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Plasmid Files

pFA6a-TRP1-PGAL1-3HA

Plasmid with a TRP1 marker for swapping in the GAL1 promoter and adding a triple-HA tag.

 
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pFA6a-TRP1-PGAL1-3HA.dna
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PvuII (15) EcoO109I (3997) SspI (3825) ScaI (3501) PvuI (3391) FspI (3243) BpmI (3091) BanI (2969) AlwNI (2544) AflIII - PciI (2128) BsiWI (25) PstI (35) SalI (37) PsiI (194) AscI - BssHII (251) BbvCI - Bpu10I (257) R3 (261 .. 280) BamHI (297) PacI (371) BtgZI (541) AgeI (767) BseRI (805) BglII (930) BsrGI (935) BspEI * (948) PmlI (1043) XbaI (1167) MfeI (1223) Bsu36I (1325) BstXI (1381) BsgI (1496) AarI (1516) PmeI (1842) Eco53kI (1849) SacI (1851) EcoRI (1853) F4 (1839 .. 1858) BspDI - ClaI (1860) SpeI (1877) SfiI (1890) BtgI (1894) SacII (1897) HpaI (1949) BspQI - SapI (2012) pFA6a-TRP1-PGAL1-3HA 4275 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
EcoO109I  (3997)
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  (3825)
1 site
A A T A T T T T A T A A
ScaI  (3501)
1 site
A G T A C T T C A T G A
PvuI  (3391)
1 site
C G A T C G G C T A G C
FspI  (3243)
1 site
T G C G C A A C G C G T
BpmI  (3091)
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.
BanI  (2969)
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  (2544)
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.
AflIII  (2128)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2128)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
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.
PstI  (35)
1 site
C T G C A G G A C G T C
SalI  (37)
1 site
G T C G A C C A G C 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
Bpu10I  (257)
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.
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
BtgZI  (541)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
AgeI  (767)
1 site
A C C G G T T G G C C A
BseRI  (805)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BglII  (930)
1 site
A G A T C T T C T A G A
BsrGI  (935)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BspEI  (948)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PmlI  (1043)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
XbaI  (1167)
1 site
T C T A G A A G A T C T
MfeI  (1223)
1 site
C A A T T G G T T A A C
Bsu36I  (1325)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BstXI  (1381)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BsgI  (1496)
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).
AarI  (1516)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PmeI  (1842)
1 site
G T T T A A A C C A A A T T T G
Eco53kI  (1849)
1 site
G A G C T C C T C G A G
SacI  (1851)
1 site
G A G C T C C T C G A G
EcoRI  (1853)
1 site
G A A T T C C T T A A G
BspDI  (1860)
1 site
A T C G A T T A G C T A
ClaI  (1860)
1 site
A T C G A T T A G C T A
SpeI  (1877)
1 site
A C T A G T T G A T C A
SfiI  (1890)
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.
BtgI  (1894)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (1897)
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.
HpaI  (1949)
1 site
G T T A A C C A A T T G
BspQI  (2012)
1 site
G C T C T T C N C G A