pBS(-)

Phagemid cloning vector, also known as BlueScribe M13 Minus or BlueM13m or pBSM13(–).
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EcoO109I (3193) AatII (3139) ZraI (3137) XmnI (2816) ScaI (2697) NmeAIII (2365) BpmI (2287) BsaI (2278) AhdI (2217) AlwNI (1740) PspFI (1632) PfoI (46) NdeI (184) BstAPI (185) NgoMIV (364) NaeI (366) BtgZI (464) BsaAI (469) DraIII (472) PsiI (597) T7 promoter EcoRI (881) Eco53kI (889) SacI (891) Acc65I (893) AvaI - BsoBI - KpnI - TspMI - XmaI (897) SmaI (899) BamHI (902) XbaI (908) SalI (914) AccI (915) HincII (916) PstI - SbfI (924) BfuAI - BspMI (927) SphI (930) HindIII (932) T3 promoter lac operator BspQI - SapI (1208) AflIII - PciI (1324) BseYI (1628) pBS(-) 3204 bp
EcoO109I  (3193)
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.
AatII  (3139)
1 site
G A C G T C C T G C A G
ZraI  (3137)
1 site
G A C G T C C T G C A G
XmnI  (2816)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2697)
1 site
A G T A C T T C A T G A
NmeAIII  (2365)
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).
BpmI  (2287)
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.
BsaI  (2278)
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  (2217)
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  (1740)
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  (1632)
1 site
C C C A G C G G G T C G
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  (184)
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.
BstAPI  (185)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NgoMIV  (364)
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  (366)
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  (464)
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  (469)
1 site
Y A C G T R R T G C A Y
DraIII  (472)
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  (597)
1 site
T T A T A A A A T A T T
EcoRI  (881)
1 site
G A A T T C C T T A A G
Eco53kI  (889)
1 site
G A G C T C C T C G A G
SacI  (891)
1 site
G A G C T C C T C G A G
Acc65I  (893)
1 site
G G T A C C C C A T G G
AvaI  (897)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (897)
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.
KpnI  (897)
1 site
G G T A C C C C A T G G
TspMI  (897)
1 site
C C C G G G G G G C C C
XmaI  (897)
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  (899)
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  (902)
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.
XbaI  (908)
1 site
T C T A G A A G A T C T
SalI  (914)
1 site
G T C G A C C A G C T G
AccI  (915)
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  (916)
1 site
G T Y R A C C A R Y T G
PstI  (924)
1 site
C T G C A G G A C G T C
SbfI  (924)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (927)
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  (927)
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.
SphI  (930)
1 site
G C A T G C C G T A C G
HindIII  (932)
1 site
A A G C T T T T C G A A
BspQI  (1208)
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  (1208)
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  (1324)
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  (1324)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1628)
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
2144 .. 3004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2144 .. 2935  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2144 .. 3004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2936 .. 3004  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2144 .. 3004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1385 .. 1973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1385 .. 1973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
239 .. 694  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
239 .. 694  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
3005 .. 3109  =  105 bp
AmpR promoter
3005 .. 3109  =  105 bp
MCS
881 .. 937  =  57 bp
pUC18/19 multiple cloning site
MCS
881 .. 937  =  57 bp
pUC18/19 multiple cloning site
lac promoter
1031 .. 1061  =  31 bp
3 segments
   Segment 3:  -10  
   1031 .. 1037  =  7 bp
promoter for the E. coli lac operon
lac promoter
1031 .. 1061  =  31 bp
3 segments
   Segment 2:  
   1038 .. 1055  =  18 bp
promoter for the E. coli lac operon
lac promoter
1031 .. 1061  =  31 bp
3 segments
   Segment 1:  -35  
   1056 .. 1061  =  6 bp
promoter for the E. coli lac operon
lac promoter
1031 .. 1061  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
860 .. 878  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
860 .. 878  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
944 .. 962  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
944 .. 962  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
837 .. 853  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
837 .. 853  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
983 .. 999  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
983 .. 999  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
1007 .. 1023  =  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
1007 .. 1023  =  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).
lacZα
679 .. 987  =  309 bp
102 amino acids  =  11.7 kDa
Product: LacZα fragment of β-galactosidase
lacZα
679 .. 987  =  309 bp
102 amino acids  =  11.7 kDa
Product: LacZα fragment of β-galactosidase
ORF:  2274 .. 2540  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  679 .. 987  =  309 bp
ORF:  102 amino acids  =  11.7 kDa
ORF:  2144 .. 3004  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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