pAS1 HA epitope and polylinker reading frame
R1 NheI
GAA TTC ATG GCT
TAC CCA TAC GAT GTT CCA GAT TAC GCT AGC TTG GGT GGT
SfiI .
CAT ATG GCC ATG
GAG GCC CCG GGG ATC C
NdeI NcoI SmaI
BamHI
pACT2 HA epitope
and polylinker
NheI
ATG GCT TAC CCA
TAC GAT GTT CCA GAT TAC GCT AGC TTG GGT GGT
SfiI .
CAT ATG GCC ATG
GAG GCC CCG GGG ATC CGA ATT CGA GCT CGA GAG ATC TAT
NdeI NcoI SmaI
BamHI R1 SacI XhoI BglII
GAATCGTAGATACTGAAAAACCCCGCAAGTTCACTTCAACTGTGCATCGTGCACCATCTC
AATTTCTTTCATTTATACATCGTTTTGCCT
Sequence around
the cloning site of pACT (pSE1107)
Mlu I
GCGTATAACGCGTTTGGAATCACTACAGGGATGTTTAATACCACTACAATGGATGATGT
ATAT AAC TAT CTA
TTC GAT GAT GAA GAT ACC CCA CCA AAC CCA AAA AAA
Bgl II BamH Bgl
II
GAG ATC TGG AAT
TCG GAT CCT CGA GAG ATC TAT
EcoRI XhoI
GAATCGTAGATACTGAAAAACCCCGCAAGTTCACTTCAACTGTGCATCGTGCACCATCTC
AATTTCTTTCATTTATACATCGTTTTGCCT
Sequencing primers
pACT forward 5'
5' CTATCTATTCGATGATGAAG
pACT Reverse 3'
5' ACAGTTGAAGTGAACTTGCG
GAL Promoter Forward
5' TACTTTAACGTCAAGGAGAA
lacZ 5' agctggcgtaatagcgaagag
(Reverse primer
for Lambda ADH and YES)
ADH Forward 5'
TCTGCACAATATTTCAAGCT
The sequences of
the PCR primers we have used successfully are:
1)
TAA TAC GAC TCA
CTA TAG GGA GAC CAC ATG GAT GAT GTA TAT AAC TAT CTA TTC
T7 Promoter Met
Gal4 Activation Domain
21 AA before the
BglII site
2)
CTA CCA GAA TTC
GGC ATG CCG GTA GAG GTG TGG TCA
In the ADH Terminator
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