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pNIC28-Bsa4

Bacterial vector encoding an N-terminal 6xHis-TEV cassette and kanamycin resistance plus a SacB negative selection marker, for purification of recombinant proteins.

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pNIC28-Bsa4.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Structural Genomics Consortium
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SgrAI (7224) SphI (7076) BstAPI (6867) MluI (6543) BclI * (6529) BstEII (6361) NmeAIII (6343) ApaI (6340) PspOMI (6336) BssHII (6132) BglI (5486) FspI - FspAI (5465) PpuMI (5437) PflFI - Tth111I (4700) BspQI - SapI (4559) AlwNI (4033) NruI (3587) XbaI (47) NdeI (87) BsaI (152) NcoI (153) ZraI (166) AatII (168) BfuAI - BspMI (186) StuI * (857) MscI (879) AanI - AanI (1256) BsrGI (1296) SnaBI (1365) SacII (1695) BsaI (2083) BamHI (2099) EcoRI (2105) Eco53kI (2113) SacI (2115) SalI (2118) HindIII (2124) EagI - NotI (2131) PaeR7I - PspXI - XhoI (2139) BlpI (2218) DraIII (2546) AanI - AanI (2671) AsiSI - PvuI (3246) TspMI - XmaI (3368) SmaI (3370) pNIC28-Bsa4 7284 bp
SgrAI  (7224)
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.
SphI  (7076)
1 site
G C A T G C C G T A C G
BstAPI  (6867)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
MluI  (6543)
1 site
A C G C G T T G C G C A
BclI  (6529)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (6361)
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.
NmeAIII  (6343)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ApaI  (6340)
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  (6336)
1 site
G G G C C C C C C G G G
BssHII  (6132)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BglI  (5486)
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.
FspI  (5465)
1 site
T G C G C A A C G C G T
FspAI  (5465)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (5437)
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.
PflFI  (4700)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4700)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BspQI  (4559)
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  (4559)
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.
AlwNI  (4033)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
NruI  (3587)
1 site
T C G C G A A G C G C T
XbaI  (47)
1 site
T C T A G A A G A T C T
NdeI  (87)
1 site
C A T A T G G T A T