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Plasmid Files

pY-SUMOstar

Saccharomyces cerevisiae vector for regulated expression of intracellular recombinant proteins with a cleavable 6xHis-SUMO tag.

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pY-SUMOstar.dna
Map and Sequence File:    Download    Open   
Sequence Author:  LifeSensors
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BsmBI (6306) PflFI - Tth111I (6205) BspQI - SapI (6066) PciI (5949) AhdI (5061) SacII (4996) BglI (4943) NmeAIII (4914) TsoI (4663) ScaI (4580) AatII (4142) ZraI (4140) MscI * (3802) XcmI (3673) BtgZI (263) PsiI (363) NcoI - StyI (437) SbfI (473) BlpI (497) BglII (569) BsaI (758) Acc65I (776) AgeI (779) KpnI (780) BsaI (788) BamHI (794) SalI (813) EagI - NotI (826) PaeR7I - PspXI - XhoI (834) MluI (1138) BspDI - ClaI (1221) PmlI (1307) Bsu36I (1589) BstXI (1645) BsgI (1760) AarI - BfuAI - BspMI (1780) BstBI (2088) HpaI (2382) SnaBI (3024) pY-SUMOstar 6360 bp
BsmBI  (6306)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PflFI  (6205)
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  (6205)
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.
BspQI  (6066)
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  (6066)
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.
PciI  (5949)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (5061)
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.
SacII  (4996)
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.
BglI  (4943)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
NmeAIII  (4914)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
TsoI  (4663)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (4580)
1 site
A G T A C T T C A T G A
AatII  (4142)
1 site
G A C G T C C T G C A G
ZraI  (4140)
1 site
G A C G T C C T G C A G
MscI  (3802)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
XcmI  (3673)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BtgZI  (263)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
PsiI  (363)
1 site
T T A T A A A A T A T T
NcoI  (437)
1 site
C C A T G G G G T A C C
StyI  (437)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
SbfI  (473)
1 site
C C T G C A G G G G A C G T C C
BlpI  (497)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BglII  (569)
1 site
A G A T C T T C T A G A
BsaI  (758)
2 sites
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.
Acc65I  (776)
1 site
G G T A C C C C A T G G
AgeI  (779)
1 site
A C C G G T T G G C C A
KpnI  (780)
1 site
G G T A C C C C A T G G
BsaI  (788)
2 sites
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.
BamHI  (794)
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.
SalI  (813)
1 site
G T C G A C C A G C T G
EagI  (826)
1 site
C G G C C G G C C G G C
NotI  (826)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (834)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (834)
1 site
V C T C G A G B B G A G C T C V
XhoI  (834)
1 site
C T C G A G G A G C T C
MluI  (1138)
1 site
A C G C G T T G C G C A
BspDI  (1221)
1 site
A T C G A T T A G C T A
ClaI  (1221)
1 site
A T C G A T T A G C T A
PmlI  (1307)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
Bsu36I  (1589)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BstXI  (1645)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BsgI  (1760)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AarI  (1780)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (1780)
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  (1780)
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.
BstBI  (2088)
1 site
T T C G A A A A G C T T
HpaI  (2382)
1 site
G T T A A C C A A T T G
SnaBI  (3024)
1 site
T A C G T A A T G C A T
2μ ori
2383 .. 3725  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
2383 .. 3725  =  1343 bp
yeast 2μ plasmid origin of replication
AmpR
4274 .. 5134  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4274 .. 4342  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4274 .. 5134  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4343 .. 5134  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4274 .. 5134  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TRP1
1348 .. 2022  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
1348 .. 2022  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
ori
5305 .. 5893  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5305 .. 5893  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SUMO
475 .. 762  =  288 bp
96 amino acids  =  11.0 kDa
Product: cleavable ubiquitin-like protein tag
SUMO
475 .. 762  =  288 bp
96 amino acids  =  11.0 kDa
Product: cleavable ubiquitin-like protein tag
CYC1 terminator
966 .. 1213  =  248 bp
transcription terminator for CYC1
CYC1 terminator
966 .. 1213  =  248 bp
transcription terminator for CYC1
CUP1 promoter
169 .. 403  =  235 bp
inducible metallothionein promoter, regulated by Cu2+
CUP1 promoter
169 .. 403  =  235 bp
inducible metallothionein promoter, regulated by Cu2+
AmpR promoter
4169 .. 4273  =  105 bp
AmpR promoter
4169 .. 4273  =  105 bp
TRP1 promoter
1246 .. 1347  =  102 bp
TRP1 promoter
1246 .. 1347  =  102 bp
MCS
776 .. 839  =  64 bp
multiple cloning site
MCS
776 .. 839  =  64 bp
multiple cloning site
6xHis
445 .. 462  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
445 .. 462  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
439 .. 441  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
439 .. 441  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  439 .. 765  =  327 bp
ORF:  108 amino acids  =  12.4 kDa
ORF:  1348 .. 2022  =  675 bp
ORF:  224 amino acids  =  24.1 kDa
ORF:  1598 .. 1879  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  4274 .. 5134  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2418 .. 2708  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  2164 .. 2403  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  3184 .. 3504  =  321 bp
ORF:  106 amino acids  =  12.9 kDa
ORF:  4738 .. 5004  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3546 .. 3887  =  342 bp
ORF:  113 amino acids  =  13.0 kDa
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