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Plasmid Files

pFA6a-His3MX6-PGAL1

Plasmid for swapping in the GAL1 promoter using the HIS3MX6 selectable marker.

 
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pFA6a-His3MX6-PGAL1.dna
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BsiWI (25) HindIII (19) NdeI (4246) PfoI (4108) EcoO109I (4051) AatII (3997) ZraI (3995) SspI (3879) XmnI (3674) ScaI (3555) PvuI (3445) BmrI (3115) BanI (3023) AlwNI (2598) PspFI (2490) BseYI (2486) SalI (37) BamHI (43) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) R2 (65 .. 84) BstAPI (209) AgeI (454) BglII (617) BstEII (647) BmgBI (700) Bpu10I (709) MluI (864) NcoI - StyI (1004) PsiI (1117) SphI (1132) BclI * (1263) BsaBI (1265) NgoMIV (1333) NaeI (1335) XbaI (1384) AfeI (1572) BssHII (1599) PmeI (1896) Eco53kI (1903) SacI (1905) EcoRI (1907) F4 (1893 .. 1912) EcoRV (1921) SfiI (1944) SacII (1951) HpaI (2003) BspQI - SapI (2066) pFA6a-His3MX6-PGAL1 4329 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.
HindIII  (19)
1 site
A A G C T T T T C G A A
NdeI  (4246)
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  (4108)
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.
EcoO109I  (4051)
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.
AatII  (3997)
1 site
G A C G T C C T G C A G
ZraI  (3995)
1 site
G A C G T C C T G C A G
SspI  (3879)
1 site
A A T A T T T T A T A A
XmnI  (3674)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3555)
1 site
A G T A C T T C A T G A
PvuI  (3445)
1 site
C G A T C G G C T A G C
BmrI  (3115)
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  (3023)
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  (2598)
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  (2490)
1 site
C C C A G C G G G T C G
BseYI  (2486)
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.
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
BstAPI  (209)
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  (454)
1 site
A C C G G T T G G C C A
BglII  (617)
1 site
A G A T C T T C T A G A
BstEII  (647)
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.
BmgBI  (700)
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.
Bpu10I  (709)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
MluI  (864)
1 site
A C G C G T T G C G C A
NcoI  (1004)
1 site
C C A T G G G G T A C C
StyI  (1004)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PsiI  (1117)
1 site
T T A T A A A A T A T T
SphI  (1132)
1 site
G C A T G C C G T A C G
BclI  (1263)
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.
BsaBI  (1265)
1 site
G A T N N N N A T C C T A N N N N T A G
NgoMIV  (1333)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (1335)
1 site
G C C G G C C G G C C G