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pRS423

Yeast episomal vector with a HIS3 marker and an MCS derived from pBLUESCRIPT II.

 
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pRS423.dna
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MfeI (5679) XcmI (5678) BmgBI (5467) SnaBI (5029) BsaHI (4006) ScaI (3949) NmeAIII (3617) BsaI (3530) AhdI (3469) AlwNI (2992) PspFI (2884) PfoI (46) BsaBI * (561) NdeI (694) MscI (711) BsmI (716) BsiWI (916) NheI (1008) BmtI (1012) BclI * (1052) PsiI (1519) DraIII (1647) BtgZI (1648) NgoMIV (1748) NaeI (1750) PspOMI (2082) ApaI (2086) AbsI - PaeR7I - PspXI - XhoI (2091) SalI (2097) HincII (2099) BspDI - ClaI (2107) EcoRV (2120) EcoRI (2124) TspMI - XmaI (2136) SmaI (2138) BamHI (2142) SpeI (2148) EagI - NotI (2161) BtgI (2170) SacII (2173) Eco53kI (2180) SacI (2182) BspQI - SapI (2460) BseYI (2880) pRS423 5797 bp
MfeI  (5679)
1 site
C A A T T G G T T A A C
XcmI  (5678)
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.
BmgBI  (5467)
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  (5029)
1 site
T A C G T A A T G C A T
BsaHI  (4006)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
ScaI  (3949)
1 site
A G T A C T T C A T G A
NmeAIII  (3617)
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  (3530)
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  (3469)
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  (2992)
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  (2884)
1 site
C C C A G C G G G T C G
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.
BsaBI  (561)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
NdeI  (694)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
MscI  (711)
1 site
T G G C C A A C C G G T
BsmI  (716)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BsiWI  (916)
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.
NheI  (1008)
1 site
G C T A G C C G A T C G
BmtI  (1012)
1 site
G C T A G C C G A T C G
BclI  (1052)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PsiI  (1519)
1 site
T T A T A A A A T A T T
DraIII  (1647)
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  (1648)
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  (1748)
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  (1750)
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  (2082)
1 site
G G G C C C C C C G G G
ApaI  (2086)
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  (2091)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2091)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2091)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2091)
1 site
C T C G A G G A G C T C
SalI  (2097)
1 site
G T C G A C C A G C T G
HincII  (2099)
1 site
G T Y R A C C A R Y T G
BspDI  (2107)
1 site
A T C G A T T A G C T A
ClaI  (2107)
1 site
A T