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Plasmid Files

pFA6a-3HA-His3MX6

Plasmid with a HIS3MX6 marker for adding a C-terminal triple-HA tag.

 
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pFA6a-3HA-His3MX6.dna
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BsiWI (25) HindIII (19) EcoO109I (3814) SspI (3642) XmnI (3437) ScaI (3318) PvuI (3208) NmeAIII (2986) BmrI (2878) BanI (2786) AlwNI (2361) PspFI (2253) BseYI (2249) SalI (37) F2 (42 .. 61) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) BbvCI (167) AscI (172) BstZ17I (250) BglII (380) BstEII (410) BmgBI (463) BseRI (574) MluI (627) NcoI - StyI (767) BtgZI (810) SphI (895) BclI * (1026) BsaBI (1028) NgoMIV (1096) NaeI (1098) XbaI (1147) AfeI (1335) PmeI (1659) Eco53kI (1666) BanII - SacI (1668) EcoRI (1670) R1 (1656 .. 1675) EcoRV (1684) SfiI (1707) SacII (1714) HpaI (1766) BspQI - SapI (1829) pFA6a-3HA-His3MX6 4092 bp
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.
HindIII  (19)
1 site
A A G C T T T T C G A A
EcoO109I  (3814)
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.
SspI  (3642)
1 site
A A T A T T T T A T A A
XmnI  (3437)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3318)
1 site
A G T A C T T C A T G A
PvuI  (3208)
1 site
C G A T C G G C T A G C
NmeAIII  (2986)
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  (2878)
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  (2786)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
AlwNI  (2361)
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.
PspFI  (2253)
1 site
C C C A G C G G G T C G
BseYI  (2249)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
SalI  (37)
1 site
G T C G A C C A G C T G
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
BbvCI  (167)
1 site
C C T C A G C G G A G T C G
AscI  (172)
1 site
G G C G C G C C C C G C G C G G
BstZ17I  (250)
1 site
G T A T A C C A T A T G
BglII  (380)
1 site
A G A T C T T C T A G A
BstEII  (410)
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.
BmgBI  (463)
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.
BseRI  (574)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
MluI  (627)
1 site
A C G C G T T G C G C A
NcoI  (767)
1 site
C C A T G G G G T A C C
StyI  (767)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BtgZI  (810)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SphI  (895)
1 site
G C A T G C C G T A C G
BclI  (1026)
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.
BsaBI  (1028)
1 site
G A T N N N N A T C C T A N N N N T A G
NgoMIV  (1096)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (1098)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
XbaI  (1147)
1 site
T C T A G A A G A T C T
AfeI  (1335)
1 site
A G C G C T T C G C G A
PmeI  (1659)
1 site
G T T T A A A C C A A A T T T G
Eco53kI  (1666)
1 site
G A G C T C C T C G A G
BanII  (1668)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (1668)
1 site
G A G C T C C T C G A G
EcoRI  (1670)
1 site
G A A T T C C T T A A G
EcoRV  (1684)
1 site