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pMCSG75

Bacterial vector encoding tRNA genes for rare Arg and Ile codons, plus a C-terminal TEV-AviTag™-6xHis cassette, plus the BirA biotin ligase.

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pMCSG75.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
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BspEI (2) ScaI (5181) FspI (4923) BglI (4821) BsaI (4762) AhdI (4701) DrdI (3916) PciI (3808) BspQI - SapI (3692) BstZ17I (3579) PshAI (3553) BsmBI (3323) HpaI (3214) EcoRV (3158) ApaI (2919) BlpI (80) PaeR7I - PspXI - XhoI (158) EagI - NotI (166) HindIII (173) SalI (179) Eco53kI (188) SacI (190) EcoRI (192) BsmI (651) DraIII (1079) TspMI - XmaI (1354) SmaI (1356) XbaI (1393) BspDI * - ClaI * (1462) AgeI (1504) AscI (1721) SbfI (1732) BfuAI - BspMI (1750) MreI (1869) AfeI (1955) NcoI (2139) PsiI (2154) EcoNI (2243) PfoI (2275) PflMI (2290) BstAPI (2391) MluI (2708) BclI * (2722) BstEII (2889) PspOMI (2915) pMCSG75 5876 bp
BspEI  (2)
1 site
T C C G G A A G G C C T
ScaI  (5181)
1 site
A G T A C T T C A T G A
FspI  (4923)
1 site
T G C G C A A C G C G T
BglI  (4821)
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  (4762)
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  (4701)
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  (3916)
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  (3808)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3692)
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  (3692)
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  (3579)
1 site
G T A T A C C A T A T G
PshAI  (3553)
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  (3323)
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  (3214)
1 site
G T T A A C C A A T T G
EcoRV  (3158)
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  (2919)
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.
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
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
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.
TspMI  (1354)
1 site
C C C G G G G G G C C C
XmaI  (1354)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1356)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
XbaI  (1393)
1 site
T C T A G A A G A T C T
BspDI  (1462)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1462)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (1504)
1 site
A C C G G T T G G C C A
AscI  (1721)
1 site
G G C G C G C C C C G C G C G G
SbfI  (1732)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (1750)
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.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1750)
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 BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
MreI  (1869)
1 site
C G C C G G C G G C G G C C G C
AfeI  (1955)
1 site
A G C G C T T C G C G A
NcoI  (2139)
1 site
C C A T G G G G T A C C
PsiI  (2154)
1 site
T T A T A A A A T A T T
EcoNI  (2243)
1 site