
ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaaatta
840ccaataatcc agaaaattat tatcatggat tctaaaacgg attaccaggg
atttcagtcg 900atgtacacgt tcgtcacatc tcatctacct cccggtttta
atgaatacga ttttgtacca 960gagtcctttg atcgtgacaa aacaattgca
ctgataatga attcctctgg atctactggg 1020ttacctaagg gtgtggccct
tccgcataga actgcctgcg tcagattctc gcatgccagg 1080tatgtcgtat
aacaagagat taagtaatgt tgctacacac attgtagaga tcctattttt
1140ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
tcacggtttt 1200ggaatgttta ctacactcgg atatttgata tgtggatttc
gagtcgtctt aatgtataga 1260tttgaagaag agctgttttt acgatccctt
caggattaca aaattcaaag tgcgttgcta 1320gtaccaaccc tattttcatt
cttcgccaaa agcactctga ttgacaaata cgatttatct 1380aatttacacg
aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt
1440gcaaaacggt gagttaagcg cattgctagt atttcaaggc tctaaaacgg
cgcgtagctt 1500ccatcttcca gggatacgac aaggatatgg gctcactgag
actacatcag ctattctgat 1560tacacccgag ggggatgata aaccgggcgc
ggtcggtaaa gttgttccat tttttgaagc 1620gaaggttgtg gatctggata
ccgggaaaac gctgggcgtt aatcagagag gcgaattatg 1680tgtcagagga
cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
1740gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
aagacgaaca 1800cttcttcata gttgaccgct tgaagtcttt aattaaatac
aaaggatatc aggtaatgaa 1860gatttttaca tgcacacacg ctacaatacc
tgtaggtggc ccccgctgaa ttggaatcga 1920tattgttaca acaccccaac
atcttcgacg cgggcgtggc aggtcttccc gacgatgacg 1980ccggtgaact
tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
2040agatcgtgga ttacgtcgcc agtaaatgaa ttcgttttac gttactcgta
ctacaattct 2100tttcataggt caagtaacaa ccgcgaaaaa gttgcgcgga
ggagttgtgt ttgtggacga 2160agtaccgaaa ggtcttaccg gaaaactcga
cgcaagaaaa atcagagaga tcctcataaa 2220ggccaagaag ggcggaaagt
ccaaattgta aaatgtaact gtattcagcg atgacgaaat 2280tcttagctat
tgtaatatta tatgcaaatt gatgaatggt aattttgtaa ttgtgggtca
2340ctgtactatt ttaacgaata ataaaatcag gtataggtaa ctaaaaa
2387513075DNAEscherichia coli 51atgaccatga ttacggattc actggccgtc
gttttacaac gtcgtgactg ggaaaaccct 60ggcgttaccc aacttaatcg ccttgcagca
catccccctt tcgccagctg gcgtaatagc 120gaagaggccc gcaccgatcg
cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc 180tttgcctggt
ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
240gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
tgcgcccatc 300tacaccaacg tgacctatcc cattacggtc aatccgccgt
ttgttcccac ggagaatccg 360acgggttgtt actcgctcac atttaatgtt
gatgaaagct ggctacagga aggccagacg 420cgaattattt ttgatggcgt
taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc 480ggttacggcc
aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc
540ggagaaaacc gcctcgcggt gatggtgctg cgctggagtg acggcagtta
tctggaagat 600caggatatgt ggcggatgag cggcattttc cgtgacgtct
cgttgctgca taaaccgact 660acacaaatca gcgatttcca tgttgccact
cgctttaatg atgatttcag ccgcgctgta 720ctggaggctg aagttcagat
gtgcggcgag ttgcgtgact acctacgggt aacagtttct 780ttatggcagg
gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc
840gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga
aaacccgaaa 900ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg
ttgaactgca caccgccgac 960ggcacgctga ttgaagcaga agcctgcgat
gtcggtttcc gcgaggtgcg gattgaaaat 1020ggtctgctgc tgctgaacgg
caagccgttg ctgattcgag gcgttaaccg tcacgagcat 1080catcctctgc
atggtcaggt catggatgag cagacgatgg tgcaggatat cctgctgatg
1140aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc
gctgtggtac 1200acgctgtgcg accgctacgg cctgtatgtg gtggatgaag
ccaatattga aacccacggc 1260atggtgccaa tgaatcgtct gaccgatgat
ccgcgctggc taccggcgat gagcgaacgc 1320gtaacgcgaa tggtgcagcg
cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg 1380aatgaatcag
gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat
1440ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac
cgatattatt 1500tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc
cggctgtgcc gaaatggtcc 1560atcaaaaaat ggctttcgct acctggagag
acgcgcccgc tgatcctttg cgaatacgcc 1620cacgcgatgg gtaacagtct
tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat 1680ccccgtttac
agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat
1740gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc
gaacgatcgc 1800cagttctgta tgaacggtct ggtctttgcc gaccgcacgc
cgcatccagc gctgacggaa 1860gcaaaacacc agcagcagtt tttccagttc
cgtttatccg ggcaaaccat cgaagtgacc 1920agcgaatacc tgttccgtca
tagcgataac gagctcctgc actggatggt ggcgctggat 1980ggtaagccgc
tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg taaacagttg
2040attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct
cacagtacgc 2100gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc
acatcagcgc ctggcagcag 2160tggcgtctgg cggaaaacct cagtgtgacg
ctccccgccg cgtcccacgc catcccgcat 2220ctgaccacca gcgaaatgga
tttttgcatc gagctgggta ataagcgttg gcaatttaac 2280cgccagtcag
gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg
2340ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag
tgaagcgacc 2400cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg
cgggccatta ccaggccgaa 2460gcagcgttgt tgcagtgcac ggcagataca
cttgctgatg cggtgctgat tacgaccgct 2520cacgcgtggc agcatcaggg
gaaaacctta tttatcagcc ggaaaaccta ccggattgat 2580ggtagtggtc
aaatggcgat taccgttgat gttgaagtgg cgagcgatac accgcatccg
2640gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa
ctggctcgga 2700ttagggccgc aagaaaacta tcccgaccgc cttactgccg
cctgttttga ccgctgggat 2760ctgccattgt cagacatgta taccccgtac
gtcttcccga gcgaaaacgg tctgcgctgc 2820gggacgcgcg aattgaatta
tggcccacac cagtggcgcg gcgacttcca gttcaacatc 2880agccgctaca
gtcaacagca actgatggaa accagccatc gccatctgct gcacgcggaa
2940gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga
cgactcctgg 3000agcccgtcag tatcggcgga attccagctg agcgccggtc
gctaccatta ccagttggtc 3060tggtgtcaaa aataa
3075521003DNASaccharomyces cerevisiae 52tctaaatatc gtcccgttac
cctcatcgct actgccgggc gttcccaacc ccttccgagg 60cagattatat ttatttatct
ttttaacaat ttgtccctat ataggcgggc cgaacacagc 120acgtgaaaga
actctcaggc ttccacaaca ggccatatca agtccacact tccctcatta
180ctagcatttt tacacaagca gtccacgcat tccgcaacag tgttcacctt
tcttttccct
240aacttatctt attatttgtt tgttgatcgc aaacagaccc ttttttttgg
tgagcgcatt 300ttacacagtc tcaagtgtcc tctactataa aaatacaacc
tcaaacagta cttactggcg 360tccgcagaat attactgcac tctcaatctg
taaagaatat aacaagtgac ccattccctt 420gttttattca acttcgaaat
ctctctggct atctttgcat gcttatctaa ctttcacgtg 480acccggatat
tgttctctgc tcaaaaaaga actgcaaaga aggaaataga aaaatagcat
540acaggctcaa agctatagtt ttctgaaaag ggacaccacg aatacagtag
tatcaaagag 600aaaaaagact tctttatact tacagaaggt tcattttctg
attctactct tccaatctag 660tgtcgcaaaa aaacaaaact tactggtatt
agctggcaaa aaaaaaaaga ataggttata 720attaaagaca tcttttctct
ataaattttt tcttagaaga aaaagccaaa aaaaaaaaac 780ctgcttcata
cccgccatca aatcttatag gccttataga ctgcatatta ccggattctt
840tctctccgtt aaccctacta ttctttcact atactttaac ctaaaataaa
aaaggaaaaa 900aaaaattaaa aacttccaaa aacttagaaa tttcaaatac
aaaacaaaaa gacacgtaaa 960gttagtgcaa tatttttttc ttacaatttt
ttgaaactcg atg 1003531003DNASaccharomyces cerevisiae 53agaacattat
gaagtaaaag gacaatcagc acgccttcca gacttttaag aaacattgat 60ggagccattg
atatcggcac cgtacctaac aacaacaaaa atgtctgctc ctgctacgct
120tgatgctgcc tgtatttttt gcaagattat taaaagtatg tcacattact
aataaagagc 180ttacactcac accaatgatg gcgatagtct ctatgtagta
catatacata aagcagaata 240ctaacaatcg atccgctatg caacaggcga
aattccatcc ttcaaattga ttgaaacaaa 300gtactcgtat gctttcttgg
acatccaacc tactgctgaa ggtcatgctt taatcattcc 360taagtaccat
ggtgcgaagt tgcatgacat cccggacgaa ttccttaccg atgctatgcc
420gattgccaag agactggcca aggcaatgaa gttggacact tataatgtgt
tgcagaataa 480tggtaaaatt gcgcatcaag aagtcgacca cgtccacttc
catttgattc ctaagagaga 540tgagaaaagt ggtttgattg tagggtggcc
agcccaagaa acggacttcg ataagttggg 600caagctacac aaggaattgc
ttgccaaact agaaggctcc gattagagta atgcatgcta 660tgatttcttt
tagcgttttc ttttcatatt taagtttata catatgggtg tgtttgcttc
720cattcatatc cactctataa tgctaacaat catgttacta gccatcgttg
ctatcgatac 780cttatgaaca gctctaactc cttccttttt aacctaccgc
cttttgcgca ctacccggca 840acccctcgaa tgtgaattcg gtggcaaaaa
aggacatata tcagggccgt gtatttaaga 900gatccgcagt tgatgtagtc
aattaatcta gttctattaa gatttgactc aacgccattg 960actcgataac
tggctcaact acaagacaga tatacaaatc atg 1003541003DNASaccharomyces
cerevisiae 54cgaactatag ggagattggt cctcaacaat atatttcttc cttgtcgacc
cattaggtgt 60atgggcgagt tgctgcaggg aagacaacga atccctagaa gcgcacttgt
aaagatcttg 120ctcttgtaag tagcgatgac ggtcatgaag atgaaagggt
