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pPCR-Script Amp SK(+)

Derivative of pBluescript II SK(+) with an SrfI site for inserting blunt PCR products.

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pPCR-Script Amp SK(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Agilent Technologies
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XmnI (2645) BsaHI (2583) ScaI (2526) TatI (2524) NmeAIII (2194) BsaI (2107) AhdI (2046) AlwNI (1569) PspFI (1461) PsiI (99) BsaAI - DraIII (227) BtgZI (228) NgoMIV (328) NaeI (330) 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) BamHI (719) SrfI (728) 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) pPCR-Script Amp SK(+) 2961 bp
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
PsiI  (99)
1 site
T T A T A A A A T A T T
BsaAI  (227)
1 site
Y A C G T R R T G C A Y
DraIII  (227)
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  (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.
NgoMIV  (328)
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  (330)
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  (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
BamHI  (719)
1 site
G G A T C