pBSK(+) Simple-Kan

Subcloning vector derived from pBluescript II SK(+), with most of the restriction sites removed from the MCS and with a kanamycin resistance marker. See also pBSK(+) Simple-Amp.

Sequence Author: Biomatik

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HindIII (3035) SfiI (3029) EagI (3021) BmrI (2944) FspI (2843) NaeI (2694) NgoMIV (2692) BtgZI (2592) DraIII (2591) PsiI (2463) BsrDI (2209) NruI (2039) PasI (1821) EcoNI (1783) AsiSI (1698) Bpu10I - BsmBI - Esp3I (1676) AvaI - BmeT110I - BsoBI - TspMI - XmaI (1) SmaI (3) EcoRV (8) BpmI - PmlI (13) HindIII (17) T3 promoter lac operator BspQI - SapI (307) AflIII - PciI (423) NspI (427) BssSαI (596) BseYI (727) PspFI (731) ApaLI (737) BaeGI - Bme1580I - BsiHKAI (741) AlwNI (839) AcuI (971) PflMI (1436) pBSK(+) Simple-Kan 3040 bp
HindIII  (3035)
2 sites
A A G C T T T T C G A A
SfiI  (3029)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
EagI  (3021)
1 site
C G G C C G G C C G G C
BmrI  (2944)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
FspI  (2843)
1 site
T G C G C A A C G C G T
NaeI  (2694)
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.
NgoMIV  (2692)
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.
BtgZI  (2592)
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.
DraIII  (2591)
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  (2463)
1 site
T T A T A A A A T A T T
BsrDI  (2209)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
NruI  (2039)
1 site
T C G C G A A G C G C T
PasI  (1821)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoNI  (1783)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
AsiSI  (1698)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (1676)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BsmBI  (1676)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (1676)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
AvaI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BmeT110I  (1)
1 site
C Y C G R G G R G C Y C
BsoBI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (1)
1 site
C C C G G G G G G C C C
XmaI  (1)
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  (3)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRV  (8)
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.
BpmI  (13)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
PmlI  (13)
1 site
C A C G T G G T G C A C
HindIII  (17)
2 sites
A A G C T T T T C G A A
BspQI  (307)
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  (307)
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  (423)
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  (423)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (427)
1 site
R C A T G Y Y G T A C R
BssSαI  (596)
1 site
C A C G A G G T G C T C
BseYI  (727)
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.
PspFI  (731)
1 site
C C C A G C G G G T C G
ApaLI  (737)
1 site
G T G C A C C A C G T G
BaeGI  (741)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (741)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
BsiHKAI  (741)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
AlwNI  (839)
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.
AcuI  (971)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PflMI  (1436)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
KanR
1313 .. 2131  =  819 bp
273 codons
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1313 .. 2131  =  819 bp
273 codons
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
484 .. 1072  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
484 .. 1072  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2367 .. 2822  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2367 .. 2822  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
2237 .. 2341  =  105 bp
AmpR promoter
2237 .. 2341  =  105 bp
MCS
3021 .. 22  =  42 bp
multiple cloning site
MCS
3021 .. 22  =  42 bp
multiple cloning site
lac promoter
130 .. 160  =  31 bp
3 segments
   Segment 3:  -10  
   130 .. 136  =  7 bp
promoter for the E. coli lac operon
lac promoter
130 .. 160  =  31 bp
3 segments
   Segment 2:  
   137 .. 154  =  18 bp
promoter for the E. coli lac operon
lac promoter
130 .. 160  =  31 bp
3 segments
   Segment 1:  -35  
   155 .. 160  =  6 bp
promoter for the E. coli lac operon
lac promoter
130 .. 160  =  31 bp
3 segments
promoter for the E. coli lac operon
T3 promoter
43 .. 61  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
43 .. 61  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T7 promoter
2990 .. 3008  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2990 .. 3008  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
82 .. 98  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
82 .. 98  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
106 .. 122  =  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
106 .. 122  =  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).
M13 fwd
2964 .. 2980  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2964 .. 2980  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1313 .. 1924  =  612 bp
ORF:  203 amino acids  =  23.4 kDa
ORF:  2605 .. 86  =  522 bp
ORF:  173 amino acids  =  18.8 kDa
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