gcggttgaag tcaagttgaa 180cgtgatcgga tactgatttc tagaaacagg
gggggctggt gttggttctg gcgaagccgc 240ttgctccgac tgctttgatg
tcacagcttt gggtttctcc gctacactgt ctaccttagg 300attattgatc
atgactcaag cttgcgatat gtgttggtgt catgcacaag acgccacaaa
360tgatagaaca gaaaagaaag tgaactaatc ttccaagacg aagaaaacca
aaatccggga 420tgagttgaaa gtcaaaaaga ctgtatatat aaatttcaac
ttttgtagaa gatgcagaaa 480aagaaaatga tatggtatgc agaaaaagaa
ataaaccgct attatcctcg cggtttgtca 540ttataacagg caattacact
agagaaagcc gcacacctcc ctccgtttct tttgccctgc 600gagtttttcc
ggaaaagaaa aaaaaaacga aaattaaaac ccgcctgccc cgcggggtgg
660gggaggggtc accggaacaa atcggataat ccctcgcctg ccttagatat
cgttctgaac 720ggctgaaatt atgaaagaag aagaacatca ctttacacgg
atcgcacgcc cataattctt 780tttttttttt ttttcatatc ttcgacgttt
gccactgcct tctctttttc tttctttttt 840ggcggccggt ggccaaacgc
gccaaaaccg aaacgcttat aaaatgtagt tgccttgctc 900tttcattcga
tatacatata agaacgctca ctgttatctt acattagaag ctagaaacta
960taacagtata acacagcaca agagaaccga gcagcccgcc atg
1003551003DNASaccharomyces cerevisiae 55agtaaaaaga atagtaatta
ttacaatgga gaagataaga taaagttagc ggcaacaaat 60aaagaactca tcagagttct
tggcttctct atgagtgaaa gtgtttcaaa ggtgtttttt 120tatgagacaa
cagtaattcc cctccttcag aacagagtgt agtgattggc agtgaagcta
180tccctctttg ccacccaatt ttgacggcag ttctcatagc atctcaaagc
aatagcagtg 240caaaagtaca taaccgtagg aaggtacgcg gtaggtattt
gagttcgttg gtggttatcc 300tccgcaaggc gcttcggcgg ttatttgttg
atagtcgaag aacaccaaaa aaaatgctgt 360tattgctttc tccgtaaaca
ataaaacccg gtagcgggat aacgcggctg atgcttttat 420ttaggaagga
atacttacat tatcatgaga acattgtcaa gggcattctg atacgggcct
480tccatcgcaa gaaaaaggca gcaacggact gagggacgga gagagttacg
gcataagaag 540tagtaggaga gcagagtgtc ataaagttat attattctcg
tcctaaagtc aattagttct 600gttgcgcttg acaatatatg tcgtgtaata
ccgtccctta gcagaagaaa gaaagacgga 660tccatatatg ttaaaatgct
tcagagatgt ttctttaatg tgccgtccaa caaaggtatc 720ttctgtagct
tcctctattt tcgatcagat ctcatagtga gaaggcgcaa ttcagtagtt
780aaaagcgggg aacagtgtga atccggagac ggcaagattg cccggccctt
tttgcggaaa 840agataaaaca agatatattg cactttttcc accaagaaaa
acaggaagtg gattaaaaaa 900tcaacaaagt ataacgccta ttgtcccaat
aagcgtcggt tgttcttctt tattatttta 960ccaagtacgc tcgagggtac
attctaatgc attaaaagac atg 1003561003DNASaccharomyces cerevisiae
56ataattgcgt ttaaacgcct ttccttttct tcatccattt tcctggtatt tatctcatcg
60tctctcttaa ttcttaattt actaccttca tctgggatta tgaactgtgg atgttccata
120tcacattcat ccttccctaa aggacaaaat ccggtcatgt atctttgaca
gaaaaccttc 180ttaatatgcc gtctaggaca agaactcccc agaggacaga
atcccatttc gtaattttca 240cattttggta tcttgctagc gggatctatg
tgtagatatt gacaatctgg actttgtgta 300cagtacccgt ttttgctgaa
gaagacacat tcaggcattt ttcgaagatt gtattcatgt 360aagtattcac
attggtcatt ctttttgcac aaccctcgaa gccaatgtct acaaacaatt
420ttattctgaa atattggtaa cacatgcttt tttggacata acggtcccct
cggacatgat 480ttagggcctt ctctagaatt gtaaaattca caaataggtc
tgtcaggatc gagtgaaaac 540gaatactctt gcctgaggaa aggttcaaat
ttaaaaggat attttgctgt atcggggtga 600attaggctca ttacataaac
ttattgattt tattcgtcct agaatacttg tttaagatac 660agtgcagaaa
tctttagctt ataattcaga aatgtgatat tttgaagcga ttggaacaac
720aaagtacagt gcattctata taattattaa aagttatagt ggactattgt
aataacactt 780gttttcccca tatcataacc gacggcgcgt aaatgcatgc
tcaatcaagt ttccgaataa 840gttaagtttg ccacccattg aggattaata
tacacaaggc agcattgaaa aacctaattg 900ttaacactac acaaatctct
attgttgtag taaactgcgc tttttaccac tgttacaata 960gttactaata
cacgcaaaaa ttctggataa acacggcaaa atg 1003571003DNASaccharomyces
cerevisiae 57gcacgtgaat gtgatgtctt gtgatggcgg gtgaattagg accggggtcc
gctgagtgtt 60gaacttcaca aagcatatat cattgacata atcagaaacg tgcttgatca
attgtgctat 120ggggaggtcc cggtgcaaga ttccaatcat ccttttgttc
gtgtcacctt tgttctgtgc 180catcaacgac aaatagtgcg tcacaagcag
cttcatcgtg attctctcct tcaggtggaa 240gttcaagaac tgcgaaattt
gaaactttgg gtaacacgat tggatttcct gaagaccttt 300tgccaatacc
acaatggcgt cctcgtggtc gtccagcagt tctgtgaact tggcctgtat
360cttgggagga ttgtgcaact cgtacgggta tgcgattgat agcaacgttt
gcaagctttt 420caaatacaaa gagttggttc tctcaatatg agggttgatt
accgcattat acggtagcct 480ctgaatggca ttaagccgtt tacacgtcaa
tgaaagcaac aggttgattg tcttgatcgt 540cagcatgtac tcctccttct
tggtcagcgg cggcctgtat tgcaggaaat actcgtagtt 600taggggcgca
attggtttac tagcgtagtc ctgtatcagc agctcaatgt tcgacctaat
660cttgtaatgc tggtcgaacg acagttgcga cagcagctcg tgtgagggtc
gctggcgatg 720tgcccagcgc attcccccac atttccatga acgcataatc
ttccacatac taccgtgaag 780gcgtcttgat accacgtctt ccagttatgg
tcttgactca ccaatgttct cagtttaatg 840ttgtttgctt catactggca
gacattttcc cttattaccg tgagaagcga gcggtggatt 900aatcgggatg
tcaaaaacaa gaaaattgca accttcttca cctataacct atatgatttg
960taggcagagg agtggaataa gcaacaaatc atagtcatca atg
1003581003DNASaccharomyces cerevisiae 58aaatctcact tcagaaacga
aaaatactac atagatatca ccgaattgtt ccatgaggtc 60accttccaaa ccgaattggg
ccaattgatg gacttaatca ctgcacctga agacaaagtc 120gacttgagta
agttctccct aaagaagcac tccttcatag ttactttcaa gactgcttac
180tattctttct acttgcctgt cgcattggcc atgtacgttg ccggtatcac
ggatgaaaag 240gatttgaaac aagccagaga tgtcttgatt ccattgggtg
aatacttcca aattcaagat 300gactacttag actgcttcgg taccccagaa
cagatcggta agatcggtac agatatccaa 360gataacaaat gttcttgggt
aatcaacaag gcattggaac ttgcttccgc agaacaaaga 420aagactttag
acgaaaatta cggtaagaag gactcagtcg cagaagccaa atgcaaaaag
480attttcaatg acttgaaaat tgaacagcta taccacgaat atgaagagtc
tattgccaag 540gatttgaagg ccaaaatttc tcaggtcgat gagtctcgtg
gcttcaaagc tgatgtctta 600actgcgttct tgaacaaagt ttacaagaga
agcaaataga actaacgcta atcgataaaa 660cattagattt caaactagat
aaggaccatg tataagaact atatacttcc aatataatat 720agtataagct
ttaagatagt atctctcgat ctaccgttcc acgtgactag tccaaggatt
780ttttttaagc caatgaaaat gaagaaatgc gtgatcggaa attacgggta
gtacgagaag 840gaaacttgag ccacccccca aattttattc atataataat
aggaaaagca acgacctcat 900ctctcgaaca ttgtttactt gagcaagtcc
gattaagagt aagttgtcgt acgttaaata 960caaataatca acaaaacact
acacaaaaac ttctacgata atg 1003591003DNASaccharomyces cerevisiae
59aaaaaactga aatatagggt ttgttaatcc tggtaaaaat ctgatgtgac atcttctgta
60cttaggtggt cttaccatga agtcagcatc gatacaattg atgttatatt tcgttggttc
120caatatgtaa attgcctcga tggaggattg tccttttctt gttggggaat
caatcaaatc 180aacagatgtg acattattta aaagttcttg gggtgtgagg
aaaagataac tcaatattgt 240ctccacagtt ttatcgataa taagaaactt
caagttattt ttcgtctcaa tctgattcaa 300aacccctatc aaatagttcc
tctgtaattc aattaaatca gacatcgttc cgccaaacca 360agcatgtgca
cactcttcta ctttgtcagt ggattctaat tattttcaag gtcttacaaa
420gcaatgtctt actagtttca atagggactt aaactgatcg tttaagcaaa
agaaaaacac 480cgtgctgtgt gaccatctac caataattgc gagaacataa
gacgacgccg ccattcacct 540ctctatcact tacaccatgt tatagggata
agtgtgtagt cgatcggcta ttcgatgatt 600tttaaagagt gcctgtaata
atagctgtat cgcaagtaag gaatacaaga ttactgatat 660tttgatccag
gaggatggac atgttagata gaaaaactaa gagggctact acatccggtt
720tagtattctg ttattgggaa tgcttttccc aagacgcaag caagaggtga
cacataaaca 780tagttttctt attgtatcgt tacatattct tattttacgc
tgctgtcatt aaatatggca 840cttatctcag atacatttct tttaatcagc
gaaaaaaatg gcgaggtgaa aaaaatcgat 900gagatgaaaa aattttttac
tagtaaaggc gtattgatga tatattagca gctaaagtat 960agcaaggttt
gtcaatttga agcagtccgc gtgttttaca atg 1003601003DNASaccharomyces
cerevisiae 60ctaactataa taactggtag ctttgtcact cgtaccagga aaagtgaaga
ttaaactgaa 60ttttaaaatg aacgatcgat tagttacgga agagcaagag ttgtttacaa
aattgcgcga 120gattgtaggt tcaagtattc gcttttggga ggaacaactg
ttttatcaag ttcaagatgt 180aagcaccata gaaaaccacg tcattctcag
tttaaaatgt acaattttaa cggatgctca 240gataagtacg ttcataagca
aacccagaga gcttcatacg catgccaaag gatatcctga 300aatctattac
ctttccgagt tatcaacaac tgtcaatttt ttttctaaag agggaaacta
360tgtcgaaata agccaggtta ttcctcattt taatgaatat ttttcctctt
taatagtgtc 420tcaattggaa tttgaatacc cgatggtctt ctccatgatt
tcaaggctcc gattgaagtg 480gcaacaaagt tcgctcgctc cgatatccta
cgccctaacg agcaattcag tacttcttcc 540aataatgctt aacatgattg
cccaagacaa atcttcaaca accgcgtatc aaattctgtg 600tcgaagaaga
ggtcctccaa ttcagaattt tcaaattttt tccttaccgg ctgtaacgta
660caataagtag catgcataaa atataattta atcaaatact tttgggcaat
taaaatttta 720gttaacaata gttatgcaat gcgctttatg ttcatatgat
accgtttata agctattgcc 780atatccttat cttattgctt ccagtagcct
cgagtcgacc actaaaaaga tgtcacttaa 840gacggaaatt atgtagctgc
acttcttttt taacaagttc ggtcggccct tcaagttctc 900ctttctaaag
cctcattatt tattgcgtag atgctaaatg ttatcgcggt ttagcttgca
960tgttacgttt ccgttttaga acctggtcga gtagcgaata atg
1003611003DNASaccharomyces cerevisiae 61ttcgccaacc atcatacccg
aatgtttcat tagcgaactg aatgagagca tgccctggta 60tcagcttaga taagttgtgc
ttcccatatt tattattgtg gtaaatattg tacgtgtacc 120tttcgatctt
ggactgtaga agcccaatcc tctgcaccga ccaagcgctg gcaggtcttc
180cattccaatc ttcgatgatt tggaactctt ttatatccaa gcccggtgct
gtgccgtatg 240tagtcgaatc atacgatatg ctcgaactgg gcttcgcttg
tatcgtcatt attttgtctg 300tttatcctga aatatgctct tctcctctac
gtaacccttt aatggtccaa agcatgtgct 360aaactgccac agactccctc
aatgatgttc ttatccttcc ttgccgttac tgcttcctat 420catatctgcc
cttgccaaac tgtggcttcc tctaatcaat ttgccccagt acattgaaaa
480gcacgtgacg tgacaccaac ttgaaagaca ttagtgtcca acgcgatata
atggtcaata 540aaatacccaa ttttatcaag tgaacatgat attattcaat
actatagtac tttaaagaaa 600acttcaacaa gttgcttcat tgctatatta
tacaattatg ggtgatgccg gtcggctaac 660ctttctttcg cagcgccaca
tacaagaaca tatggtaatt actaagaact ttttggaaac 720tatttccatg
aagaaaacaa aatttactgg cttctgtctt cgaagtaaat ttagcgaact
780tgatactttc gtttggacag tcgtttagat ttgcaactgt ctgttcttat
ttcttgctat 840tattaccaga aataaagcct aaagcgcgtg gttcttacga
aaaaaataga ccttgcacat 900taacagggtt tagacatcac agagactgaa
tactggagag aagacaattc ttgttcacga 960cttcgagtga ttaatatcta
taacaaccct cctatcaaga atg 1003621003DNASaccharomyces cerevisiae
62tgaaaggcaa atttacgctc ttcaagacac caatttagga ttggtggcta ccgcggagat
60acctttggca ggtctagggg caaacaaagt tctagagtta aactcgggag aatgttctaa
120gaaactagtt ggagtaagta gatgttacag agctgaggca ggtgccaggg
gaaaagatac 180gaaaggtctc tatcgcgttc atgagtttac taaagtagaa
ttattttgct ggagcaagcc 240agaaaccagt gcaaaggtcc ttgaggaaat
aaagcaattt cagatttctg tagttgagga 300attaggaata ccggccaaag
tactaaatat gccatcaaac gatcttggta atcccgcttt 360caagaaatac
gacatcgaag cttggatgcc aggaaggggg aaatttggcg agataagtag
420tgcctccaat tgtaccgatt tccaaagtag aagattaaat acaaagtaca
gggacgataa 480cacaggaaag ctagaatatg tgcacacgct gaatggcaca
gcaatggcta tcccaagagt 540gatagtagca ctagtagaaa atttctatga
cccaagcacc ggtaagatat ctgttcctga 600atgtttgagg gagtttatga
atggccaacg atacatttaa taagtaagaa aatcttgtaa 660atatgtaatc
ttatgttaac cgtagcatca ttgtgttgga cgaaatcctc tgtgatgttc
720tgtgtgattg aaattgtatg gtattttcga tcatacgagt atttttccta
aagatgaaag 780agaactcgat tcctaaacag ttgtttgtag ggcataacgt
tgtatcgtaa atacttacca 840tacgcttagg tatattcctt tagatttccg
taacaccttt cagttttact attccgcgtg 900gcgaggtaaa gatcataaaa
atacaagttt gttatagcat ttcctgcaaa taattgctgt 960aactagtggc
gaaaaggtca tagaattatt cgcctaaatt atg 1003631003DNASaccharomyces
cerevisiae 63ccggaagtaa ttttatcaat tatagtttcc aaatcatcat attcttgaac
tatatcaatt 60aaatcatcac tcgtccagtc agggaacaat tctgtaagcg tatctatttt
ggactttagc 120gcaggattta gttttttact actatgacta ttatgattag
acttcctaaa ttgtgtagac 180attatatata tatgatgatg tagattatat
tgagaaaacg accaaacaaa aataaatatg 240taagtacttt atgatggtga
tttttttggg gtaattggta agagtttttc ttgtagcttc 300aaaagatcaa
atataaaata cgaatgtttc tgaaattact gtaaaagact taggaaaagg
360aatgaaacgt ttatatggga gagaacgaaa atttttcctt ttcccgccac
tactttttgc 420caccatttcc cgttatttat ttgggcgagt gctcacctcc
gacggaatca cgggtgacgg 480atatcctaaa aaggtaaaga attgaaacag
atctattaat atacggatca ctatacatta 540ttataagtcg atattgcagt
attagtattt acatgttgaa taatgcacat tctgtgctaa 600aatatatact
tcatttgtca acttctttta gtttgccgcc attttcaaac ggcgcctcct
660ccctagcttc gtctagagca ggcttaatac cttgtttgac gagagctgcg
ttgacacatg 720tagtataagc gtaccattgt tttgagcact cgttctcaac
ggattttccc ttcaggaatt 780tttcgctata ccattcatta aaacaactat
cgtatttcgt cttcaggtca gtgcattcag 840gcgcaaaact agctgacatt
atattcccca tataaaactg gtattattgt tattgtaatt 900caagttaaaa
cacaaacaga tgttttctaa tgacctctcc cggcatatat ctggcgcact
960tagaataaat ttctacatgc aaatactctg aatattctga atg
1003641003DNASaccharomyces cerevisiae 64ttcttcttct ccattattat
cttcagtagt tgtcttgtca tcagattgtc cctgttgcga 60tttttcatca tctgaatgtt
ctggatcttt tttttcagaa gatccagcat tgttgtctag 120tgttttgtca
ctagtatttt cttccgagtt ctcagaactt atgttttcat ccttttgatt
180gttagcggtg gcattgtctt cattggattg ctcatcatct ttggttgata
cctttgattc 240atcagtatct ttatcttcgg aagacttgtt gtcttcttct
gctagaacaa ccaattcacc 300atcttcagct acttttgaag caccaatttt
atcaccccct aattgagggc cagcttcagt 360ttcatcgttt ttgctgtcgt
tgtcatctcc attttcatcg cttttgctgt tgttgtcatc 420ttcattttct
ggtgattgct caccaccact ggaaagctct tcctctgcgg ttttatcgga
480ttctaccttc cttgaggcga aaagaggctt cctattagga gcaataaaat
ataaagcacc 540agccatagaa agaatcccca ttataaagcc cgctgttttt
tcctgattgg agttcctacc 600gaactgaggg gaggacgcca tgagacgtct
tgtttggtgt cggcataacc cccttgccac 660ttgaattgac ggcctgtttc
tgcacgcatt cctgacgact aagttgcgaa gcattttact 720gataatatac
actctttgga tcgagcctac ttccagttgg taattggtgt tccacaattt
780cagcattata tgtttttaaa ccaaaattcg gctccttttc cctttttttc
ttattgggtg 840gcgtgccgta cagaacgatt ggcttggtgt gaaatcaaga
gcaagcacaa tagatatcaa 900catgaacaat atacaaaagt ctctggcaca
gtttgactgc gttagaccag gctagggcat 960ttctgaagct ttacgtatca
ctagagaagt tattttggca atg 1003651003DNASaccharomyces cerevisiae
65gaaagatgac ctctttcaaa cgcagtacgc agtttgttga ccatatgcgg ataggtgacg
60tatattttcc agaatggtaa tttggtatct ccggaataac cccagataga ctgtttagat
120acaacttcaa tcgaatcaat agcgtggtca aatgtttcgt ttaaatagtg
cacaaacttg 180ttgatttgct cctgatcgtt agcgtcggaa gaattgggcg
ctgggacgta aagatagttc 240ttgaaccctg taacattaca aagtacagag
tgtccttcac tagtgacacc aaaaaacctt 300accacggtag atgtattttc
atctttgata ccattcagta cgctctgttc cgcatcaatt 360tgttggaaag
aaatatcata caaacttggg tcaaaatcgg ttggaagttt cttacgctcg
420aatgatgaca ggtcatgctc ttcttggtcg gccatatcat gttccatttg
cgatacgtct 480tgttcaaaag tagactctaa ttgcgtaccc attaaatctg
tatcctttgc tttgaagcct 540tgactattat attttcgaaa agaatcactc
ggaataatct ctattgttga aacaggttca 600cttccaacac catgatctat
tgattgccgt ttgattttct tttccaactg gggagtatcc 660tcgtcatcga
tcttcacatc aaccatggga agggatcttt tttcactcat tgctgtgcgt
720atatgtataa agattaatgc ttaatagcaa gtgctcaata tctattttcc
tttatccttg 780aaaattgcct tagatcagta gtactgagtg tttgttcttc
ataagcgaat agcagatgtc 840ctcattgaac attttataga ataaaaagtt
acgcgtaata cgaaaaattg cctattacgt 900taaggcatac ttttaccaca
caaaaccaag atcaaccgtc aagcaagaac gctgcaaaga 960aacgcagata
taaaggagaa caccagatcg aactgcaact atg 1003661003DNASaccharomyces
cerevisiae 66tcttgaatgt cgacatatgc ccatcatcag tgaggaatac tcgcgtgaag
gatctcatct 60gcgacctgag tgatgatgaa gaagttgcag cactactgaa tttgttgaaa
agaaagtata 120aaaatgaaat ccgattgatt gtgaacaatg ctggagtaag
ggccaacttc accggattca 180acggcatgga acgggataat cttgataaga
tttttaagat aaacacattt gcaccacttc 240agttcattca agaactggcc
cctagtagac actcaactag acagtgttat attgtcaata 300tcgcaagcat
tttgggcata ttgacgccgg ccaaagtagc agcttatgcg gcgagcaaag
360cagcattgat agcctttcat cagtcgtaca gttttgaatt gcaaaacgaa
ggcgtaagaa 420atatcagaac actattagtg accccagggc aactgaatac
ggaaatgttt gcaggattca 480agcctcctcg ccagttcttt gcccctgtaa
tagacattac tactctagct gccaagatag 540tccgttattg tgagctgggt
cagaggggac agctaaatga acccttttat tgtagttttg
600ctcatctttt gatgtgcgta ccttattcgc tacaacgcat tgtaagaagc
ttctctcgca 660tagattgctg cctcccggac gagtagaaag ctatacatag
taaatagaat atttatagat 720gaatccgagt agacagcctt ggtgtttgct
tatacattct ttgcaattat gcacgccctc 780attaccgttg ctcatatttt
tgggcattaa ttgtattttg aaaagtgctc gttcaggccc 840gtgtaaaaaa
aggtgatgaa gcaaacatta taataaacac ttctgagaag ccacgtgtag
900aggggtaagc tatatcgtaa acaccgttgg atgtggacca cggtgcactg
ataaaataaa 960gatatagtag tagaatctgt tactaatctt aacacttttg atg
1003671003DNASaccharomyces cerevisiae 67gctgtagata gatatagaga
agccgccaaa acagaactaa gaattctaca gactatcctg 60aataatgacc ctcaaggtca
gttccagtgc ctcttgctaa gggagtgctt cgattacaaa 120aatcacattt
gtttggtgac agatctatac ggcaggtcca tttacgattt tatgtgctcc
180aacggcattg ccaggttccc cggctctcat attcaggcca ttgccagaca
gctaatcaga 240tctgtctgct tcttgcacga tttgggcata atacacacgg
atttgaaacc agaaaatatc 300ctgatttgtg acgaaaccca tattgctcaa
aaattgcctt tgaaaaccgt acagtcgcta 360agcaagagac gccgtgaagc
cagtaagggg aagcgcaaaa tcttgaaaaa tccagaaatc 420aaaatcattg
atttcggtag cgcaattttc cattacgaat atcatcctcc tgtaatatcc
480actcgtcact atagagcccc ggaaattgtc cttggcttgg gctggtcgtt
cccctgcgac 540atttggtcca tcgcgtgtgt cctagtagaa ctggttattg
gcgaatctct ttaccccatt 600catgaaaatc tagaacatat ggctatgatg
caacgaatca atggaacccc tttccccaca 660gacattattg ataagatgtt
ttacaaatct aaacataaat tgggcaactc tccatcagac 720ctaaattcaa
cggtgataaa gcatttcgac agaaaaactt taagcttaca atggcctgaa
780aaaaataaac gcggagatac cataaccacc gaaaagtcaa tgaaaagggt
cttgcagtca 840tgtgatcgct tagatattta catctcaaag gtcttgaaac
aggattacgg cgatagctta 900agcatcaatt ggaatctacc gccggagaaa
