pSTBlue-1

Multi-purpose bacterial cloning and expression vector with a versatile MCS, T7 and SP6 promoters, and ampicillin and kanamycin resistance genes.

Sequence Author: MilliporeSigma (Novagen)

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Acc65I (51) M13 rev lac operator BspQI - SapI (3627) PciI (3510) PspFI (3210) BseYI (3206) AlwNI (3101) PflMI (2660) Bpu10I (2414) BsmBI (2413) AsiSI (2397) EcoNI (2309) SmaI (2271) TspMI - XmaI (2269) BspDI - ClaI (2088) NruI (2054) KpnI (55) SphI (63) PstI (68) MluI (70) SnaBI (78) BamHI (84) EcoRI (91) EcoRV (101) EcoRI (106) SalI (112) AccI (113) HincII (114) HindIII (118) PaeR7I - XhoI (124) AvrII - StyI (130) NheI (135) BmtI (139) XbaI (141) AleI - PmlI (152) BstXI (154) EcoO109I - PspOMI (159) ApaI (163) Eco53kI (167) SacI (169) EagI - NotI (172) SP6 promoter AanI - PsiI (469) DraIII (597) BtgZI (598) NgoMIV (698) NaeI (700) BsaHI (1207) TatI (1264) ScaI (1266) NmeAIII (1600) BpmI (1678) BsaI (1681) AhdI (1747) pSTBlue-1 3851 bp
Acc65I  (51)
1 site
G G T A C C C C A T G G
BspQI  (3627)
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  (3627)
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.
PciI  (3510)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (3210)
1 site
C C C A G C G G G T C G
BseYI  (3206)
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.
AlwNI  (3101)
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.
PflMI  (2660)
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.
Bpu10I  (2414)
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  (2413)
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.
AsiSI  (2397)
1 site
G C G A T C G C C G C T A G C G
EcoNI  (2309)
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.
SmaI  (2271)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (2269)
1 site
C C C G G G G G G C C C
XmaI  (2269)
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.
BspDI  (2088)
1 site
A T C G A T T A G C T A
ClaI  (2088)
1 site
A T C G A T T A G C T A
NruI  (2054)
1 site
T C G C G A A G C G C T
KpnI  (55)
1 site
G G T A C C C C A T G G
SphI  (63)
1 site
G C A T G C C G T A C G
PstI  (68)
1 site
C T G C A G G A C G T C
MluI  (70)
1 site
A C G C G T T G C G C A
SnaBI  (78)
1 site
T A C G T A A T G C A T
BamHI  (84)
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.
EcoRI  (91)
2 sites
G A A T T C C T T A A G
EcoRV  (101)
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  (106)
2 sites
G A A T T C C T T A A G
SalI  (112)
1 site
G T C G A C C A G C T G
AccI  (113)
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  (114)
1 site
G T Y R A C C A R Y T G
HindIII  (118)
1 site
A A G C T T T T C G A A
PaeR7I  (124)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (124)
1 site
C T C G A G G A G C T C
AvrII  (130)
1 site
C C T A G G G G A T C C
StyI  (130)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NheI  (135)
1 site
G C T A G C C G A T C G
BmtI  (139)
1 site
G C T A G C C G A T C G
XbaI  (141)
1 site
T C T A G A A G A T C T
AleI  (152)
1 site
C A C N N N N G T G G T G N N N N C A C
PmlI  (152)
1 site
C A C G T G G T G C A C
BstXI  (154)
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.
EcoO109I  (159)
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.
PspOMI  (159)
1 site
G G G C C C C C C G G G
ApaI  (163)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
Eco53kI  (167)
1 site
G A G C T C C T C G A G
SacI  (169)
1 site
G A G C T C C T C G A G
EagI  (172)
1 site
C G G C C G G C C G G C
NotI  (172)
1 site
G C G G C C G C C G C C G G C G
AanI  (469)
1 site
T T A T A A A A T A T T
PsiI  (469)
1 site
T T A T A A A A T A T T
DraIII  (597)
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  (598)
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  (698)
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  (700)
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.
BsaHI  (1207)
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.
TatI  (1264)
1 site
W G T A C W W C A T G W
ScaI  (1266)
1 site
A G T A C T T C A T G A
NmeAIII  (1600)
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  (1678)
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  (1681)
1 site
G G T C T C N C C A G A G <