pMCSG31

Bacterial expression vector with an MBP-TVMV-6xHis-TEV leader and a Tet-inducible TVMV protease cassette.

Sequence Author: Midwest Center for Structural Genomics

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BsgI (4952) AleI (4928) NruI (4670) SnaBI (4627) EcoNI (4598) Bpu10I (4551) AhdI (4307) BpmI (4238) NmeAIII (4160) MfeI (4129) FspI (4084) ScaI (3826) XmnI (3707) BstBI (2834) PflMI (7) BseRI (246) AgeI (411) BsmBI - Esp3I (552) BbsI (596) AflIII - MluI (910) SphI (1031) SgrAI (1179) BspDI * - ClaI * (1223) T7 promoter RBS PfoI * (1588) BsiWI (1622) BglII (1691) BstXI (1717) BssHII (1768) BmgBI (1874) BsrGI (2449) 6xHis Acc65I (2526) KpnI (2530) XcmI (2553) SspI (2555) Eco53kI (2586) SacI (2588) EagI - NotI (2604) TspMI - XmaI (2768) SmaI (2770) AbsI (2779) SbfI (2795) Strep-Tag II pMCSG31 5822 bp
BsgI  (4952)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AleI  (4928)
1 site
C A C N N N N G T G G T G N N N N C A C
NruI  (4670)
1 site
T C G C G A A G C G C T
SnaBI  (4627)
1 site
T A C G T A A T G C A T
EcoNI  (4598)
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.
Bpu10I  (4551)
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.
AhdI  (4307)
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.
BpmI  (4238)
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.
NmeAIII  (4160)
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).
MfeI  (4129)
1 site
C A A T T G G T T A A C
FspI  (4084)
1 site
T G C G C A A C G C G T
ScaI  (3826)
1 site
A G T A C T T C A T G A
XmnI  (3707)
1 site
G A A N N N N T T C C T T N N N N A A G
BstBI  (2834)
1 site
T T C G A A A A G C T T
PflMI  (7)
1 site
C C A N N N N N T G G G G T N N N