aactggtctt tgataaattc gaaattggca 960tggaaaagac aaacacattc
ctcttcttcc tcaactactg atg 100368987DNASaccharomyces cerevisiae
68cggtccagaa aagcaactga gattgggagt ataattcggt ggcgttattg gaatacataa
60taggaaaata cggcagatac ctagtcacga aaatatggtg taatctattc gctttttcaa
120tggaaatact aatatcgccc agtacaaact catctacatt tgcatgcggc
gaaggcaata 180atggcatagt cgtggtggtg gctactaccg gcgtcttcgt
cctatctgca ttgttatttg 240ttgctacgtg gcttgttgct gcaaatggtg
gttgatttgt tgtcgttgta acggtggtcg 300acgtagtatg aggggaatat
gttttttgaa ttggagagaa ggggccgttc ggagtgttac 360ctgcagagtt
gttcttataa aaagccattt gcaaagcagg aggtaaagat tcttttgaat
420tattaggcaa tgctgagtca tttagcagcg ggggcagctt atttggctct
gttgcaacca 480agcctttcga atcttgcgct aaagaagatg cgcgcaacgt
catgtgagaa gatgcttcat 540tatttgcttt aaatttattc gtattgctgc
cctgatttgc ttgtaagagt tggggttccc 600tggacaaata agtttgaact
gaaaccttag aatctcgttg agccgcggaa gaagttggag 660aacctgagga
aggagaagaa tctgggttag ggataatagt acccggagtt ggagttggat
720gcagattgag cttattcaaa aggctattgc ccatgggaat ctgttgtaaa
agatgaacga 780aaacgctgtc attggccaga agagtatcga gtttagattt
gatttcatcc acatcttgtc 840tcagtagttg caactgtgag cccttcttag
gcctgaattg aggattgatt tcacagtgga 900gaccaatttt ttcgcatctg
gagcaaggat gagggaaatt ttgactagca tcgcatttga 960ttttgtgctg
tctacaatgt gtacatg 9876930DNAArtificial SequencePrimer 69tttgcgcgcg
ggccgcatca tgtaattagt 307030DNAArtificial SequencePrimer
70aaaagatctg accgagcgca gcgagtcagt 307183DNAArtificial
SequencePrimer 71cccatgttaa ttaaatctgt tgtttattca attttagccg
cttctttggc caatgcaggt 60gcttccaagg tgtacgaccc cga
837232DNAArtificial SequencePrimer 72aaaggcgcgc cttactgctc
gttcttcagc ac 327319PRTArtificial SequencePHO5 signal sequence
73Met Leu Ile Lys Ser Val Val Tyr Ser Ile Leu Ala Ala Ser Leu Ala1
5 10 15Asn Ala Gly7431DNAArtificial SequencePrimer 74tttggatcca
cggattagaa gccgccgagc g 317535DNAArtificial SequencePrimer
75cccttaatta acatggtttt ttctccttga cgtta 357634DNAArtificial
SequencePrimer 76tttggatcct gtgcacgatg gcttcgtttt agcc
347733DNAArtificial SequencePrimer 77cttaattaac atcgagtttc
aaaaaattgt aag 337832DNAArtificial SequencePrimer 78cccggatcct
ctacgatata tcctgtaaat ag 327934DNAArtificial SequencePrimer
79cttaattaac attttgagta cgtctaatct gtat 348084DNAArtificial
SequencePrimer 80cccatgggat ccaaatctgt tgtttattca attttagccg
cttctttggc caatgcaggt 60gaagacgcca aaaacataaa gaaa
848134DNAArtificial SequencePrimer 81aaaggcgcgc cttactttcc
gcccttcttg gcct 348219PRTArtificial SequencePHO5 signal sequence
82Met Leu Ile Lys Ser Val Val Tyr Ser Ile Leu Ala Ala Ser Leu Ala1
5 10 15Asn Ala Gly8331DNAArtificial SequencePrimer 83tttggatcca
cggattagaa gccgccgagc g 318433DNAArtificial SequencePrimer
84tttggatccc atggtttttt ctccttgacg tta 33857393DNAArtificial
SequencepSPRT47 85gtacgctgca ggtcgacgga tccccgggtt aattaaatct
gttgtttatt caattttagc 60cgcttctttg gccaatgcag gtgcttccaa ggtgtacgac
cccgagcaac gcaaacgcat 120gatcactggg cctcagtggt gggctcgctg
caagcaaatg aacgtgctgg actccttcat 180caactactat gattccgaga
agcacgccga gaacgccgtg atttttctgc atggtaacgc 240tgcctccagc
tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg ctagatgcat
300catccctgat ctgatcggaa tgggtaagtc cggcaagagc gggaatggct
catatcgcct 360cctggatcac tacaagtacc tcaccgcttg gttcgagctg
ctgaaccttc caaagaaaat 420catctttgtg ggccacgact ggggggcttg
tctggccttt cactactcct acgagcacca 480agacaagatc aaggccatcg
tccatgctga gagtgtcgtg gacgtgatcg agtcctggga 540cgagtggcct
gacatcgagg aggatatcgc cctgatcaag agcgaagagg gcgagaaaat
600ggtgcttgag aataacttct tcgtcgagac catgctccca agcaagatca
tgcggaaact 660ggagcctgag gagttcgctg cctacctgga gccattcaag
gagaagggcg aggttagacg 720gcctaccctc tcctggcctc gcgagatccc
tctcgttaag ggaggcaagc ccgacgtcgt 780ccagattgtc cgcaactaca
acgcctacct tcgggccagc gacgatctgc ctaagatgtt 840catcgagtcc
gaccctgggt tcttttccaa cgctattgtc gagggagcta agaagttccc
900taacaccgag ttcgtgaagg tgaagggcct ccacttcagc caggaggacg
ctccagatga 960aatgggtaag tacatcaaga gcttcgtgga gcgcgtgctg
aagaacgagc agtaaggcgc 1020gcgggccgca tcatgtaatt agttatgtca
cgcttacatt cacgccctcc ccccacatcc 1080gctctaaccg aaaaggaagg
agttagacaa cctgaagtct aggtccctat ttattttttt 1140atagttatgt
tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
1200agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
tttgggacgc 1260tcgaaggctt taatttgcgg ccctgcatta atgaatcggc
caacgcgcgg ggagaggcgg 1320tttgcgtatt gggcgctctt ccgcttcctc
gctcactgac tcgctgcgct cggtcagatc 1380tgtttagctt gcctcgtccc
cgccgggtca cccggccagc gacatggagg cccagaatac 1440cctccttgac
agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
1500ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
gccgcacggc 1560gcgaagcaaa aattacggct cctcgctgca gacctgcgag
cagggaaacg ctcccctcac 1620agacgcgttg aattgtcccc acgccgcgcc
cctgtagaga aatataaaag gttaggattt 1680gccactgagg ttcttctttc
atatacttcc ttttaaaatc ttgctaggat acagttctca 1740catcacatcc
gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga
1800ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
taatgtcggg 1860caatcaggtg cgacaatcta tcgattgtat gggaagcccg
atgcgccaga gttgtttctg 1920aaacatggca aaggtagcgt tgccaatgat
gttacagatg agatggtcag actaaactgg 1980ctgacggaat ttatgcctct
tccgaccatc aagcatttta tccgtactcc tgatgatgca 2040tggttactca
ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct
2100gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
gcattcgatt 2160cctgtttgta attgtccttt taacagcgat cgcgtatttc
gtctcgctca ggcgcaatca 2220cgaatgaata acggtttggt tgatgcgagt
gattttgatg acgagcgtaa tggctggcct 2280gttgaacaag tctggaaaga
aatgcataag cttttgccat tctcaccgga ttcagtcgtc 2340actcatggtg
atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
2400attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
cctatggaac 2460tgcctcggtg agttttctcc ttcattacag aaacggcttt
ttcaaaaata tggtattgat 2520aatcctgata tgaataaatt gcagtttcat
ttgatgctcg atgagttttt ctaatcagta 2580ctgacaataa aaagattctt
gttttcaaga acttgtcatt tgtatagttt ttttatattg 2640tagttgttct
attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
2700tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
cgtatgtgaa 2760tgctggtcgc tatactgctg tcgattcgat actaacgccg
ccatccagtg tcgaaaacga 2820gctcgaattc ctgggggaac aacttcacag
aatgttttgt catattgtcg aagtggtcac 2880aaaacaagag aagttccgcc
aattataaaa agggaacccg tatatttcag cttcacggat 2940gatttccagg
gtgagagtac tgtatatggg cttacgatag aaggccataa aaatttcttg
3000cttggcaaca aaatagaagt gaaatcatgt cgaggctgct gtgtgggaga
acagcttaaa 3060atatcacaaa aaaagaatct aaaacactgt gttgcttgtc
ccagaaaggg aatcaagtat 3120ttttataaag attggagtgg taaaaatcga
gtatgtgcta gatgctatgg aagatacaaa 3180ttcagcggtc atcactgtat
aaattgcaag tatgtaccag aagcacgtga agtgaaaaag 3240gcaaaagaca
aaggcgaaaa attgggcatt acgcccgaag gtttgccagt taaaggacca
3300gagtgtataa aatgtggcgg aatcttacag tggcctatgc ggccgctcta
gaactagtgg 3360atcgatcccc aattcgccct atagtgagtc gtattacaat
tcactggccg tcgttttaca 3420acgtcgtgac tgggaaaacc ctggcgttac
ccaacttaat cgccttgcag cacatccccc 3480tttcgccagc tggcgtaata
gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 3540cagcctgaat
ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
3600ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
attaagcgcg 3660gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
ccagcgccct agcgcccgct 3720cctttcgctt tcttcccttc ctttctcgcc
acgttcgccg gctttccccg tcaagctcta 3780aatcggggtg ggccatcgcc
ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3840ttctttaata
gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
3900tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
tgagctgatt 3960taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
ttacaatttt atggtgcact 4020ctcagtacaa tctgctctga tgccgcatag
ttaagccagc cccgacaccc gccaacaccc 4080gctgacgcgc cctgacgggc
ttgtctgctc ccggcatccg cttacagaca agctgtgacc 4140gtctccggga
gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
4200aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
ggtttcttag 4260acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
ctatttgttt atttttctaa 4320atacattcaa atatgtatcc gctcatgaga
caataaccct gataaatgct tcaataatat 4380tgaaaaagga agagtatgag
tattcaacat ttccgtgtcg cccttattcc cttttttgcg 4440gcattttgcc
ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
4500gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
taagatcctt 4560gagagttttc gccccgaaga acgttttcca atgatgagca
cttttaaagt tctgctatgt 4620ggcgcggtat tatcccgtat tgacgccggg
