pFA6a-kanMX6-PGAL1-GST

Plasmid with a kanMX marker for swapping in the GAL1 promoter and adding a GST tag.
|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Yeast Plasmids | More Plasmid Sets
No matches
PvuII (15) NdeI (5282) PfoI (5144) AatII (5033) ZraI (5031) BpmI (4181) BmrI (4151) BanI (4059) AlwNI (3634) PspFI (3526) BseYI (3522) HpaI (3039) SacII (2987) SfiI (2980) SpeI (2967) EcoRV (2957) F4 (2929 .. 2948) EcoRI (2943) SacI (2941) Eco53kI (2939) PmeI (2932) BsiWI (25) SalI (37) BstZ17I (177) PsiI (194) AscI - BssHII (251) R4 (261 .. 280) BclI * (493) SwaI (504) BstBI (532) BsgI (647) MscI (724) PacI (938) BstAPI (1089) AgeI (1334) BglII (1497) BstEII (1527) BstXI (1544) BmgBI (1580) MluI (1744) NcoI - StyI (1884) NruI (1968) AsiSI (2311) PflMI (2574) pFA6a-kanMX6-PGAL1-GST 5365 bp
PvuII  (15)
1 site
C A G C T G G T C G A C
NdeI  (5282)
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  (5144)
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.
AatII  (5033)
1 site
G A C G T C C T G C A G
ZraI  (5031)
1 site
G A C G T C C T G C A G
BpmI  (4181)
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  (4151)
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  (4059)
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  (3634)
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  (3526)
1 site
C C C A G C G G G T C G
BseYI  (3522)
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.
HpaI  (3039)
1 site
G T T A A C C A A T T G
SacII  (2987)
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  (2980)
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  (2967)
1 site
A C T A G T T G A T C A
EcoRV  (2957)
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.
EcoRI  (2943)
1 site
G A A T T C C T T A A G
SacI  (2941)
1 site
G A G C T C C T C G A G
Eco53kI  (2939)
1 site
G A G C T C C T C G A G
PmeI  (2932)
1 site
G T T T A A A C C A A A T T T G
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
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.
BclI  (493)
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.
SwaI  (504)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BstBI  (532)
1 site
T T C G A A A A G C T T
BsgI  (647)
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).
MscI  (724)
1 site
T G G C C A A C C G G T
PacI  (938)
1 site
T T A A T T A A A A T T A A T T
BstAPI  (1089)
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  (1334)
1 site
A C C G G T T G G C C A
BglII  (1497)
1 site
A G A T C T T C T A G A
BstEII  (1527)
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  (1544)
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  (1580)
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  (1744)
1 site
A C G C G T T G C G C A
NcoI  (1884)
1 site
C C A T G G G G T A C C
StyI  (1884)
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  (1968)
1 site
T C G C G A A G C G C T
AsiSI  (2311)
1 site
G C G A T C G C C G C T A G C G
PflMI  (2574)
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.
F4
20-mer  /  40% GC
1 binding site
2929 .. 2948  =  20 annealed bases
Tm  =  53°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.
R4
20-mer  /  55% GC
1 binding site
261 .. 280  =  20 annealed bases
Tm  =  62°C
Reverse primer for promoter swapping and adding an N-terminal GST tag. A gene-specific sequence should be added at the 5' end of the primer.
kanMX
1542 .. 2898  =  1357 bp
yeast selectable marker conferring kanamycin resistance
kanMX
1542 .. 2898  =  1357 bp
yeast selectable marker conferring kanamycin resistance
AmpR
4038 .. 4898  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4038 .. 4829  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4038 .. 4898  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4830 .. 4898  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4038 .. 4898  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
GST
279 .. 932  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
279 .. 932  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
start codon
945 .. 947  =  3 bp
1 amino acid  =  149.2 Da
start codon
945 .. 947  =  3 bp
1 amino acid  =  149.2 Da
ori
3279 .. 3867  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
3279 .. 3867  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
GAL1 promoter
967 .. 1408  =  442 bp
inducible promoter, regulated by Gal4
GAL1 promoter
967 .. 1408  =  442 bp
inducible promoter, regulated by Gal4
ADH1 terminator
50 .. 237  =  188 bp
ADH1 terminator
50 .. 237  =  188 bp
AmpR promoter
4899 .. 5003  =  105 bp
AmpR promoter
4899 .. 5003  =  105 bp
T7 promoter
3003 .. 3021  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3003 .. 3021  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
5349 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
5349 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
KanR
1886 .. 2695  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1886 .. 2695  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
TEF promoter
1542 .. 1885  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
1542 .. 1885  =  344 bp
Ashbya gossypii TEF promoter
TEF terminator
2701 .. 2898  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
2701 .. 2898  =  198 bp
Ashbya gossypii TEF terminator
UAS
1291 .. 1408  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
UAS
1291 .. 1408  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
ORF:  1483 .. 1752  =  270 bp
ORF:  89 amino acids  =  10.2 kDa
ORF:  4168 .. 4434  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1886 .. 2695  =  810 bp
ORF:  269 amino acids  =  30.7 kDa
ORF:  258 .. 947  =  690 bp
ORF:  229 amino acids  =  26.7 kDa
ORF:  4038 .. 4898  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
Click here to try SnapGene

Download pFA6a-kanMX6-PGAL1-GST.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page 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.

Discover the most user-friendly molecular biology experience.