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Plasmid Files

pFA6a-GST-kanMX6

Plasmid with a kanMX marker for adding a C-terminal GST tag.

 
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pFA6a-GST-kanMX6.dna
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PvuII (15) NdeI (4732) PfoI (4594) AatII (4483) ZraI (4481) NmeAIII (3709) BpmI (3631) BmrI (3601) BanI (3509) AlwNI (3084) PspFI (2976) BseYI (2972) HpaI (2489) SacII (2437) SfiI (2430) SpeI (2417) EcoRV (2407) BsiWI (25) SalI (37) F2 (42 .. 61) BamHI (43) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) BtgZI (181) MscI (270) BsgI (349) BstBI (460) SwaI (490) BclI * (497) AscI - BssHII (739) PsiI (800) BstZ17I (817) BglII (947) BstEII (977) BstXI (994) BmgBI (1030) BseRI (1141) MluI (1194) NcoI - StyI (1334) NruI (1418) AsiSI (1761) PflMI (2024) PmeI (2382) Eco53kI (2389) SacI (2391) EcoRI (2393) R1 (2379 .. 2398) pFA6a-GST-kanMX6 4815 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
NdeI  (4732)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
PfoI  (4594)
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.
AatII  (4483)
1 site
G A C G T C C T G C A G
ZraI  (4481)
1 site
G A C G T C C T G C A G
NmeAIII  (3709)
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).
BpmI  (3631)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BmrI  (3601)
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  (3509)
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  (3084)
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  (2976)
1 site
C C C A G C G G G T C G
BseYI  (2972)
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.
HpaI  (2489)
1 site
G T T A A C C A A T T G
SacII  (2437)
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.
SfiI  (2430)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SpeI  (2417)
1 site
A C T A G T T G A T C A
EcoRV  (2407)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
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
BtgZI  (181)
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.
MscI  (270)
1 site
T G G C C A A C C G G T
BsgI  (349)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BstBI  (460)
1 site
T T C G A A A A G C T T
SwaI  (490)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BclI  (497)
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.
AscI  (739)
1 site
G G C G C G C C C C G C G C G G
BssHII  (739)
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.
PsiI  (800)
1 site
T T A T A A A A T A T T
BstZ17I  (817)
1 site
G T A T A C C A T A T G
BglII  (947)
1 site
A G A T C T T C T A G A
BstEII  (977)
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.
BstXI  (994)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BmgBI  (1030)
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  (1141)
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  (1194)
1 site
A C G C G T T G C G C A
NcoI  (1334)
1 site
C C A T G G G G T A C C
StyI  (1334)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NruI  (1418)
1 site
T C G C G A A G C G C T
AsiSI  (1761)
1 site
G C G A T C G C C G C T A G C G
PflMI  (2024)
1 site
C C A