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Plasmid Files

pRS416

Yeast centromere vector with a URA3 marker and an MCS derived from pBLUESCRIPT II.

 
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pRS416.dna
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SwaI (4354) PmlI (4330) BsaHI (3934) NmeAIII (3545) AhdI (3397) PfoI (46) NdeI (330) BsgI (363) SbfI (399) BfuAI - BspMI (402) XcmI (545) NcoI (622) BstBI (682) BsmI (779) StuI (853) Bpu10I (1147) NsiI (1244) BtgZI (1576) NgoMIV (1676) NaeI (1678) Acc65I (2004) KpnI (2008) AbsI - PaeR7I - PspXI - XhoI (2019) SalI (2025) BspDI - ClaI (2035) HindIII (2040) EcoRI (2052) TspMI - XmaI (2064) SmaI (2066) BamHI (2070) SpeI (2076) XbaI (2082) EagI - NotI (2089) AleI (2100) SacII (2101) BstXI (2102) Eco53kI (2108) SacI (2110) pRS416 4898 bp
SwaI  (4354)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
PmlI  (4330)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BsaHI  (3934)
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.
NmeAIII  (3545)
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).
AhdI  (3397)
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.
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.
NdeI  (330)
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.
BsgI  (363)
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).
SbfI  (399)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (402)
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  (402)
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.
XcmI  (545)
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.
NcoI  (622)
1 site
C C A T G G G G T A C C
BstBI  (682)
1 site
T T C G A A A A G C T T
BsmI  (779)
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.
StuI  (853)
1 site
A G G C C T T C C G G A
Bpu10I  (1147)
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.
NsiI  (1244)
1 site
A T G C A T T A C G T A
BtgZI  (1576)
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  (1676)
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  (1678)
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  (2004)
1 site
G G T A C C C C A T G G
KpnI  (2008)
1 site
G G T A C C C C A T G G
AbsI  (2019)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2019)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2019)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2019)
1 site
C T C G A G G A G C T C
SalI  (2025)
1 site
G T C G A C C A G C T G
BspDI  (2035)
1 site
A T C G A T T A G C T A
ClaI  (2035)
1 site
A T C G A T T A G C T A
HindIII  (2040)
1 site
A A G C T T T T C G A A
EcoRI  (2052)
1 site
G A A T T C C T T A A G
TspMI  (2064)
1 site
C C C G G G G G G C C C
XmaI  (2064)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (2066)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (2070)
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.
SpeI  (2076)
1 site
A C T A G T T G A T C A
XbaI  (2082)
1 site
T C T A G A A G A T C T
EagI  (2089)
1 site
C G G C C G G C C G G C
NotI  (2089)
1 site
G C G G C C G C C G C C G G C G
AleI  (2100)
1 site
C