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Plasmid Files

pYM25

Plasmid with a hygromycin resistance 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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pYM25 Sequence and MappYM25.dna
Map and Sequence File   
Sequence Author:  EUROSCARF
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 HindIII (19) PvuII (15) NotI (6) PfoI (4684) SspI (4455) XmnI (4250) BmrI (3691) PciI (2758) BspQI - SapI (2642) NotI (2521) SfiI (2520) SpeI (2507) EcoRV (2497) BspDI - ClaI (2490) BanII - SacI (2481) BsiWI (25) SalI (37) AccI (38) AarI - BamHI (43) MscI (252) MfeI (641) PflMI (777) BssHII (801) BglII (808) BstEII (838) BmgBI (891) Bpu10I (900) BseRI (1002) PshAI (1227) AsiSI (1564) RsrII (1608) BtgZI (1872) AvaI - BsoBI - PaeR7I - XhoI (2227) BmeT110I (2228) PpuMI (2324) Eco53kI (2479) pYM25 4905 bp
HindIII  (19)
1 site
A A G C T T T T C G A A
PvuII  (15)
1 site
C A G C T G G T C G A C
NotI  (6)
2 sites
G C G G C C G C C G C C G G C G
PfoI  (4684)
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.
SspI  (4455)
1 site
A A T A T T T T A T A A
XmnI  (4250)
1 site
G A A N N N N T T C C T T N N N N A A G
BmrI  (3691)
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.
PciI  (2758)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2642)
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  (2642)
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.
NotI  (2521)
2 sites
G C G G C C G C C G C C G G C G
SfiI  (2520)
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.
SpeI  (2507)
1 site
A C T A G T T G A T C A
EcoRV  (2497)
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.
BspDI  (2490)
1 site
A T C G A T T A G C T A
ClaI  (2490)
1 site
A T C G A T T A G C T A
BanII  (2481)
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  (2481)
1 site
G A G C T C C T C G A G
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
AccI  (38)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
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.
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.
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.
BglII  (808)
1 site
A G A T C T T C T A G A
BstEII  (838)
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.
BmgBI  (891)
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.
Bpu10I  (900)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BseRI  (1002)
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.
PshAI  (1227)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AsiSI  (1564)
1 site
G C G A T C G C C G C T A G C G
RsrII  (1608)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BtgZI  (1872)
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.
AvaI  (2227)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2227)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PaeR7I  (2227)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2227)
1 site
C T C G A G G A G C T C
BmeT110I  (2228)
1 site
C Y C G R G G R G C Y C
PpuMI  (2324)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
Eco53kI  (2479)
1 site
G A G C T C C T C G A G
HygR
1197 .. 2225  =  1029 bp
342 amino acids  =  38.1 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
1197 .. 2225  =  1029 bp
342 amino acids  =  38.1 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
3578 .. 4438  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3578 .. 4369  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3578 .. 4438  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4370 .. 4438  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3578 .. 4438  =  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
ori
2819 .. 3407  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2819 .. 3407  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
TEF promoter
853 .. 1196  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
853 .. 1196  =  344 bp
Ashbya gossypii TEF promoter
CYC1 terminator
2279 .. 2468  =  190 bp
transcription terminator for CYC1
CYC1 terminator
2279 .. 2468  =  190 bp
transcription terminator for CYC1
AmpR promoter
4439 .. 4543  =  105 bp
AmpR promoter
4439 .. 4543  =  105 bp
SP6 promoter
4889 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
4889 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
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