pESC-LEU

Yeast episomal vector with a LEU2 marker, for galactose-regulated expression and tagging of up to two genes.

Sequence Author: Agilent Technologies

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BmgBI (7427) XbaI (7326) SnaBI (6989) NsiI (6637) ScaI (5909) BsaI (5490) BmrI (5469) AhdI (5429) AlwNI (4952) PspFI (4844) BseYI (4840) BspQI - SapI (4420) MluI (4291) BmtI (4159) NheI (4155) SacII (4153) HindIII (4140) PaeR7I - PspXI - XhoI (4133) SalI (4097) SmaI - SrfI (4092) ApaI (4091) TspMI - XmaI (4090) PspOMI (4087) BamHI (4063) BstAPI (3932) PfoI (46) PflFI - Tth111I (192) BfuAI - BspMI (759) EcoRV (1007) AflII (1438) BstEII (1718) HpaI (2174) KasI (2203) NarI (2204) SfoI (2205) PluTI (2207) DraIII (2698) NgoMIV (2799) NaeI (2801) BstZ17I (3117) PacI (3311) Eco53kI (3317) SacI (3319) BglII (3321) SpeI (3356) EagI - NotI (3363) pESC-LEU 7758 bp
BmgBI  (7427)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
XbaI  (7326)
1 site
T C T A G A A G A T C T
SnaBI  (6989)
1 site
T A C G T A A T G C A T
NsiI  (6637)
1 site
A T G C A T T A C G T A
ScaI  (5909)
1 site
A G T A C T T C A T G A
BsaI  (5490)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BmrI  (5469)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
AhdI  (5429)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (4952)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PspFI  (4844)
1 site
C C C A G C G G G T C G
BseYI  (4840)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BspQI  (4420)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (4420)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
MluI  (4291)
1 site
A C G C G T T G C G C A
BmtI  (4159)
1 site
G C T A G C C G A T C G
NheI  (4155)
1 site
G C T A G C C G A T C G
SacII  (4153)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
HindIII  (4140)
1 site
A A G C T T T T C G A A
PaeR7I  (4133)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4133)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4133)
1 site
C T C G A G G A G C T C
SalI  (4097)
1 site
G T C G A C C A G C T G
SmaI  (4092)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (4092)
1 site
G C C C G G G C C G G G C C C G
ApaI  (4091)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
TspMI  (4090)
1 site
C C C G G G G G G C C C
XmaI  (4090)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
PspOMI  (4087)
1 site
G G G C C C C C C G G G
BamHI  (4063)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BstAPI  (3932)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PfoI  (46)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PflFI  (192)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (192)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BfuAI  (759)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (759)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
EcoRV  (1007)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
AflII  (1438)
1 site
C T T A A G G A A T T C
BstEII  (1718)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
HpaI  (2174)
1 site
G T T A A C C A A T T G
KasI  (2203)
1 site
G G C G C C C C G C G G
NarI  (2204)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2205)
1 site
G G C G C C C C G C G G
PluTI  (2207)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
DraIII  (2698)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
NgoMIV  (2799)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (2801)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
BstZ17I  (3117)
1 site
G T A T A C C A T A T G
PacI  (3311)
1 site
T T A A T T A A A A T T A A T T
Eco53kI  (3317)
1 site
G A G C T C C T C G A G
SacI  (3319)
1 site
G A G C T C C T C G A G
BglII  (3321)
1 site
A G A T C T T C T A G A
SpeI  (3356)
1 site
A C T A G T T G A T C A
EagI  (3363)
1 site
C G G C C G G C C G G C
NotI  (3363)
1 site
G C G G C C G C C G C C G G C G
2μ ori
6348 .. 7690  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
6348 .. 7690  =  1343 bp
yeast 2μ plasmid origin of replication
LEU2
663 .. 1757  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
663 .. 1757  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
AmpR
5356 .. 6216  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5356 .. 6147  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5356 .. 6216  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6148 .. 6216  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5356 .. 6216  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
GAL1,10 promoter
3395 .. 4059  =  665 bp
divergent inducible promoter, regulated by Gal4
GAL1,10 promoter
3395 .. 4059  =  665 bp
divergent inducible promoter, regulated by Gal4
ori
4597 .. 5185  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4597 .. 5185  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2474 .. 2929  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2474 .. 2929  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
LEU2 promoter
1770 .. 2174  =  405 bp
LEU2 promoter
1770 .. 2174  =  405 bp
CYC1 terminator
4165 .. 4354  =  190 bp
transcription terminator for CYC1
CYC1 terminator
4165 .. 4354  =  190 bp
transcription terminator for CYC1
ADH1 terminator
3012 .. 3177  =  166 bp
transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene
ADH1 terminator
3012 .. 3177  =  166 bp
transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene
AmpR promoter
6217 .. 6321  =  105 bp
AmpR promoter
6217 .. 6321  =  105 bp
MCS 2
4063 .. 4160  =  98 bp
multiple cloning site 2
MCS 2
4063 .. 4160  =  98 bp
multiple cloning site 2
MCS 1
3307 .. 3369  =  63 bp
multiple cloning site 1
MCS 1
3307 .. 3369  =  63 bp
multiple cloning site 1
UAS
3611 .. 3728  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
UAS
3611 .. 3728  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
Myc
4106 .. 4135  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
4106 .. 4135  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
3325 .. 3348  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
3325 .. 3348  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
ORF:  784 .. 1095  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  5486 .. 5752  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6383 .. 6673  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  5356 .. 6216  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  663 .. 1769  =  1107 bp
ORF:  368 amino acids  =  39.4 kDa
ORF:  2712 .. 3023  =  312 bp
ORF:  103 amino acids  =  11.3 kDa
ORF:  7149 .. 7469  =  321 bp
ORF:  106 amino acids  =  12.9 kDa
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