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pNIC-CTHF

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

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pNIC-CTHF.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Structural Genomics Consortium
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BglII (7241) SgrAI (7200) SphI (7052) BstAPI (6843) MluI (6519) BclI * (6505) BstEII (6337) NmeAIII (6319) ApaI (6316) PspOMI (6312) BssHII (6108) PshAI (5678) BglI (5462) FspI - FspAI (5441) PpuMI (5413) PflFI - Tth111I (4676) BspQI - SapI (4535) AlwNI (4009) NruI (3563) XbaI (47) AflII (69) BfuAI (81) StuI * (770) MscI (792) AanI - AanI (1169) BsrGI (1209) SnaBI (1278) Acc65I (1586) KpnI (1590) SacII (1608) ZraI (1989) AatII (1991) PstI (2008) AarI - BfuAI (2011) BamHI (2075) EcoRI (2081) Eco53kI (2089) SacI (2091) SalI (2094) HindIII (2100) EagI - NotI (2107) PaeR7I - PspXI - XhoI (2115) BlpI (2194) DraIII (2522) AanI - AanI (2647) AsiSI - PvuI (3222) TspMI - XmaI (3344) SmaI (3346) pNIC-CTHF 7260 bp
BglII  (7241)
1 site
A G A T C T T C T A G A
SgrAI  (7200)
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  (7052)
1 site
G C A T G C C G T A C G
BstAPI  (6843)
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  (6519)
1 site
A C G C G T T G C G C A
BclI  (6505)
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  (6337)
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  (6319)
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  (6316)
1 site
G G G C C C C