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Plasmid Files

pFA6a-GFP(S65T)-His3MX6

Plasmid with a HIS3MX6 marker for adding a C-terminal GFP tag or truncating a gene.

 
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pFA6a-GFP(S65T)-His3MX6.dna
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HindIII (19) PfoI (4477) EcoO109I (4420) AatII (4366) ZraI (4364) SspI (4248) XmnI (4043) ScaI (3924) PvuI (3814) NmeAIII (3592) BmrI (3484) BanI (3392) BspQI - SapI (2435) SacII (2320) BsiWI (25) SalI (37) F2 (42 .. 61) BamHI (43) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) MscI (230) SnaBI (253) BsrGI (335) PmlI (384) MfeI (619) BstBI (681) F3 (777 .. 793) AscI (778) BglII (986) BstEII (1016) BmgBI (1069) Bpu10I (1078) BseRI (1180) MluI (1233) SphI (1501) BclI * (1632) BsaBI (1634) NgoMIV (1702) NaeI (1704) XbaI (1753) AfeI (1941) PmeI (2265) Eco53kI (2272) BanII - SacI (2274) EcoRI (2276) R1 (2262 .. 2281) EcoRV (2290) SfiI (2313) pFA6a-GFP(S65T)-His3MX6 4698 bp
HindIII  (19)
1 site
A A G C T T T T C G A A
PfoI  (4477)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
EcoO109I  (4420)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (4366)
1 site
G A C G T C C T G C A G
ZraI  (4364)
1 site
G A C G T C C T G C A G
SspI  (4248)
1 site
A A T A T T T T A T A A
XmnI  (4043)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3924)
1 site
A G T A C T T C A T G A
PvuI  (3814)
1 site
C G A T C G G C T A G C
NmeAIII  (3592)
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).
BmrI  (3484)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
BanI  (3392)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
BspQI  (2435)
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  (2435)
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.
SacII  (2320)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BsiWI  (25)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
SalI  (37)
1 site
G T C G A C C A G C T G
BamHI  (43)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
AvaI  (48)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (48)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (48)
1 site
C C C G G G G G G C C C
XmaI  (48)
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.
BmeT110I  (49)
1 site
C Y C G R G G R G C Y C
SmaI  (50)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PacI  (58)
1 site
T T A A T T A A A A T T A A T T
MscI  (230)
1 site
T G G C C A A C C G G T
SnaBI  (253)
1 site
T A C G T A A T G C A T
BsrGI  (335)
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.
PmlI  (384)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
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