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Plasmid Files

pRS404

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

 
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pRS404.dna
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AatII (4209) ZraI (4207) ScaI (3767) NmeAIII (3435) BsaI (3348) AhdI (3287) AlwNI (2810) AflIII - PciI (2394) BspQI - SapI (2278) PfoI (46) SnaBI (319) BspEI * (332) PmlI (427) BstZ17I (434) BstAPI (546) MfeI (607) 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) pRS404 4274 bp
AatII  (4209)
1 site
G A C G T C C T G C A G
ZraI  (4207)
1 site
G A C G T C C T G C A G
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.
AflIII  (2394)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2394)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2278)
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  (2278)
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.
SnaBI  (319)
1 site
T A C G T A A T G C A T
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.
MfeI  (607)
1 site
C A A T T G G T T A A C
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

Sticky ends from different DraIII sites may not be compatible.
BtgZI  (1466)
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  (1566)
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  (1568)
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.
Acc65I  (1894)
1 site
G G T A C C C C A