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Plasmid Files

pYM26

Plasmid with a TRP1 marker that provides a template for PCR to fuse yeast enhanced GFP (yeGFP) to the C-terminus of a protein of interest.

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pYM26 Sequence and MappYM26.dna
Map and Sequence File   
Sequence Author:  EUROSCARF
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 PvuII (15) PfoI (4077) EcoO109I (4020) AatII (3966) ZraI (3964) SspI (3848) XmnI (3643) ScaI (3524) TsoI (3443) FspI (3266) NmeAIII (3192) BmrI (3084) BanI (2992) PspFI (2459) BseYI (2455) PciI (2151) HindIII (19) BsiWI (25) SalI (37) AarI - BamHI (43) BspEI * (46) BstXI (248) MscI (252) MfeI (641) PflMI (777) BsrGI (788) BssHII (801) SnaBI (851) StuI (896) BseRI (942) XbaI (1274) BstAPI (1475) BbsI (1845) AbsI - PaeR7I - PspXI - XhoI (1862) Eco53kI (1872) BanII - SacI (1874) EcoRI (1876) BspDI - ClaI (1883) EcoRV (1890) SfiI (1913) SacII (1920) BspQI - SapI (2035) pYM26 4298 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
PfoI  (4077)
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.
EcoO109I  (4020)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (3966)
1 site
G A C G T C C T G C A G
ZraI  (3964)
1 site
G A C G T C C T G C A G
SspI  (3848)
1 site
A A T A T T T T A T A A
XmnI  (3643)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3524)
1 site
A G T A C T T C A T G A
TsoI  (3443)
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).
FspI  (3266)
1 site
T G C G C A A C G C G T
NmeAIII  (3192)
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).
BmrI  (3084)
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.
BanI  (2992)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
PspFI  (2459)
1 site
C C C A G C G G G T C G
BseYI  (2455)
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.
PciI  (2151)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
HindIII  (19)
1 site
A A G C T T T T C G A A
BsiWI  (25)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
SalI  (37)
1 site
G T C G A C C A G C T G
AarI  (43)
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.
BamHI  (43)
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.
BspEI  (46)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
BstXI  (248)
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.
MscI  (252)
1 site
T G G C C A A C C G G T
MfeI  (641)
1 site
C A A T T G G T T A A C
PflMI  (777)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BsrGI  (788)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BssHII  (801)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
SnaBI  (851)
1 site
T A C G T A A T G C A T
StuI  (896)
1 site
A G G C C T T C C G G A
BseRI  (942)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
XbaI  (1274)
1 site
T C T A G A A G A T C T
BstAPI  (1475)
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.
BbsI  (1845)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AbsI  (1862)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (1862)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1862)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1862)
1 site
C T C G A G G A G C T C
Eco53kI  (1872)
1 site
G A G C T C C T C G A G
BanII  (1874)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (1874)
1 site
G A G C T C C T C G A G
EcoRI  (1876)
1 site
G A A T T C C T T A A G
BspDI  (1883)
1 site
A T C G A T T A G C T A
ClaI  (1883)
1 site
A T C G A T T A G C T A
EcoRV  (1890)
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.
SfiI  (1913)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SacII  (1920)
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.
BspQI  (2035)
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  (2035)
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.
AmpR
2971 .. 3831  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2971 .. 3762  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2971 .. 3831  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3763 .. 3831  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2971 .. 3831  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
yEGFP
85 .. 798  =  714 bp
237 amino acids  =  26.7 kDa
Product: yeast-enhanced green fluorescent protein
codons optimized for translation in S. cerevisiae and
C. albicans
yEGFP
85 .. 798  =  714 bp
237 amino acids  =  26.7 kDa
Product: yeast-enhanced green fluorescent protein
codons optimized for translation in S. cerevisiae and
C. albicans
KlTRP1
886 .. 1518  =  633 bp
210 amino acids  =  22.9 kDa
Product: phosphoribosylanthranilate isomerase,
required for tryptophan biosynthesis
K. lactis auxotrophic marker that also works in S.
cerevisiae
KlTRP1
886 .. 1518  =  633 bp
210 amino acids  =  22.9 kDa
Product: phosphoribosylanthranilate isomerase,
required for tryptophan biosynthesis
K. lactis auxotrophic marker that also works in S.
cerevisiae
ori
2212 .. 2800  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2212 .. 2800  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
3832 .. 3936  =  105 bp
AmpR promoter
3832 .. 3936  =  105 bp
SP6 promoter
4282 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
4282 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
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