pMCSG62 (linearized)

Linearized bacterial vector for ligation-independent cloning (LIC), with a 6xHis-AviTag™-TEV leader plus the BirA biotin ligase plus tRNA genes for rare Arg and Ile codons.

Sequence Author: Midwest Center for Structural Genomics

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BspEI (4624) ScaI (3927) FspI (3669) BglI (3567) BsaI (3508) AhdI (3447) DrdI (2662) PciI (2554) BspQI - SapI (2438) BstZ17I (2325) PshAI (2299) BsmBI - Esp3I (2069) HpaI (1960) EcoRV (1904) ApaI (1665) PspOMI (1661) BstEII (1635) BlpI (4702) AvaI - BsoBI - PaeR7I - PspXI - XhoI (4780) EagI - NotI (4788) HindIII (4795) SalI (4801) Eco53kI (4810) SacI (4812) EcoRI (4814) BamHI (4820) BsmI (5273) DraIII (5701) End (5854) Start (0) TEV site Acc65I (25) KpnI (29) 6xHis ATG NdeI (101) RBS XbaI (139) T7 promoter BspDI * - ClaI * (208) AgeI (250) AscI (467) SbfI (478) BfuAI - BspMI (496) MreI (615) AfeI (701) NcoI (885) PsiI (900) EcoNI (989) PfoI (1021) PflMI (1036) BstAPI (1137) MluI (1454) BclI * (1468) pMCSG62 5854 bp
BspEI  (4624)
1 site
T C C G G A A G G C C T
ScaI  (3927)
1 site
A G T A C T T C A T G A
FspI  (3669)
1 site
T G C G C A A C G C G T
BglI  (3567)
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.
BsaI  (3508)
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  (3447)
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.
DrdI  (2662)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PciI  (2554)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2438)
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  (2438)
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  (2325)
1 site
G T A T A C C A T A T G
PshAI  (2299)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BsmBI  (2069)
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.
Esp3I  (2069)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
HpaI  (1960)
1 site
G T T A A C C A A T T G
EcoRV  (1904)
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.
ApaI  (1665)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (1661)
1 site
G G G C C C C C C G G G
BstEII  (1635)
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.
BlpI  (4702)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
AvaI  (4780)
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  (4780)
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  (4780)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4780)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4780)
1 site
C T C G A G G A G C T C
EagI  (4788)
1 site
C G G C C G G C C G G C
NotI  (4788)
1 site
G C G G C C G C C G C C G G C G
HindIII  (4795)
1 site
A A G C T T T T C G A A
SalI  (4801)
1 site
G T C G A C C A G C T G
Eco53kI  (4810)
1 site
G A G C T C C T C G A G
SacI  (4812)
1 site
G A G C T C C T C G A G
EcoRI  (4814)
1 site
G A A T T C C T T A A G
BamHI  (4820)
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.
BsmI  (5273)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
DraIII  (5701)
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.
End  (5854)
0 sites
Start  (0)
0 sites
Acc65I  (25)
1 site
G G T A C C C C A T G G
KpnI  (29)
1 site
G G T A C C C C A T G G
NdeI  (101)
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  (139)
1 site
T C T A G A A G A T C T
BspDI  (208)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (208)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (250)
1 site
A C C G G T T G G C C A
AscI  (467)
1 site
G G C G C G C C C C G C G C G G
SbfI  (478)
1 site
C C T G C A G G G G A C G T C C