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pMCSG62

Bacterial vector encoding tRNA genes for rare Arg and Ile codons, for expressing a protein with a 6xHis-AviTag™-TEV leader plus the BirA biotin ligase.

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pMCSG62.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
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BspEI (2) ScaI (5174) FspI (4916) BglI (4814) BsaI (4755) AhdI (4694) DrdI (3909) PciI (3801) BspQI - SapI (3685) BstZ17I (3572) PshAI (3546) BsmBI (3316) HpaI (3207) EcoRV (3151) ApaI (2912) BlpI (80) AvaI - BsoBI - PaeR7I - PspXI - XhoI (158) BmeT110I (159) EagI - NotI (166) HindIII (173) SalI (179) Eco53kI (188) SacI (190) EcoRI (192) BamHI (198) BsmI (651) DraIII (1079) SspI (1247) Acc65I (1272) KpnI (1276) NdeI (1348) XbaI (1386) BspDI * - ClaI * (1455) AgeI (1497) AscI (1714) SbfI (1725) BfuAI - BspMI (1743) MreI (1862) AfeI (1948) NcoI (2132) PsiI (2147) EcoNI (2236) PfoI (2268) PflMI (2283) BstAPI (2384) MluI (2701) BclI * (2715) BstEII (2882) PspOMI (2908) pMCSG62 5869 bp
BspEI  (2)
1 site
T C C G G A A G G C C T
ScaI  (5174)
1 site
A G T A C T T C A T G A
FspI  (4916)
1 site
T G C G C A A C G C G T
BglI  (4814)
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  (4755)
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  (4694)
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  (3909)
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  (3801)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3685)
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  (3685)
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  (3572)
1 site
G T A T A C C A T A T G
PshAI  (3546)
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  (3316)
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  (3207)
1 site
G T T A A C C A A T T G
EcoRV  (3151)
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  (2912)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BlpI  (80)
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  (158)
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  (158)
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  (158)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site
V C T C G A G B B G A G C T C V
XhoI  (158)
1 site
C T C G A G G A G C T C
BmeT110I  (159)
1 site
C Y C G R G G R G C Y C
EagI  (166)
1 site
C G G C C G G C C G G C
NotI  (166)
1 site
G C G G C C G C C G C C G G C G
HindIII  (173)
1 site
A A G C T T T T C G A A
SalI  (179)
1 site
G T C G A C C A G C T G
Eco53kI  (188)
1 site
G A G C T C C T C G A G
SacI  (190)
1 site
G A G C T C C T C G A G
EcoRI  (192)
1 site
G A A T T C C T T A A G
BamHI  (198)
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  (651)
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  (1079)
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.
SspI  (1247)
1 site
A A T A T T T T A T A A
Acc65I  (1272)
1 site
G G T A C C C C A T G G
KpnI  (1276)
1 site
G G T A C C C C A T G G
NdeI  (1348)
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  (1386)
1 site
T C T A G A A G A T C T
BspDI  (1455)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1455)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (1497)
1 site
A C C G G