pBluescript SK(+)

Standard cloning vector (phagemid excised from lambda ZAP). The f1 (+) orientation allows rescue of sense strand ssDNA. pBluescript KS(+) has a reversed MCS.

Sequence Author: Stratagene

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Basic Cloning Vectors | More Plasmid Sets
No matches
XmnI (2645) BsaHI (2583) ScaI (2526) TatI (2524) NmeAIII (2194) BsaI (2107) AhdI (2046) AlwNI (1569) PspFI (1461) PsiI (102) BsaAI - DraIII (230) BtgZI (231) NgoMIV (331) NaeI (333) 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) pBluescript SK(+) 2958 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  (102)
1 site
T T A T A A A A T A T T
BsaAI  (230)
1 site
Y A C G T R R T G C A Y
DraIII  (230)
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  (231)
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  (331)
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  (333)
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
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
Eco53kI  (757)
1 site
G A G C T C C T C G A G
SacI  (759)
1 site
G A G C T C C T C G A G
BspQI  (1037)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1037)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
AflIII  (1153)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (1153)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (1157)
1 site
R C A T G Y Y G T A C R
BseYI  (1457)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AmpR
1973 .. 2833  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1973 .. 2764  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1973 .. 2833  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2765 .. 2833  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1973 .. 2833  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1214 .. 1802  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1214 .. 1802  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
244 .. 816  =  573 bp
190 amino acids  =  20.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
244 .. 816  =  573 bp
190 amino acids  =  20.4 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
2834 .. 2938  =  105 bp
AmpR promoter
2834 .. 2938  =  105 bp
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 3:  -10  
   860 .. 866  =  7 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 2:  
   867 .. 884  =  18 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 1:  -35  
   885 .. 890  =  6 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
836 .. 852  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
836 .. 852  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
f1 ori
6 .. 461  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6 .. 461  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
MCS
653 .. 760  =  108 bp
pBluescript multiple cloning site
MCS
653 .. 760  =  108 bp
pBluescript multiple cloning site
T7 promoter
626 .. 644  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
626 .. 644  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
773 .. 791  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
773 .. 791  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
603 .. 619  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
603 .. 619  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
812 .. 828  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
812 .. 828  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2103 .. 2369  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  244 .. 816  =  573 bp
ORF:  190 amino acids  =  20.4 kDa
ORF:  1973 .. 2833  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
Click here to try SnapGene

Download pBluescript SK(+).dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps