pMAL-c6T

Bacterial vector for inducible cytoplasmic expression of maltose-binding protein (MBP) fusions with a TEV cleavage site.

Sequence Author: New England Biolabs

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EcoO109I * - PpuMI * (5225) Bpu10I (5216) PflFI - Tth111I (4891) BspQI - SapI (4810) PciI (4693) Bsu36I (4004) BglI (3770) FspI (3665) ScaI (3407) DraI (3310) BspHI (3048) RsrII (3004) HindIII (2771) PstI - SbfI (2759) EcoRI (2748) BamHI (2742) AccI (2737) SalI - SgrDI (2736) EcoRV (2732) EagI - NotI (2723) AlwNI (2714) AvaI - BsoBI (2688) SacI (2655) PflMI (12) BstAPI (113) MluI (430) BstEII (611) PspOMI (637) ApaI (641) HpaI (936) KasI (1069) NarI * (1070) SfoI (1071) PluTI (1073) lac operator MfeI (1501) ATG BspEI (1664) BsiWI (1835) PsiI (1897) BglII (1904) BmgBI (2087) BsmI (2165) BsaBI (2220) BtgI (2232) BlpI (2331) AfeI (2455) Eco53kI (2653) pMAL-c6T 5247 bp
EcoO109I  (5225)
1 site
R G G N C C Y Y C C N G G R
* Blocked by Dcm methylation.
Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (5225)
1 site
R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
Bpu10I  (5216)
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.
PflFI  (4891)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4891)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BspQI  (4810)
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  (4810)
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  (4693)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
Bsu36I  (4004)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BglI  (3770)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
FspI  (3665)
1 site
T G C G C A A C G C G T
ScaI  (3407)
1 site
A G T A C T T C A T G A
DraI  (3310)
1 site
T T T A A A A A A T T T
BspHI  (3048)
1 site
T C A T G A A G T A C T
RsrII  (3004)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
HindIII  (2771)
1 site
A A G C T T T T C G A A
PstI  (2759)
1 site
C T G C A G G A C G T C
SbfI  (2759)
1 site
C C T G C A G G G G A C G T C C
EcoRI  (2748)
1 site
G A A T T C C T T A A G
BamHI  (2742)
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.
AccI  (2737)
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.
SalI  (2736)
1 site
G T C G A C C A G C T G
SgrDI  (2736)
1 site
C G T C G A C G G C A G C T G C
EcoRV  (2732)
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.
EagI  (2723)
1 site
C G G C C G G C C G G C
NotI  (2723)
1 site
G C G G C C G C C G C C G G C G
AlwNI  (2714)
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.
AvaI  (2688)
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  (2688)
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.
SacI  (2655)
1 site
G A G C T C C T C G A G
PflMI  (12)
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.
BstAPI  (113)
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.
MluI  (430)
1 site
A C G C G T T G C G C A
BstEII  (611)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PspOMI  (637)
1 site
G G G C C C C C C G G G
ApaI  (641)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
HpaI  (936)
1 site
G T T A A C C A A T T G
KasI  (1069)
1 site
G G C G C C C C G C G G
NarI  (1070)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1071)
1 site
G G C G C C C C G C G G
PluTI  (1073)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
MfeI  (1501)
1 site
C A A T T G G T T A A C
BspEI  (1664)
1 site
T C C G G A