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Plasmid Files

pFA6a-kanMX6-PGAL1-3HA

Plasmid with a kanMX marker for swapping in the GAL1 promoter and adding a triple-HA tag.

 
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pFA6a-kanMX6-PGAL1-3HA.dna
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BsiWI (25) PvuII (15) EcoO109I (4520) XmnI (4143) BpmI (3614) BmrI (3584) BanI (3492) AlwNI (3067) PspFI (2959) BseYI (2955) PciI (2651) BspQI - SapI (2535) HpaI (2472) SacII (2420) SfiI (2413) SpeI (2400) SalI (37) BstZ17I (177) PsiI (194) AscI - BssHII (251) BbvCI (257) R3 (261 .. 280) BamHI (297) PacI (371) BstAPI (522) BtgZI (541) AgeI (767) BglII (930) BstEII (960) BstXI (977) BmgBI (1013) MluI (1177) NcoI - StyI (1317) NruI (1401) EcoNI (1656) AsiSI (1744) PflMI (2007) PmeI (2365) Eco53kI (2372) SacI (2374) EcoRI (2376) F4 (2362 .. 2381) EcoRV (2390) pFA6a-kanMX6-PGAL1-3HA 4798 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.
PvuII  (15)
1 site
C A G C T G G T C G A C
EcoO109I  (4520)
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.
XmnI  (4143)
1 site
G A A N N N N T T C C T T N N N N A A G
BpmI  (3614)
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  (3584)
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  (3492)
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  (3067)
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  (2959)
1 site
C C C A G C G G G T C G
BseYI  (2955)
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.
PciI  (2651)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2535)
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  (2535)
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.
HpaI  (2472)
1 site
G T T A A C C A A T T G
SacII  (2420)
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  (2413)
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  (2400)
1 site
A C T A G T T G A T C A
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.
BbvCI  (257)
1 site
C C T C A G C G G A G T C G
BamHI  (297)
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.
PacI  (371)
1 site
T T A A T T A A A A T T A A T T
BstAPI  (522)
1 site
G C A N N N N N T G C C G T N N N N N A C G