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pBS SK(-)

Phagemid cloning vector, also known as BlueScribe SK Minus or BlueSKm. The MCS is reversed relative to pBS KS(–).

 
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pBS SK(-).dna
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NgoMIV (128) XmnI (2645) BsaHI (2583) ScaI (2526) TatI (2524) NmeAIII (2194) BsaI (2107) AhdI (2046) AlwNI (1569) PspFI (1461) NaeI (130) BtgZI (228) BsaAI (233) DraIII (236) PsiI (361) Acc65I (653) KpnI (657) PspOMI (659) EcoO109I (660) ApaI (663) AbsI - PaeR7I - PspXI - XhoI (668) SalI (674) AccI (675) HincII (676) BspDI - ClaI (684) HindIII (689) EcoRV (697) EcoRI (701) PstI (711) TspMI - XmaI (713) SmaI (715) BamHI (719) SpeI (725) XbaI (731) EagI - NotI (738) BtgI (747) AleI (749) SacII (750) BstXI (751) Eco53kI (757) SacI (759) BspQI - SapI (1037) AflIII - PciI (1153) NspI (1157) BseYI (1457) pBS SK(-) 2964 bp
NgoMIV  (128)
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.
XmnI  (2645)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (2583)
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  (2526)
1 site
A G T A C T T C A T G A
TatI  (2524)
1 site
W G T A C W W C A T G W
NmeAIII  (2194)
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  (2107)
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  (2046)
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  (1569)
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  (1461)
1 site
C C C A G C G G G T C G
NaeI  (130)
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.
BtgZI  (228)
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.
BsaAI  (233)
1 site
Y A C G T R R T G C A Y
DraIII  (236)
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.
PsiI  (361)
1 site
T T A T A A A A T A T T
Acc65I  (653)
1 site
G G T A C C C C A T G G
KpnI  (657)
1 site
G G T A C C C C A T G G
PspOMI  (659)
1 site
G G G C C C C C C G G G
EcoO109I  (660)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
ApaI  (663)
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  (668)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (668)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (668)
1 site
V C T C G A G B B G A G C T C V
XhoI  (668)
1 site
C T C G A G G A G C T C
SalI  (674)
1 site
G T C G A C C A G C T G
AccI  (675)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (676)
1 site
G T Y R A C C A R Y T G
BspDI  (684)
1 site
A T C G A T T A G C T A
ClaI  (684)
1 site
A T C G A T T A G C T A
HindIII  (689)
1 site
A A G C T T T T C G A A
EcoRV  (697)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
EcoRI  (701)
1 site
G A A T T C C T T A A G
PstI  (711)
1 site
C T G C A G G A C G T C
TspMI  (713)
1 site
C C C G G G G G G C C C
XmaI  (713)
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  (715)
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  (719)
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  (725)
1 site
A C T A G T T G A T C A
XbaI  (731)
1 site
T C T A G A A G A T C T
EagI  (738)
1 site
C G G C C G G C C G G C
NotI  (738)
1 site
G C G G C C G C C G C C G G C G
BtgI  (747)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
AleI  (749)
1 site
C A C N N N N G T G G T G N N N N C A C
SacII  (750)
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  (751)
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.