Resources
Plasmid Files

pRS316

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

 
To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pRS316.dna
Map and Sequence File:    Download    Open   
Download Free Trial Get SnapGene Viewer


SwaI (4343) PmlI (4319) BsaHI (3923) AhdI (3386) PfoI (46) NdeI (324) BsgI (357) SbfI (393) BfuAI - BspMI (396) XcmI (539) NcoI (616) BstBI (676) BsmI (773) StuI (847) Bpu10I (1141) NsiI (1238) BtgZI (1568) NgoMIV (1668) NaeI (1670) Eco53kI (1995) SacI (1997) AleI (2003) SacII (2004) BstXI (2005) EagI - NotI (2010) XbaI (2017) SpeI (2023) BamHI (2029) TspMI - XmaI (2035) SmaI (2037) EcoRI (2047) HindIII (2059) BspDI - ClaI (2066) SalI (2074) AbsI - PaeR7I - PspXI - XhoI (2080) Acc65I (2095) KpnI (2099) pRS316 4887 bp
SwaI  (4343)
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  (4319)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BsaHI  (3923)
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.
AhdI  (3386)
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  (324)
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  (357)
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  (393)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (396)
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  (396)
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  (539)
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  (616)
1 site
C C A T G G G G T A C C
BstBI  (676)
1 site
T T C G A A A A G C T T
BsmI  (773)
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  (847)
1 site
A G G C C T T C C G G A
Bpu10I  (1141)
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  (1238)
1 site
A T G C A T T A C G T A
BtgZI  (1568)
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  (1668)
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  (1670)
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.
Eco53kI  (1995)
1 site
G A G C T C C T C G A G
SacI  (1997)
1 site
G A G C T C C T C G A G
AleI  (2003)
1 site
C A C N N N N G T G G T G N N N N C A C
SacII  (2004)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstXI  (2005)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
EagI  (2010)
1 site
C G G C C G G C C G G C
NotI  (2010)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2017)
1 site
T C T A G A A G A T C T
SpeI  (2023)
1 site
A C T A G T T G A T C A
BamHI  (2029)
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.
TspMI  (2035)
1 site
C C C G G G G G G C C C
XmaI  (2035)
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  (2037)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (2047)
1 site
G A A T T C C T T A A G
HindIII  (2059)
1 site
A A G C T T T T C G A A
BspDI  (2066)
1 site
A T C G A T T A G C T A
ClaI  (2066)
1 site
A T C G A T T A G C T A
SalI  (2074)
1 site
G T C G A C C A G C T G
AbsI  (2080)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2080)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2080)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2080)
1 site
C T C G A G G A G C T C
Acc65I  (2095)
1 site
G