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Plasmid Files

pMMB66HE

Broad-host-range bacterial cloning vector. See also pMMB66EH.

 
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pMMB66HE.dna
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HindIII (5) PacI (3) MauBI (8543) MluI (8086) ApaI - BanII (7883) PspOMI (7879) HpaI (7584) BsmBI (7471) BspQI - SapI (7278) FspAI (6807) BsaAI (6752) EcoNI (6657) PfoI * (6517) SgrAI (4569) PstI (17) SalI (19) BamHI (25) TspMI - XmaI (30) SmaI (32) EcoRI (35) BssS α I (682) DraI (773) PvuI (982) AhdI (1351) SexAI * (1475) SfiI (1709) EcoO109I (2261) BstZ17I (3084) PasI (4090) pMMB66HE 8822 bp
HindIII  (5)
1 site
A A G C T T T T C G A A
PacI  (3)
1 site
T T A A T T A A A A T T A A T T
MauBI  (8543)
1 site
C G C G C G C G G C G C G C G C
MluI  (8086)
1 site
A C G C G T T G C G C A
ApaI  (7883)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (7883)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (7879)
1 site
G G G C C C C C C G G G
HpaI  (7584)
1 site
G T T A A C C A A T T G
BsmBI  (7471)
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.
BspQI  (7278)
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  (7278)
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.
FspAI  (6807)
1 site
R T G C G C A Y Y A C G C G T R
BsaAI  (6752)
1 site
Y A C G T R R T G C A Y
EcoNI  (6657)
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.
PfoI  (6517)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SgrAI  (4569)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PstI  (17)
1 site
C T G C A G G A C G T C
SalI  (19)
1 site
G T C G A C C A G C T G
BamHI  (25)
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.
TspMI  (30)
1 site
C C C G G G G G G C C C
XmaI  (30)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (32)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (35)
1 site
G A A T T C C T T A A G
BssSαI  (682)
1 site
C A C G A G G T G C T C
DraI  (773)
1 site
T T T A A A A A A T T T
PvuI  (982)
1 site
C G A T C G G C T A G C
AhdI  (1351)
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.
SexAI  (1475)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (1709)
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.
EcoO109I  (2261)
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.
BstZ17I  (3084)
1 site
G T A T A C C A T A T G