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pMCSG40 (linearized)

Linearized bacterial vector for ligation-independent cloning (LIC), with E. coli DsbA plus OsmY fused to a TVMV-6xHis-TEV leader

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pMCSG40 (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
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ScaI (5882) PvuI (5772) BsaI (5463) AhdI (5402) PciI (4509) BspQI - SapI (4393) BstZ17I (4280) BsaAI (4261) PpuMI (3515) FspAI (3490) StyI (6634) AvaI - BsoBI - PaeR7I - PspXI - XhoI (6735) BmeT110I (6736) EagI - NotI (6743) HindIII (6750) SalI (6756) Eco53kI (6765) SacI (6767) EcoRI (6769) BamHI (6775) End (6781) Start (0) BsrGI (102) DraIII (182) BstBI (684) NdeI (710) XbaI (748) BspDI * - ClaI * (817) SgrAI (859) AgeI (1347) AscI (1358) BglII (1515) BfuAI - BspMI (1639) MfeI (1741) EcoNI (1943) PflMI (1990) BstAPI (2091) MluI (2408) BclI * (2422) PspOMI (2615) ApaI (2619) pMCSG40 6781 bp
ScaI  (5882)
1 site
A G T A C T T C A T G A
PvuI  (5772)
1 site
C G A T C G G C T A G C
BsaI  (5463)
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  (5402)
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.
PciI  (4509)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4393)
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  (4393)
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.
BstZ17I  (4280)
1 site
G T A T A C C A T A T G
BsaAI  (4261)
1 site
Y A C G T R R T G C A Y
PpuMI  (3515)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
FspAI  (3490)
1 site
R T G C G C A Y Y A C G C G T R
StyI  (6634)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
AvaI  (6735)
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  (6735)
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.
PaeR7I  (6735)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6735)
1 site
V C T C G A G B B G A G C T C V
XhoI  (6735)
1 site
C T C G A G G A G C T C
BmeT110I  (6736)
1 site
C Y C G R G G R G C Y C
EagI  (6743)
1 site
C G G C C G G C C G G C
NotI  (6743)
1 site
G C G G C C G C C G C C G G C G
HindIII  (6750)
1 site
A A G C T T T T C G A A
SalI  (6756)
1 site
G T C G A C C A G C T G
Eco53kI  (6765)
1 site
G A G C T C C T C G A G
SacI  (6767)
1 site
G A G C T C C T C G A G
EcoRI  (6769)
1 site
G A A T T C C T T A A G
BamHI  (6775)
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.
End  (6781)
0 sites
Start  (0)
0 sites
BsrGI  (102)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
DraIII  (182)
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.
BstBI  (684)
1 site
T T C G A A A A G C T T
NdeI  (710)
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.
XbaI  (748)
1 site
T C T A G A A G A T C T
BspDI  (817)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (817)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (859)
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.
AgeI  (1347)
1 site
A C C G G T T G G C C A
AscI  (1358)
1 site
G G C G C G C C C C G C G C G G
BglII  (1515)
1 site
A G A T C T T C T A G A
BfuAI  (1639)
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.