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Plasmid Files

pFA6a-kanMX6-PGAL1-GST

Plasmid with a kanMX marker for swapping in the GAL1 promoter and adding a GST tag.

 
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pFA6a-kanMX6-PGAL1-GST.dna
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PvuII (15) NdeI (5282) PfoI (5144) AatII (5033) ZraI (5031) BpmI (4181) BmrI (4151) BanI (4059) AlwNI (3634) PspFI (3526) BseYI (3522) HpaI (3039) SacII (2987) SfiI (2980) SpeI (2967) EcoRV (2957) F4 (2929 .. 2948) EcoRI (2943) SacI (2941) Eco53kI (2939) PmeI (2932) BsiWI (25) SalI (37) BstZ17I (177) PsiI (194) AscI - BssHII (251) R4 (261 .. 280) BclI * (493) SwaI (504) BstBI (532) BsgI (647) MscI (724) PacI (938) BstAPI (1089) AgeI (1334) BglII (1497) BstEII (1527) BstXI (1544) BmgBI (1580) MluI (1744) NcoI - StyI (1884) NruI (1968) AsiSI (2311) PflMI (2574) pFA6a-kanMX6-PGAL1-GST 5365 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
NdeI  (5282)
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  (5144)
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  (5033)
1 site
G A C G T C C T G C A G
ZraI  (5031)
1 site
G A C G T C C T G C A G
BpmI  (4181)
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  (4151)
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  (4059)
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  (3634)
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  (3526)
1 site
C C C A G C G G G T C G
BseYI  (3522)
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  (3039)
1 site
G T T A A C C A A T T G
SacII  (2987)
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  (2980)
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  (2967)
1 site
A C T A G T T G A T C A
EcoRV  (2957)
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.
EcoRI  (2943)
1 site
G A A T T C C T T A A G
SacI  (2941)
1 site
G A G C T C C T C G A G
Eco53kI  (2939)
1 site
G A G C T C C T C G A G
PmeI  (2932)
1 site
G T T T A A A C C A A A T T T G
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
BstZ17I  (177)
1 site
G T A T A C C A T A T G
PsiI  (194)
1 site
T T A T A A A A T A T T
AscI  (251)
1 site
G G C G C G C C C C G C G C G G
BssHII  (251)
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.
BclI  (493)
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.
SwaI  (504)
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.
BstBI  (532)
1 site
T T C G A A A A G C T T
BsgI  (647)
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).
MscI  (724)
1 site
T G G C C A A C C G G T
PacI  (938)
1 site
T T A A T T A A A A T T A A T T
BstAPI  (1089)
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.
AgeI  (1334)
1 site
A C C G G T T G G C C A
BglII  (1497)
1 site
A G A T C T T C T A G A
B