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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.

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pMCSG62 (linearized).dna
Map and Sequence File:    Download    Open   
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 (2069) HpaI (1960) EcoRV (1904) ApaI (1665) PspOMI (1661) BstEII (1635) BlpI (4702) AvaI - BsoBI - PaeR7I - PspXI - XhoI (4780) BmeT110I (4781) EagI - NotI (4788) HindIII (4795) SalI (4801) Eco53kI (4810) SacI (4812) EcoRI (4814) BamHI (4820) BsmI (5273) DraIII (5701) End (5854) Start (0) Acc65I (25) KpnI (29) NdeI (101) XbaI (139) 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.
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