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Plasmid Files

pFA6a-His3MX6-PGAL1-GST

Plasmid with a HIS3MX6 marker for swapping in the GAL1 promoter and adding a GST tag.

 
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pFA6a-His3MX6-PGAL1-GST.dna
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HindIII (19) NdeI (5126) PfoI (4988) AatII (4877) ZraI (4875) SspI (4759) PvuI (4325) BmrI (3995) BanI (3903) AlwNI (3478) PspFI (3370) BseYI (3366) HpaI (2883) SacII (2831) SfiI (2824) EcoRV (2801) F4 (2773 .. 2792) EcoRI (2787) SacI (2785) Eco53kI (2783) BsiWI (25) SalI (37) BstZ17I (177) AscI (251) R4 (261 .. 280) SwaI (504) BstBI (532) BsgI (647) MscI (724) EcoNI (920) PacI (938) BstAPI (1089) AgeI (1334) BglII (1497) BstEII (1527) BmgBI (1580) Bpu10I (1589) MluI (1744) NcoI - StyI (1884) SphI (2012) BsaBI (2145) NgoMIV (2213) NaeI (2215) XbaI (2264) AfeI (2452) PmeI (2776) pFA6a-His3MX6-PGAL1-GST 5209 bp
HindIII  (19)
1 site
A A G C T T T T C G A A
NdeI  (5126)
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  (4988)
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  (4877)
1 site
G A C G T C C T G C A G
ZraI  (4875)
1 site
G A C G T C C T G C A G
SspI  (4759)
1 site
A A T A T T T T A T A A
PvuI  (4325)
1 site
C G A T C G G C T A G C
BmrI  (3995)
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  (3903)
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  (3478)
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  (3370)
1 site
C C C A G C G G G T C G
BseYI  (3366)
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  (2883)
1 site
G T T A A C C A A T T G
SacII  (2831)
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  (2824)
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.
EcoRV  (2801)
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  (2787)
1 site
G A A T T C C T T A A G
SacI  (2785)
1 site
G A G C T C C T C G A G
Eco53kI  (2783)
1 site
G A G C T C C T C G A 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
AscI  (251)
1 site
G G C G C G C C C C G C G C G G
SwaI  (504)
1 site
A T T T A A A T T A A