Resources
Plasmid Files

pFA6a-kanMX6-PGAL1

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

 
To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

 PvuII (15) NdeI (4402) PfoI (4264) EcoO109I (4207) AatII (4153) ZraI (4151) XmnI (3830) BpmI (3301) BmrI (3271) BanI (3179) AlwNI (2754) PspFI (2646) BseYI (2642) PciI (2338) BspQI - SapI (2222) BsiWI (25) SalI (37) AccI (38) BamHI (43) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) R2 (65 .. 84) BstAPI (209) BtgZI (228) AgeI (454) BglII (617) BstEII (647) BstXI (664) BmgBI (700) MluI (864) NcoI - StyI (1004) NruI (1088) EcoNI (1343) AsiSI (1431) PflMI (1694) PmeI (2052) Eco53kI (2059) SacI (2061) EcoRI (2063) F4 (2049 .. 2068) EcoRV (2077) SpeI (2087) SfiI (2100) SacII (2107) HpaI (2159) pFA6a-kanMX6-PGAL1 4485 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
NdeI  (4402)
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  (4264)
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  (4207)
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  (4153)
1 site
G A C G T C C T G C A G
ZraI  (4151)
1 site
G A C G T C C T G C A G
XmnI  (3830)
1 site
G A A N N N N T T C C T T N N N N A A G
BpmI  (3301)
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  (3271)
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  (3179)
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  (2754)
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  (2646)
1 site
C C C A G C G G G T C G
BseYI  (2642)
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  (2338)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2222)
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  (2222)
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.
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
AccI  (38)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
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.
BtgZI  (228)
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  (454)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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.
BstXI  (664)
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.
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.
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.
NruI  (1088)
1 site
T C G C G A A G C G C T
EcoNI  (1343)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
AsiSI  (1431)
1 site
G C G A T C G C C G C T A G C G
PflMI  (1694)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
PmeI  (2052)
1 site
G T T T A A A C C A A A T T T G
Eco53kI  (2059)
1 site
G A G C T C C T C G A G
SacI  (2061)
1 site
G A G C T C C T C G A G
EcoRI  (2063)
1 site
G A A T T C C T T A A G
EcoRV  (2077)
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.
SpeI  (2087)
1 site
A C T A G T T G A T C A
SfiI  (2100)
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.
SacII  (2107)
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  (2159)
1 site
G T T A A C C A A T T G
R2
20-mer  /  40% GC
1 binding site
65 .. 84  =  20 annealed bases
Tm  =  53°C
Reverse primer for promoter swapping with the
GAL1 promoter. This primer encodes the reverse
complement of a start codon. A gene-specific
sequence should be added at the 5' end of the
primer.
F4
20-mer  /  40% GC
1 binding site
2049 .. 2068  =  20 annealed bases
Tm  =  52°C
Forward primer for promoter swapping. This primer
includes an EcoRI recognition sequence. A
gene-specific sequence should be added at the 5'
end of the primer.
kanMX
662 .. 2018  =  1357 bp
yeast selectable marker conferring kanamycin
resistance
kanMX
662 .. 2018  =  1357 bp
yeast selectable marker conferring kanamycin
resistance
AmpR
3158 .. 4018  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3158 .. 3949  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3158 .. 4018  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3950 .. 4018  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3158 .. 4018  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2399 .. 2987  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
2399 .. 2987  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
GAL1 promoter
87 .. 528  =  442 bp
inducible promoter, regulated by Gal4
GAL1 promoter
87 .. 528  =  442 bp
inducible promoter, regulated by Gal4
AmpR promoter
4019 .. 4123  =  105 bp
AmpR promoter
4019 .. 4123  =  105 bp
T7 promoter
2123 .. 2141  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2123 .. 2141  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
4469 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
4469 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
KanR
1006 .. 1815  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
1006 .. 1815  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
TEF promoter
662 .. 1005  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
662 .. 1005  =  344 bp
Ashbya gossypii TEF promoter
TEF terminator
1821 .. 2018  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
1821 .. 2018  =  198 bp
Ashbya gossypii TEF terminator
UAS
411 .. 528  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
UAS
411 .. 528  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter