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Plasmid Files

pNIC-CH

Bacterial vector encoding a C-terminal 6xHis-tag plus a SacB negative selection marker, for purification of recombinant proteins.

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pNIC-CH.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Structural Genomics Consortium
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BstAPI (7053) MluI (6729) BclI * (6715) BstEII (6547) NmeAIII (6529) ApaI (6526) PspOMI (6522) BssHII (6318) PshAI (5888) BglI (5672) FspI - FspAI (5651) PpuMI (5623) PflFI - Tth111I (4886) BspQI - SapI (4745) AlwNI (4219) NruI (3773) SmaI (3556) SphI (44) SgrAI (192) BglII (233) XbaI (299) AflII (321) BfuAI (333) StuI * (1022) MscI (1044) AanI - AanI (1421) BsrGI (1461) SnaBI (1530) Acc65I (1838) KpnI (1842) SacII (1860) ZraI (2241) AatII (2243) PstI (2260) AarI - BfuAI (2263) BamHI (2285) EcoRI (2291) Eco53kI (2299) SacI (2301) SalI (2304) HindIII (2310) EagI - NotI (2317) PaeR7I - PspXI - XhoI (2325) BlpI (2404) DraIII (2732) AanI - AanI (2857) AsiSI - PvuI (3432) TspMI - XmaI (3554) pNIC-CH 7218 bp
BstAPI  (7053)
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  (6729)
1 site
A C G C G T T G C G C A
BclI  (6715)
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  (6547)
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  (6529)
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  (6526)
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  (6522)
1 site
G G G C C C C C C G G G
BssHII  (6318)
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.
PshAI  (5888)
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.
BglI  (5672)
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  (5651)
1 site
T G C G C A A C G C G T
FspAI  (5651)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (5623)
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  (4886)
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  (4886)
1 site
G A C N N N G T C