caagagcaac tcggtcgccg catacactat 4680tctcagaatg acttggttga
gtactcacca gtcacagaaa agcatcttac ggatggcatg 4740acagtaagag
aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
4800cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
catgggggat 4860catgtaactc gccttgatcg ttgggaaccg gagctgaatg
aagccatacc aaacgacgag 4920cgtgacacca cgatgcctgt agcaatggca
acaacgttgc gcaaactatt aactggcgaa 4980ctacttactc tagcttcccg
gcaacaatta atagactgga tggaggcgga taaagttgca 5040ggaccacttc
tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
5100ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
gccctcccgt 5160atcgtagtta tctacacgac ggggagtcag gcaactatgg
atgaacgaaa tagacagatc 5220gctgagatag gtgcctcact gattaagcat
tggtaactgt cagaccaagt ttactcatat 5280atactttaga ttgatttaaa
acttcatttt taatttaaaa ggatctaggt gaagatcctt 5340tttgataatc
tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
5400cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
aatctgctgc 5460ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
tgccggatca agagctacca 5520actctttttc cgaaggtaac tggcttcagc
agagcgcaga taccaaatac tgtccttcta 5580gtgtagccgt agttaggcca
ccacttcaag aactctgtag caccgcctac atacctcgct 5640ctgctaatcc
tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
5700gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
gggttcgtgc 5760acacagccca gcttggagcg aacgacctac accgaactga
gatacctaca gcgtgagcat 5820tgagaaagcg ccacgcttcc cgaagggaga
aaggcggaca ggtatccggt aagcggcagg 5880gtcggaacag gagagcgcac
gagggagctt ccagggggaa acgcctggta tctttatagt 5940cctgtcgggt
ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
6000cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
cttttgctgg 6060ccttttgctc acatgttctt tcctgcgtta tcccctgatt
ctgtggataa ccgtattacc 6120gcctttgagt gagctgatac cgctcgccgc
agccgaacga ccgagcgcag cgagtcagtg 6180agcgaggaag cggaagagcg
cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 6240cattaatgca
gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
6300attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
gcttccgcgg 6360ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
cacaggaaac agctatgacc 6420atgattacgc cactagtccg aggcctcgag
atccgatatc gccgtggcgg ccgccagctg 6480aagcttaatt atcctgggca
cgagtgaaac aaagctaaaa cctttattta gcatggccat 6540tgaatgtaac
aattatatat atcgcaagca caaaaaatca aggagagaga actaccactt
6600tgttcatgtg tacaatgttc attatctcca taagcaaaaa aaaaaaatag
aaaacatatg 6660ctataaggtt gatattctca cgagtaagcg gcacttgcta
cttattgaca ttgcagattt 6720ttggctacag aaatagtata ttagagatta
taattgctaa tcaaatcaaa atataaaatt 6780agtaaaccaa accatttata
cccttcctta gtagttatgg attgtttttt aatgatattt 6840ctgcaaacca
aagaaagatt gttatccaga tagaatttag ttttgatatt catttttttg
6900ttgaagattg aacgccatat ctgggcctca taattcaaaa gacggtgcca
ttatcggtag 6960cgtttcgcat tgtactggat ttcagaaatt tcacagttga
tgaatcgaaa agaatggtct 7020cattgcaaca cgtaaggtta agatgtccct
ttttaccatt ataggcaata aatgaatcat 7080aaaacgaccg tatactggtg
aaatagtagg gagaacgagt acctgtagta aaaagtataa 7140atcatagtta
atcgggcaat gtccctcgat caaggagtat tgtgtcatgt tcgagacaaa
7200cgccaacatt tttgtttctt ttggacaaat gttgtttgca tttatgatcc
gttatatttt 7260gatctaatgt agagttgcac gtagttctta ctggcaaaga
aatcgatgca taccaaaaaa 7320gaataaaggt gatatttgat ctttaccgtt
tagttccaac gtaaaattgt gcctttggac 7380ttaaaatggc gtc
7393868090DNAArtificial SequencepSPRT50 86gtacgctgca ggtcgacgga
tccaaatctg ttgtttattc aattttagcc gcttctttgg 60ccaatgcagg tgaagacgcc
aaaaacataa agaaaggccc ggcgccattc tatcctctag 120aggatggaac
cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg
180gaacaattgc ttttacagat gcacatatcg aggtgaacat cacgtacgcg
gaatacttcg 240aaatgtccgt tcggttggca gaagctatga aacgatatgg
gctgaataca aatcacagaa 300tcgtcgtatg cagtgaaaac tctcttcaat
tctttatgcc ggtgttgggc gcgttattta 360tcggagttgc agttgcgccc
gcgaacgaca tttataatga acgtgaattg ctcaacagta 420tgaacatttc
gcagcctacc gtagtgtttg tttccaaaaa ggggttgcaa aaaattttga
480acgtgcaaaa aaaattacca ataatccaga aaattattat catggattct
aaaacggatt 540accagggatt tcagtcgatg tacacgttcg tcacatctca
tctacctccc ggttttaatg 600aatacgattt tgtaccagag tcctttgatc
gtgacaaaac aattgcactg ataatgaatt 660cctctggatc tactgggtta
cctaagggtg tggcccttcc gcatagaact gcctgcgtca 720gattctcgca
tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt
780taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga
tatttgatat 840gtggatttcg agtcgtctta atgtatagat ttgaagaaga
gctgttttta cgatcccttc 900aggattacaa aattcaaagt gcgttgctag
taccaaccct attttcattc ttcgccaaaa 960gcactctgat tgacaaatac
gatttatcta atttacacga aattgcttct gggggcgcac 1020ctctttcgaa
agaagtcggg gaagcggttg caaaacgctt ccatcttcca gggatacgac
1080aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag
ggggatgata 1140aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
gaaggttgtg gatctggata 1200ccgggaaaac gctgggcgtt aatcagagag
gcgaattatg tgtcagagga cctatgatta 1260tgtccggtta tgtaaacaat
ccggaagcga ccaacgcctt gattgacaag gatggatggc 1320tacattctgg
agacatagct tactgggacg aagacgaaca cttcttcata gttgaccgct
1380tgaagtcttt aattaaatac aaaggatatc aggtggcccc cgctgaattg
gaatcgatat 1440tgttacaaca ccccaacatc ttcgacgcgg gcgtggcagg
tcttcccgac gatgacgccg 1500gtgaacttcc cgccgccgtt gttgttttgg
agcacggaaa gacgatgacg gaaaaagaga 1560tcgtggatta cgtcgccagt
caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt 1620ttgtggacga
agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga
1680tcctcataaa ggccaagaag ggcggaaagt aaggcgcgcg ggccgcatca
tgtaattagt 1740tatgtcacgc ttacattcac gccctccccc cacatccgct
ctaaccgaaa aggaaggagt 1800tagacaacct gaagtctagg tccctattta
tttttttata gttatgttag tattaagaac 1860gttatttata tttcaaattt
ttcttttttt tctgtacaga cgcgtgtacg catgtaacat 1920tatactgaaa
accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccc
1980tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
cgctcttccg 2040cttcctcgct cactgactcg ctgcgctcgg tcagatctgt
ttagcttgcc tcgtccccgc 2100cgggtcaccc ggccagcgac atggaggccc
agaataccct ccttgacagt cttgacgtgc 2160gcagctcagg ggcatgatgt
gactgtcgcc cgtacattta gcccatacat ccccatgtat 2220aatcatttgc
atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
2280cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
tgtccccacg 2340ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
actgaggttc ttctttcata 2400tacttccttt taaaatcttg ctaggataca
gttctcacat cacatccgaa cataaacaac 2460catgggtaag gaaaagactc
acgtttcgag gccgcgatta aattccaaca tggatgctga 2520tttatatggg
tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
2580attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
gtagcgttgc 2640caatgatgtt acagatgaga tggtcagact aaactggctg
acggaattta tgcctcttcc 2700gaccatcaag cattttatcc gtactcctga
tgatgcatgg ttactcacca ctgcgatccc 2760cggcaaaaca gcattccagg
tattagaaga atatcctgat tcaggtgaaa atattgttga 2820tgcgctggca
gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
2880cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
gtttggttga 2940tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
gaacaagtct ggaaagaaat 3000gcataagctt ttgccattct caccggattc
agtcgtcact catggtgatt tctcacttga 3060taaccttatt tttgacgagg
ggaaattaat aggttgtatt gatgttggac gagtcggaat 3120cgcagaccga
taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
3180attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
ataaattgca 3240gtttcatttg atgctcgatg agtttttcta atcagtactg
acaataaaaa gattcttgtt 3300ttcaagaact tgtcatttgt atagtttttt
tatattgtag ttgttctatt ttaatcaaat
3360gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
gaagttaagt 3420gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
tggtcgctat actgctgtcg 3480attcgatact aacgccgcca tccagtgtcg
aaaacgagct cgaattcctg ggggaacaac 3540ttcacagaat gttttgtcat
attgtcgaag tggtcacaaa acaagagaag ttccgccaat 3600tataaaaagg
gaacccgtat atttcagctt cacggatgat ttccagggtg agagtactgt
3660atatgggctt acgatagaag gccataaaaa tttcttgctt ggcaacaaaa
tagaagtgaa 3720atcatgtcga ggctgctgtg tgggagaaca gcttaaaata
tcacaaaaaa agaatctaaa 3780acactgtgtt gcttgtccca gaaagggaat
caagtatttt tataaagatt ggagtggtaa 3840aaatcgagta tgtgctagat
gctatggaag atacaaattc agcggtcatc actgtataaa 3900ttgcaagtat
gtaccagaag cacgtgaagt gaaaaaggca aaagacaaag gcgaaaaatt
3960gggcattacg cccgaaggtt tgccagttaa aggaccagag tgtataaaat
gtggcggaat 4020cttacagtgg cctatgcggc cgctctagaa ctagtggatc
gatccccaat tcgccctata 4080gtgagtcgta ttacaattca ctggccgtcg
ttttacaacg tcgtgactgg gaaaaccctg 4140gcgttaccca acttaatcgc
