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Plasmid Files

pFA6a-His3MX6-PGAL1-3HA

Plasmid with a HIS3MX6 marker for swapping in the GAL1 promoter and adding a triple-HA tag.

 
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pFA6a-His3MX6-PGAL1-3HA.dna
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HindIII (19) EcoO109I (4364) SspI (4192) XmnI (3987) ScaI (3868) PvuI (3758) BmrI (3428) BanI (3336) AlwNI (2911) PspFI (2803) BseYI (2799) BspQI - SapI (2379) HpaI (2316) SacII (2264) BsiWI (25) SalI (37) BstZ17I (177) AscI (251) BbvCI (257) R3 (261 .. 280) BamHI (297) PacI (371) BstAPI (522) AgeI (767) BglII (930) BstEII (960) BmgBI (1013) MluI (1177) NcoI - StyI (1317) SphI (1445) BclI * (1576) BsaBI (1578) NgoMIV (1646) NaeI (1648) XbaI (1697) AfeI (1885) PmeI (2209) Eco53kI (2216) SacI (2218) EcoRI (2220) F4 (2206 .. 2225) EcoRV (2234) SfiI (2257) pFA6a-His3MX6-PGAL1-3HA 4642 bp
HindIII  (19)
1 site
A A G C T T T T C G A A
EcoO109I  (4364)
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  (4192)
1 site
A A T A T T T T A T A A
XmnI  (3987)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3868)
1 site
A G T A C T T C A T G A
PvuI  (3758)
1 site
C G A T C G G C T A G C
BmrI  (3428)
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  (3336)
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  (2911)
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  (2803)
1 site
C C C A G C G G G T C G
BseYI  (2799)
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.
BspQI  (2379)
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  (2379)
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  (2316)
1 site
G T T A A C C A A T T G
SacII  (2264)
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.
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
BbvCI  (257)
1 site
C C T C A G C G