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Plasmid Files

pRS425

Yeast episomal vector with a LEU2 marker and an MCS derived from pBLUESCRIPT II.

 
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pRS425.dna
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BmgBI (6518) SnaBI (6080) NsiI (5728) ScaI (5000) BsaI (4581) AhdI (4520) AlwNI (4043) PspFI (3935) BseYI (3931) BspQI - SapI (3511) PfoI (46) PflFI - Tth111I (192) BsrGI (632) BfuAI - BspMI (759) AflII (1438) AgeI (1503) BstEII (1718) PpuMI (1875) HpaI (2174) KasI (2203) NarI (2204) SfoI (2205) PluTI (2207) DraIII (2698) BtgZI (2699) NgoMIV (2799) NaeI (2801) PspOMI (3133) ApaI (3137) AbsI - PaeR7I - PspXI - XhoI (3142) SalI (3148) HindIII (3163) PstI (3185) TspMI - XmaI (3187) SmaI (3189) BamHI (3193) SpeI (3199) EagI - NotI (3212) AleI (3223) SacII (3224) Eco53kI (3231) SacI (3233) pRS425 6849 bp
BmgBI  (6518)
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.
SnaBI  (6080)
1 site
T A C G T A A T G C A T
NsiI  (5728)
1 site
A T G C A T T A C G T A
ScaI  (5000)
1 site
A G T A C T T C A T G A
BsaI  (4581)
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.
AhdI  (4520)
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  (4043)
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  (3935)
1 site
C C C A G C G G G T C G
BseYI  (3931)
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  (3511)
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  (3511)
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.
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.
BsrGI  (632)
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.
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.
AflII  (1438)
1 site
C T T A A G G A A T T C
AgeI  (1503)
1 site
A C C G G T T G G C C A
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.
PpuMI  (1875)
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.
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.
BtgZI  (2699)
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.
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.
PspOMI  (3133)
1 site
G G G C C C C C C G G G
ApaI  (3137)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AbsI  (3142)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (3142)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3142)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3142)
1 site
C T C G A G G A G C T C
SalI  (3148)
1 site
G T C G A C C A G C T G
HindIII  (3163)
1 site
A