cttgcagcac atcccccttt cgccagctgg cgtaatagcg 4200aagaggcccg
caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
4260tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
cgtcaaagca 4320accatagtac gcgccctgta gcggcgcatt aagcgcggcg
ggtgtggtgg ttacgcgcag 4380cgtgaccgct acacttgcca gcgccctagc
gcccgctcct ttcgctttct tcccttcctt 4440tctcgccacg ttcgccggct
ttccccgtca agctctaaat cggggtgggc catcgccctg 4500atagacggtt
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
4560ccaaactgga acaacactca accctatctc gggctattct tttgatttat
aagggatttt 4620gccgatttcg gcctattggt taaaaaatga gctgatttaa
caaaaattta acgcgaattt 4680taacaaaata ttaacgttta caattttatg
gtgcactctc agtacaatct gctctgatgc 4740cgcatagtta agccagcccc
gacacccgcc aacacccgct gacgcgccct gacgggcttg 4800tctgctcccg
gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
4860gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
tacgcctatt 4920tttataggtt aatgtcatga taataatggt ttcttagacg
tcaggtggca cttttcgggg 4980aaatgtgcgc ggaaccccta tttgtttatt
tttctaaata cattcaaata tgtatccgct 5040catgagacaa taaccctgat
aaatgcttca ataatattga aaaaggaaga gtatgagtat 5100tcaacatttc
cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
5160tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
cacgagtggg 5220ttacatcgaa ctggatctca acagcggtaa gatccttgag
agttttcgcc ccgaagaacg 5280ttttccaatg atgagcactt ttaaagttct
gctatgtggc gcggtattat cccgtattga 5340cgccgggcaa gagcaactcg
gtcgccgcat acactattct cagaatgact tggttgagta 5400ctcaccagtc
acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
5460tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
tcggaggacc 5520gaaggagcta accgcttttt tgcacaacat gggggatcat
gtaactcgcc ttgatcgttg 5580ggaaccggag ctgaatgaag ccataccaaa
cgacgagcgt gacaccacga tgcctgtagc 5640aatggcaaca acgttgcgca
aactattaac tggcgaacta cttactctag cttcccggca 5700acaattaata
gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
5760tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
ctcgcggtat 5820cattgcagca ctggggccag atggtaagcc ctcccgtatc
gtagttatct acacgacggg 5880gagtcaggca actatggatg aacgaaatag
acagatcgct gagataggtg cctcactgat 5940taagcattgg taactgtcag
accaagttta ctcatatata ctttagattg atttaaaact 6000tcatttttaa
tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
6060cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
tcaaaggatc 6120ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
caaacaaaaa aaccaccgct 6180accagcggtg gtttgtttgc cggatcaaga
gctaccaact ctttttccga aggtaactgg 6240cttcagcaga gcgcagatac
caaatactgt ccttctagtg tagccgtagt taggccacca 6300cttcaagaac
tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
6360tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
agttaccgga 6420taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
cagcccagct tggagcgaac 6480gacctacacc gaactgagat acctacagcg
tgagcattga gaaagcgcca cgcttcccga 6540agggagaaag gcggacaggt
atccggtaag cggcagggtc ggaacaggag agcgcacgag 6600ggagcttcca
gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
6660acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
aaaacgccag 6720caacgcggcc tttttacggt tcctggcctt ttgctggcct
tttgctcaca tgttctttcc 6780tgcgttatcc cctgattctg tggataaccg
tattaccgcc tttgagtgag ctgataccgc 6840tcgccgcagc cgaacgaccg
agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 6900aatacgcaaa
ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
6960gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
agctcactca 7020ttaggcaccc caggctttac actttatgct tccgcggctc
gtatgttgtg tggaattgtg 7080agcggataac aatttcacac aggaaacagc
tatgaccatg attacgccac tagtccgagg 7140cctcgagatc cgatatcgcc
gtggcggccg ccagctgaag cttaattatc ctgggcacga 7200gtgaaacaaa
gctaaaacct ttatttagca tggccattga atgtaacaat tatatatatc
7260gcaagcacaa aaaatcaagg agagagaact accactttgt tcatgtgtac
aatgttcatt 7320atctccataa gcaaaaaaaa aaaatagaaa acatatgcta
taaggttgat attctcacga 7380gtaagcggca cttgctactt attgacattg
cagatttttg gctacagaaa tagtatatta 7440gagattataa ttgctaatca
aatcaaaata taaaattagt aaaccaaacc atttataccc 7500ttccttagta
gttatggatt gttttttaat gatatttctg caaaccaaag aaagattgtt
7560atccagatag aatttagttt tgatattcat ttttttgttg aagattgaac
gccatatctg 7620ggcctcataa ttcaaaagac ggtgccatta tcggtagcgt
ttcgcattgt actggatttc 7680agaaatttca cagttgatga atcgaaaaga
atggtctcat tgcaacacgt aaggttaaga 7740tgtccctttt taccattata
ggcaataaat gaatcataaa acgaccgtat actggtgaaa 7800tagtagggag
aacgagtacc tgtagtaaaa agtataaatc atagttaatc gggcaatgtc
7860cctcgatcaa ggagtattgt gtcatgttcg agacaaacgc caacattttt
gtttcttttg 7920gacaaatgtt gtttgcattt atgatccgtt atattttgat
ctaatgtaga gttgcacgta 7980gttcttactg gcaaagaaat cgatgcatac
caaaaaagaa taaaggtgat atttgatctt 8040taccgtttag ttccaacgta
aaattgtgcc tttggactta aaatggcgtc 8090877103DNAArtificial
SequencepSPRT190 87tcgaggcggc cgccaaaata gaatttgaag gagatgattt
gttttcctcc caagtgcctc 60cactgacgtt taaaatgttc ctgatcatca gatgaccttc
ctcggtcaag atggattccg 120gatgaaattg cacaccttct acagtgtact
tcttgtgtct tacacccatg ataattccat 180tttcggtact cgcagtaacc
ttcaagcagg atggtaggga cgattctgtc ccggccaatg 240aatggtatct
tgtcacagca ataccttgcg gcacgttctt gaaaattcca cagttgtcgt
300gagagattgg ggacgtttta ccgtggacaa tctcaccagc gtaggcaact
tcaccaccaa 360atacgtcaaa catgcattgc tggcccatac agattccaaa
tacaggaatt ttcccagtaa 420agtaccggat acagtctctt gaaatgccag
aatctgtctt tgggtggcct ggtccaggcg 480agataagcaa tgtgtcggga
ttcaaggcgg caatttctgg aactgtaatt gcatcgttac 540ggtagacgct
cactttggcg ccctcctggc acaagtactc gtaaacgttc caggtaaagg
600aatcgtagtt gtcaattaga accacatgct tattgattgg gtttgttgca
gcgtgcacag 660acatatttaa atcgaggcgc gccatgttaa ttaaatctgt
tgtttattca attttagccg 720cttctttggc caatgcaggt gcttccaagg
tgtacgaccc cgagcaacgc aaacgcatga 780tcactgggcc tcagtggtgg
gctcgctgca agcaaatgaa cgtgctggac tccttcatca 840actactatga
ttccgagaag cacgccgaga acgccgtgat ttttctgcat ggtaacgctg
900cctccagcta cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct
agatgcatca 960tccctgatct gatcggaatg ggtaagtccg gcaagagcgg
gaatggctca tatcgcctcc 1020tggatcacta caagtacctc accgcttggt
tcgagctgct gaaccttcca aagaaaatca 1080tctttgtggg ccacgactgg
ggggcttgtc tggcctttca ctactcctac gagcaccaag 1140acaagatcaa
ggccatcgtc catgctgaga gtgtcgtgga cgtgatcgag tcctgggacg
1200agtggcctga catcgaggag gatatcgccc tgatcaagag cgaagagggc
gagaaaatgg 1260tgcttgagaa taacttcttc gtcgagacca tgctcccaag
caagatcatg cggaaactgg 1320agcctgagga gttcgctgcc tacctggagc
cattcaagga gaagggcgag gttagacggc 1380ctaccctctc ctggcctcgc
gagatccctc tcgttaaggg aggcaagccc gacgtcgtcc 1440agattgtccg
caactacaac gcctaccttc gggccagcga cgatctgcct aagatgttca
1500tcgagtccga ccctgggttc ttttccaacg ctattgtcga gggagctaag
aagttcccta 1560acaccgagtt cgtgaaggtg aagggcctcc acttcagcca
ggaggacgct ccagatgaaa 1620tgggtaagta catcaagagc ttcgtggagc
gcgtgctgaa gaacgagcag taaggcgcgc 1680gggccgcatc atgtaattag
ttatgtcacg cttacattca cgccctcccc ccacatccgc 1740tctaaccgaa
aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
1800agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
ttctgtacag 1860acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
gagaaggttt tgggacgctc 1920gaaggcttta atttgcggcc ctgcattaat
gaatcggcca acgcgcgggg agaggcggtt 1980tgcgtattgg gcgctcttcc
gcttcctcgc tcactgactc gctgcgctcg gtcagatctg 2040tttagcttgc
ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
2100tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
ccgtacattt 2160agcccataca tccccatgta taatcatttg catccataca
ttttgatggc cgcacggcgc 2220gaagcaaaaa ttacggctcc tcgctgcaga
cctgcgagca gggaaacgct cccctcacag 2280acgcgttgaa ttgtccccac
gccgcgcccc tgtagagaaa tataaaaggt taggatttgc 2340cactgaggtt
cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
2400tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga
ggccgcgatt 2460aaattccaac atggatgctg atttatatgg gtataaatgg
gctcgcgata atgtcgggca 2520atcaggtgcg acaatctatc gattgtatgg
gaagcccgat gcgccagagt tgtttctgaa 2580acatggcaaa ggtagcgttg
ccaatgatgt tacagatgag atggtcagac taaactggct 2640gacggaattt
atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
2700gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag
aatatcctga 2760ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
cgccggttgc attcgattcc 2820tgtttgtaat tgtcctttta acagcgatcg
cgtatttcgt ctcgctcagg cgcaatcacg 2880aatgaataac ggtttggttg
atgcgagtga ttttgatgac gagcgtaatg gctggcctgt 2940tgaacaagtc
tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
