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Plasmid Files

pRS424

Yeast episomal vector with a TRP1 marker and an MCS derived from pBLUESCRIPT II.

 
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pRS424.dna
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XcmI (5496) BseRI (5483) BmgBI (5285) BbsI (5127) NsiI (4495) ScaI (3767) NmeAIII (3435) BsaI (3348) AhdI (3287) AlwNI (2810) PfoI (46) BspEI * (332) PmlI (427) BstZ17I (434) BstAPI (546) Bsu36I (709) BsgI (880) AarI - BfuAI - BspMI (900) PsiI (1337) DraIII (1465) BtgZI (1466) NgoMIV (1566) NaeI (1568) Acc65I (1894) KpnI (1898) PspOMI (1900) ApaI (1904) AbsI - PaeR7I - PspXI - XhoI (1909) SalI (1915) HincII (1917) BspDI - ClaI (1925) EcoRI (1942) PstI (1952) TspMI - XmaI (1954) SmaI (1956) BamHI (1960) SpeI (1966) EagI - NotI (1979) BtgI (1988) AleI (1990) SacII (1991) Eco53kI (1998) SacI (2000) BspQI - SapI (2278) AflIII - PciI (2394) pRS424 5616 bp
XcmI  (5496)
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.
BseRI  (5483)
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.
BmgBI  (5285)
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.
BbsI  (5127)
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.
NsiI  (4495)
1 site
A T G C A T T A C G T A
ScaI  (3767)
1 site
A G T A C T T C A T G A
NmeAIII  (3435)
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).
BsaI  (3348)
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  (3287)
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  (2810)
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.
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.
BspEI  (332)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PmlI  (427)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstZ17I  (434)
1 site
G T A T A C C A T A T G
BstAPI  (546)
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.
Bsu36I  (709)
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.
BsgI  (880)
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  (900)
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  (900)
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  (900)
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.
PsiI  (1337)
1 site
T T A T A A A A T A T T
DraIII  (1465)
1 site
C A C N N N G T G G T G N N N C A C