pNYCOMPS-LIC-FH10T+ (N term)

Bacterial vector for high-throughput expression and purification of membrane proteins with N-terminal FLAG® and 10xHis tags.

Sequence Author: New York Structural Biology Center

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SnaBI (6840) EagI - NotI (6798) EcoRI (6478) PasI (6247) ScaI (6065) BbvCI (5754) BsrGI (5634) SrfI (5611) BmgBI (5577) BsaI (5469) BfuAI - BspMI (5363) PstI (5360) SalI (5350) SnaBI (5313) TEV site FLAG AanI (5240) ATG RBS XbaI (5192) T7 promoter SgrAI (5085) SphI (4937) BstAPI (4728) MluI (4404) BclI * (4390) BstEII (4222) NmeAIII (4204) ApaI (4201) PspOMI (4197) EcoRV (3958) HpaI (3902) PshAI (3563) BglI (3347) PaeR7I - PspXI - XhoI (0) BlpI (79) DraIII (407) AanI (532) AsiSI - PvuI (1107) BspDI - ClaI (1412) NruI (1448) AcuI (1761) BssSI (2130) PciI (2303) BspQI - SapI (2420) PflFI - Tth111I (2561) PpuMI (3298) FspI - FspAI (3326) pNYCOMPS-LIC-FH10T+ (N term) 6858 bp
SnaBI  (6840)
2 sites
T A C G T A A T G C A T
EagI  (6798)
1 site
C G G C C G G C C G G C
NotI  (6798)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (6478)
1 site
G A A T T C C T T A A G
PasI  (6247)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
ScaI  (6065)
1 site
A G T A C T T C A T G A
BbvCI  (5754)
1 site
C C T C A G C G G A G T C G
BsrGI  (5634)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SrfI  (5611)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (5577)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BsaI  (5469)
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.
BfuAI  (5363)
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  (5363)
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.
PstI  (5360)
1 site
C T G C A G G A C G T C
SalI  (5350)
1 site
G T C G A C C A G C T G
SnaBI  (5313)
2 sites
T A C G T A A T G C A T
AanI  (5240)
2 sites
T T A T A A A A T A T T
XbaI  (5192)
1 site
T C T A G A A G A T C T
SgrAI  (5085)
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  (4937)
1 site
G C A T G C C G T A C G
BstAPI  (4728)
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  (4404)
1 site
A C G C G T T G C G C