3000tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
taggttgtat 3060tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
cttgccatcc tatggaactg 3120cctcggtgag ttttctcctt cattacagaa
acggcttttt caaaaatatg gtattgataa 3180tcctgatatg aataaattgc
agtttcattt gatgctcgat gagtttttct aatcagtact 3240gacaataaaa
agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
3300gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
cgacatcatc 3360tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
gcgtcaatcg tatgtgaatg 3420ctggtcgcta tactgctgtc gattcgatac
taacgccgcc atccagtgtc gaaaacgagc 3480tcgaattcct gggggaacaa
cttcacagaa tgttttgtca tattgtcgaa gtggtcacaa 3540aacaagagaa
gttccgccaa ttataaaaag ggaacccgta tatttcagct tcacggatga
3600tttccagggt gagagtactg tatatgggct tacgatagaa ggccataaaa
atttcttgct 3660tggcaacaaa atagaagtga aatcatgtcg aggctgctgt
gtgggagaac agcttaaaat 3720atcacaaaaa aagaatctaa aacactgtgt
tgcttgtccc agaaagggaa tcaagtattt 3780ttataaagat tggagtggta
aaaatcgagt atgtgctaga tgctatggaa gatacaaatt 3840cagcggtcat
cactgtataa attgcaagta tgtaccagaa gcacgtgaag tgaaaaaggc
3900aaaagacaaa ggcgaaaaat tgggcattac gcccgaaggt ttgccagtta
aaggaccaga 3960gtgtataaaa tgtggcggaa tcttacagtg gcctatgcgg
ccgctctaga actagtggat 4020cgatccccaa ttcgccctat agtgagtcgt
attacaattc actggccgtc gttttacaac 4080gtcgtgactg ggaaaaccct
ggcgttaccc aacttaatcg ccttgcagca catccccctt 4140tcgccagctg
gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
4200gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
tgcggtattt 4260cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
agcggcgcat taagcgcggc 4320gggtgtggtg gttacgcgca gcgtgaccgc
tacacttgcc agcgccctag cgcccgctcc 4380tttcgctttc ttcccttcct
ttctcgccac gttcgccggc tttccccgtc aagctctaaa 4440tcggggtggg
ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
4500ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
cgggctattc 4560ttttgattta taagggattt tgccgatttc ggcctattgg
ttaaaaaatg agctgattta 4620acaaaaattt aacgcgaatt ttaacaaaat
attaacgttt acaattttat ggtgcactct 4680cagtacaatc tgctctgatg
ccgcatagtt aagccagccc cgacacccgc caacacccgc 4740tgacgcgccc
tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
4800ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
cgagacgaaa 4860gggcctcgtg atacgcctat ttttataggt taatgtcatg
ataataatgg tttcttagac 4920gtcaggtggc acttttcggg gaaatgtgcg
cggaacccct atttgtttat ttttctaaat 4980acattcaaat atgtatccgc
tcatgagaca ataaccctga taaatgcttc aataatattg 5040aaaaaggaag
agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
5100attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
atgctgaaga 5160tcagttgggt gcacgagtgg gttacatcga actggatctc
aacagcggta agatccttga 5220gagttttcgc cccgaagaac gttttccaat
gatgagcact tttaaagttc tgctatgtgg 5280cgcggtatta tcccgtattg
acgccgggca agagcaactc ggtcgccgca tacactattc 5340tcagaatgac
ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
5400agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
ccaacttact 5460tctgacaacg atcggaggac cgaaggagct aaccgctttt
ttgcacaaca tgggggatca 5520tgtaactcgc cttgatcgtt gggaaccgga
gctgaatgaa gccataccaa acgacgagcg 5580tgacaccacg atgcctgtag
caatggcaac aacgttgcgc aaactattaa ctggcgaact 5640acttactcta
gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
5700accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
ctggagccgg 5760tgagcgtggg tctcgcggta tcattgcagc actggggcca
gatggtaagc cctcccgtat 5820cgtagttatc tacacgacgg ggagtcaggc
aactatggat gaacgaaata gacagatcgc 5880tgagataggt gcctcactga
ttaagcattg gtaactgtca gaccaagttt actcatatat 5940actttagatt
gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
6000tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
cgtcagaccc 6060cgtagaaaag atcaaaggat cttcttgaga tccttttttt
ctgcgcgtaa tctgctgctt 6120gcaaacaaaa aaaccaccgc taccagcggt
ggtttgtttg ccggatcaag agctaccaac 6180tctttttccg aaggtaactg
gcttcagcag agcgcagata ccaaatactg tccttctagt 6240gtagccgtag
ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
6300gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
ccgggttgga 6360ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
tgaacggggg gttcgtgcac 6420acagcccagc ttggagcgaa cgacctacac
cgaactgaga tacctacagc gtgagcattg 6480agaaagcgcc acgcttcccg
aagggagaaa ggcggacagg tatccggtaa gcggcagggt 6540cggaacagga
gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
6600tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
caggggggcg 6660gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
ttcctggcct tttgctggcc 6720ttttgctcac atgttctttc ctgcgttatc
ccctgattct gtggataacc gtattaccgc 6780ctttgagtga gctgataccg
ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 6840cgaggaagcg
gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
6900ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
gcaacgcaat 6960taatgtgagt tagctcactc attaggcacc ccaggcttta
cactttatgc ttccgcggct 7020cgtatgttgt gtggaattgt gagcggataa
caatttcaca caggaaacag ctatgaccat 7080gattacgcca ctagtccgag gcc
7103889685DNAArtificial SequencepSPRT192 88taaatctgtt gtttattcaa
ttttagccgc ttctttggcc aatgcaggtg cttccaaggt 60gtacgacccc gagcaacgca
aacgcatgat cactgggcct cagtggtggg ctcgctgcaa 120gcaaatgaac
gtgctggact ccttcatcaa ctactatgat tccgagaagc acgccgagaa
180cgccgtgatt tttctgcatg gtaacgctgc ctccagctac ctgtggaggc
acgtcgtgcc 240tcacatcgag cccgtggcta gatgcatcat ccctgatctg
atcggaatgg gtaagtccgg 300caagagcggg aatggctcat atcgcctcct
ggatcactac aagtacctca ccgcttggtt 360cgagctgctg aaccttccaa
agaaaatcat ctttgtgggc cacgactggg gggcttgtct 420ggcctttcac
tactcctacg agcaccaaga caagatcaag gccatcgtcc atgctgagag
480tgtcgtggac gtgatcgagt cctgggacga gtggcctgac atcgaggagg
atatcgccct 540gatcaagagc gaagagggcg agaaaatggt gcttgagaat
aacttcttcg tcgagaccat 600gctcccaagc aagatcatgc ggaaactgga
gcctgaggag ttcgctgcct acctggagcc 660attcaaggag aagggcgagg
ttagacggcc taccctctcc tggcctcgcg agatccctct 720cgttaaggga
ggcaagcccg acgtcgtcca gattgtccgc aactacaacg cctaccttcg
780ggccagcgac gatctgccta agatgttcat cgagtccgac cctgggttct
tttccaacgc 840tattgtcgag ggagctaaga agttccctaa caccgagttc
gtgaaggtga agggcctcca 900cttcagccag gaggacgctc cagatgaaat
gggtaagtac atcaagagct tcgtggagcg 960cgtgctgaag aacgagcagt
aaggcgcgcg ggccgcatca tgtaattagt tatgtcacgc 1020ttacattcac
gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
1080gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
gttatttata 1140tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
catgtaacat tatactgaaa 1200accttgcttg agaaggtttt gggacgctcg
aaggctttaa tttgcggccc tgcattaatg 1260aatcggccaa cgcgcgggga
gaggcggttt gcgtattggg cgctcttccg cttcctcgct 1320cactgactcg
ctgcgctcgg tcagatctcg ctgttccagt caatcagggt attgaagctc
1380atggtcttta ctccatcaca gggttccgcc ttatccggcc tacagaaccc
aaaatatcaa 1440cgcattacgt aggcctgata agcgcagcgc catcaggcgt
cagatcactc catcatcttc 1500tcgatcggca gtaccagaat cgagctggcg
ccaagcgctt tcagtttctc catggtttcc 1560cagaacaacg tttcgctgct
gaccatgtgc atcgccacgc gctgttgctc gcctgccagc 1620ggcagaattg
tcggcctttc ggcgcctggc agcagggcga taacctcttc caggcgttca
1680cttggcgcgt gcatcatgat gtatttcgat tcgcgcgcct gaatcacgcc
ctgaatacgg 1740gtcagcaatt tatcgatcag ctcttgcttg ctctgtgcca
tctcaccgtc gcgctgaatc 1800agacaggctt tagagcggta gataacttcg
acttcacgca ggccgttagc ttcaagcgtc 1860gcgccggtag agaccaaatc
gcagatagcg tcggccagcc ccgcgcgcgg cgcgacttcg 1920acagaaccat
ttaacagaca cgatttaaaa gagacgcctt tctggtcgag gtagcgtttg
1980aggaggtgcg gatatgaggt agcgatacgt ttaccgtcca gcgcggccgg
gccgtcccag 2040gcttcgtcaa ccggtgttgc cagcgataaa cggcagccgc
cgaagtcaag acggcgcagg 2100gttaaatagc gtggatcttc gccctgtgcg
cggcggttga gtagctcttc ttccagcacg 2160ttttcgccga taataccgag
atcgaccacg ccatccatta ccagacccgg aatgtcatca 2220tcacgcacgc
gcaggatatc aatcggcatg ttttccgcca tcgcaatcag gcgctgagtg
2280tgtaaattaa tttttatgcc gcagcgggcc agcaattctc gtgaatcatc
gcttaaacgg 2340cctgatttct gaatagctat gcgtaagcgg gtgttgtcta
acattctgcg ttcctcttta 2400tcctgtctga accggtctgt atcgcgcgcc
aaaaaaaaag cccccggaag atgatcttcc 2460gggggctttc tcatgcgttc
atgcaccact ggaagatcca caggacgggt gtggtcgcca 2520tgatcgcgta
gtcgatagtg gctccaagta gcgaagcgag caggactggg cggcggccaa
2580agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac
gcatatagcg 2640ctagcagcac gccatagtga ctggcgatgc tgtcggaatg
gacgatatcc cgcaagaggc 2700ccggcagtac cggcataacc aagcctatgc
ctacagcatc cagggtgacg gtgccgagga 2760tgacgatgag cgcattgtta
gatttcatac acggtgcctg actgcgttag caatttaact 2820gtgataaact
accgcattaa agctttttct ttccaatttt ttttttttcg tcattataaa
2880aatcattacg accgagattc ccgggtaata actgatataa ttaaattgaa
gctctaattt 2940gtgagtttag tatacatgca tttacttata atacagtttt
ttagttttgc tggccgcatc 3000ttctcaaata tgcttcccag cctgcttttc
tgtaacgttc accctctacc ttagcatccc 3060ttccctttgc aaatagtcct
cttccaacaa taataatgtc agatcctgta
gagaccacat 3120catccacggt tctatactgt tgacccaatg cgtctccctt
gtcatctaaa cccacaccgg 3180gtgtcataat caaccaatcg taaccttcat
ctcttccacc catgtctctt tgagcaataa 3240agccgataac aaaatctttg
tcgctcttcg caatgtcaac agtaccctta gtatattctc 3300cagtagatag
ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct
3360ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
accacaccgt 3420gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
acccgcagag tactgcaatt 3480tgactgtatt accaatgtca gcaaattttc
tgtcttcgaa gagtaaaaaa ttgtacttgg 3540cggataatgc ctttagcggc
ttaactgtgc cctccatgga aaaatcagtc aagatatcca 3600catgtgtttt
tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct
3660tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
atgatattaa 3720atagcttggc agcaacagga ctaggatgag tagcagcacg
ttccttatat gtagctttcg 3780acatgattta tcttcgtttc ctgcaggttt
ttgttctgtg cagttgggtt aagaatactg 3840ggcaatttca tgtttcttca
acactacata tgcgtatata taccaatcta agtctgtgct 3900ccttccttcg
ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc
3960gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct
tatcgatgat 4020aagctgtcaa acatgagaat taattcagat ccgctgttcc
agtcaatcag ggtattgaag 4080ctcatggtct ttactccatc acagggttcc
gccttatccg gcctacagaa cccaaaatat 4140caacgcatta cgtaggcctg
ataagcgcag cgccatcagg cgtcagatca ctccatcatc 4200ttctcgatcg
gcagtaccag aatcgagctg gcgccaagcg ctttcagttt ctccatggtt
4260tcccagaaca acgtttcgct gctgaccatg tgcatcgcca cgcgctgttg
ctcgcctgcc 4320agcggcagaa ttgtcggcct ttcggcgcct ggcagcaggg
cgataacctc ttccaggcgt 4380tcacttggcg cgtgcatcat gatgtatttc
gattcgcgcg cctgaatcac gccctgaata 4440cgggtcagca atttatcgat
cagctcttgc ttgctctgtg ccatctcacc gtcgcgctga 4500atcagacagg
ctttagagcg gtagataact tcgacttcac gcaggccgtt agcttcaagc
4560gtcgcgccgg tagagaccaa atcgcagata gcgtcggcca gccccgcgcg
cggcgcgact 4620tcgacagaac catttaacag acacgattta aaagagacgc
ctttctggtc gaggtagcgt 4680ttgaggaggt gcggatatga ggtagcgata
cgtttaccgt ccagcgcggc cgggccgtcc 4740caggcttcgt caaccggtgt
tgccagcgat aaacggcagc cgccgaagtc aagacggcgc 4800agggttaaat
agcgtggatc ttcgccctgt gcgcggcggt tgagtagctc ttcttccagc
4860acgttttcgc cgataatacc gagatcgacc acgccatcca ttaccagacc
cggaatgtca 4920tcatcacgca cgcgcaggat atcaatcggc atgttttccg
ccatcgcaat caggcgctga 4980gtgtgtaaat taatttttat gccgcagcgg
gccagcaatt ctcgtgaatc atcgcttaaa 5040cggcctgatt tctgaatagc
tatgcgtaag cgggtgttgt ctaacattct gcgttcctct 5100ttatcctgtc
tgaaccggtc tgtatcgcgc gccaaaaaaa aagcccccgg aagatgatct
5160tccgggggct ttctcatgcg ttcatgcacc actggaagat cagatctgtt
tagcttgcct 5220cgtccccgcc gggtcacccg gccagcgaca tggggtattt
aggtctacgt tgaatgaatg 5280caggtcccta ttattgacac ctacaacttt
agcaccaatt tctagagccc tttgtaattc 5340ctctttggag ttcacctcaa
cgagaggttc catgttcaaa tctttactgt agctgtacag 5400ttccttcaat
aagggttgag atagcatctt gactataaga aggacagtgt cagctccagc
5460taatcttgct tctagtattt gatacttgct gaaaataaat tcttttctca
aaacacaagg 5520cctctccttg ggaggaaatt tcaaatctag gattttcctc
acatttacta aatcctgtaa 5580cgaaccgtga aaccaatgag gttcggtcaa
tacggaaatt gcggatgcac cagcctctgc 5640gtatttgaga gcctgttcag
cagcaacagc ttttaaacaa atgggtccct tcgatggaga 5700ggcacgcttg
acttcagcaa gaacaacggc tcttttatgg gatgatgaca acaccgtgta
5760gaaatcctgt aacggtgggg caagacctaa atcatagtta gattgtaagt
cttgaaaggt 5820gaaacctggg attttagact gctcattgac gtctatttta
cgccgagcat agatacggtc 5880ggcctatgcg gccgctctag aactagtgga
tcgatcccca attcgcccta tagtgagtcg 5940tattacaatt cactggccgt
cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 6000caacttaatc
gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
6060cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
cctgatgcgg 6120tattttctcc ttacgcatct gtgcggtatt tcacaccgca
tacgtcaaag caaccatagt 6180acgcgccctg tagcggcgca ttaagcgcgg
cgggtgtggt ggttacgcgc agcgtgaccg 6240ctacacttgc cagcgcccta
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 6300cgttcgccgg
ctttccccgt caagctctaa atcggggtgg gccatcgccc tgatagacgg
6360tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
ttccaaactg 6420gaacaacact caaccctatc tcgggctatt cttttgattt
ataagggatt ttgccgattt 6480cggcctattg gttaaaaaat gagctgattt
aacaaaaatt taacgcgaat tttaacaaaa 6540tattaacgtt tacaatttta
tggtgcactc tcagtacaat ctgctctgat gccgcatagt 6600taagccagcc
ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
6660cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
cagaggtttt 6720caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
gatacgccta tttttatagg 6780ttaatgtcat gataataatg gtttcttaga
cgtcaggtgg cacttttcgg ggaaatgtgc 6840gcggaacccc tatttgttta
tttttctaaa tacattcaaa tatgtatccg ctcatgagac 6900aataaccctg
ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
6960tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
gctcacccag 7020aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
tgcacgagtg ggttacatcg 7080aactggatct caacagcggt aagatccttg
agagttttcg ccccgaagaa cgttttccaa 7140tgatgagcac ttttaaagtt
ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 7200aagagcaact
cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
7260tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
gctgccataa 7320ccatgagtga taacactgcg gccaacttac ttctgacaac
gatcggagga ccgaaggagc 7380taaccgcttt tttgcacaac atgggggatc
atgtaactcg ccttgatcgt tgggaaccgg 7440agctgaatga agccatacca
aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 7500caacgttgcg
caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
7560tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
cttccggctg 7620gctggtttat tgctgataaa tctggagccg gtgagcgtgg
gtctcgcggt atcattgcag 7680cactggggcc agatggtaag ccctcccgta
tcgtagttat ctacacgacg gggagtcagg 7740caactatgga tgaacgaaat
agacagatcg ctgagatagg tgcctcactg attaagcatt 7800ggtaactgtc
agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
7860aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
atcccttaac 7920gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
gatcaaagga tcttcttgag 7980atcctttttt tctgcgcgta atctgctgct
tgcaaacaaa aaaaccaccg ctaccagcgg 8040tggtttgttt gccggatcaa
gagctaccaa ctctttttcc gaaggtaact ggcttcagca 8100gagcgcagat
accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
8160actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
gctgctgcca 8220gtggcgataa gtcgtgtctt accgggttgg actcaagacg
atagttaccg gataaggcgc 8280agcggtcggg ctgaacgggg ggttcgtgca
cacagcccag cttggagcga acgacctaca 8340ccgaactgag atacctacag
cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa 8400aggcggacag
gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
8460cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
tgacttgagc 8520gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
gaaaaacgcc agcaacgcgg 8580cctttttacg gttcctggcc ttttgctggc
cttttgctca catgttcttt cctgcgttat 8640cccctgattc tgtggataac
cgtattaccg cctttgagtg agctgatacc gctcgccgca 8700gccgaacgac
cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
8760aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
aggtttcccg 8820actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
ttagctcact cattaggcac 8880cccaggcttt acactttatg cttccgcggc
tcgtatgttg tgtggaattg tgagcggata 8940acaatttcac acaggaaaca
gctatgacca tgattacgcc actagtccga ggcctcgagg 9000cggccgccaa
aatagaattt gaaggagatg atttgttttc ctcccaagtg cctccactga
9060cgtttaaaat gttcctgatc atcagatgac cttcctcggt caagatggat
tccggatgaa 9120attgcacacc ttctacagtg tacttcttgt gtcttacacc
catgataatt ccattttcgg 9180tactcgcagt aaccttcaag caggatggta
gggacgattc tgtcccggcc aatgaatggt 9240atcttgtcac agcaatacct
tgcggcacgt tcttgaaaat tccacagttg tcgtgagaga 9300ttggggacgt
tttaccgtgg acaatctcac cagcgtaggc aacttcacca ccaaatacgt
9360caaacatgca ttgctggccc atacagattc caaatacagg aattttccca
gtaaagtacc 9420ggatacagtc tcttgaaatg ccagaatctg tctttgggtg
gcctggtcca ggcgagataa 9480gcaatgtgtc gggattcaag gcggcaattt
ctggaactgt aattgcatcg ttacggtaga 9540cgctcacttt ggcgccctcc
tggcacaagt actcgtaaac gttccaggta aaggaatcgt 9600agttgtcaat
tagaaccaca tgcttattga ttgggtttgt tgcagcgtgc acagacatat
9660ttaaatcgag gcgcgccatg ttaat 9685
* * * * *
References