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Plasmid Files

pUAST

P element-based vector for Gal4-regulated expression of genes in Drosophila.

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pUAST Sequence and MappUAST.dna
Map and Sequence File   
Sequence Author:  Brand Lab
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 SbfI (8529) SphI (8523) Bpu10I (8397) white linker sequence NdeI (7733) AatII (7484) ZraI (7482) FspI (6784) BsaI (6623) AhdI (6562) EcoNI (5169) PpuMI (4992) NsiI (4789) BsrGI (4620) EcoRI (1) BglII (13) EagI - NotI (20) MCS SacII (26) PaeR7I - PspXI - XhoI (29) Acc65I (35) KpnI (39) XbaI (41) small t intron MfeI (597) PsiI (630) SV40 poly(A) signal StuI (760) AfeI (962) AflII (1345) KasI (1961) NarI (1962) SfoI (1963) PluTI (1965) NcoI (2310) BtgZI (2523) Eco53kI (2635) SacI (2637) SnaBI (2825) TspMI - XmaI (2917) SmaI (2919) BstXI (2951) NruI (3259) BspDI - ClaI (3504) EcoRV (3749) pUAST 8897 bp
SbfI  (8529)
1 site
C C T G C A G G G G A C G T C C
SphI  (8523)
1 site
G C A T G C C G T A C G
Bpu10I  (8397)
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.
NdeI  (7733)
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.
AatII  (7484)
1 site
G A C G T C C T G C A G
ZraI  (7482)
1 site
G A C G T C C T G C A G
FspI  (6784)
1 site
T G C G C A A C G C G T
BsaI  (6623)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (6562)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
EcoNI  (5169)
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.
PpuMI  (4992)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
NsiI  (4789)
1 site
A T G C A T T A C G T A
BsrGI  (4620)
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.
EcoRI  (1)
1 site
G A A T T C C T T A A G
BglII  (13)
1 site
A G A T C T T C T A G A
EagI  (20)
1 site
C G G C C G G C C G G C
NotI  (20)
1 site
G C G G C C G C C G C C G G C G
SacII  (26)
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.
PaeR7I  (29)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (29)
1 site
V C T C G A G B B G A G C T C V
XhoI  (29)
1 site
C T C G A G G A G C T C
Acc65I  (35)
1 site
G G T A C C C C A T G G
KpnI  (39)
1 site
G G T A C C C C A T G G
XbaI  (41)
1 site
T C T A G A A G A T C T
MfeI  (597)
1 site
C A A T T G G T T A A C
PsiI  (630)
1 site
T T A T A A A A T A T T
StuI  (760)
1 site
A G G C C T T C C G G A
AfeI  (962)
1 site
A G C G C T T C G C G A
AflII  (1345)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
KasI  (1961)
1 site
G G C G C C C C G C G G
NarI  (1962)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (1963)
1 site
G G C G C C C C G C G G
PluTI  (1965)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
NcoI  (2310)
1 site
C C A T G G G G T A C C
BtgZI  (2523)
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.
Eco53kI  (2635)
1 site
G A G C T C C T C G A G
SacI  (2637)
1 site
G A G C T C C T C G A G
SnaBI  (2825)
1 site
T A C G T A A T G C A T
TspMI  (2917)
1 site
C C C G G G G G G C C C
XmaI  (2917)
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.
SmaI  (2919)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BstXI  (2951)
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.
NruI  (3259)
1 site
T C G C G A A G C G C T
BspDI  (3504)
1 site
A T C G A T T A G C T A
ClaI  (3504)
1 site
A T C G A T T A G C T A
EcoRV  (3749)
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.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 1:  
   1250 .. 1321  =  72 bp
   24 amino acids  =  2.7 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 2:  
   1716 .. 1989  =  274 bp
   91 amino acids  =  9.8 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 3:  
   2064 .. 2718  =  655 bp
   219 amino acids  =  23.8 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 4:  
   2780 .. 3095  =  316 bp
   105 amino acids  =  12.1 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 5:  
   3316 .. 3447  =  132 bp
   44 amino acids  =  4.8 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
   Segment 6:  
   3518 .. 4132  =  615 bp
   204 amino acids  =  22.5 kDa
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
mini-white
1250 .. 4132  =  2883 bp
687 amino acids  =  75.7 kDa
6 segments
Product: Drosophila white gene eye color pigment
This is a modified version of the white gene lacking
part of the first intron.
AmpR
6489 .. 7349  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   6489 .. 7280  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6489 .. 7349  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   7281 .. 7349  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6489 .. 7349  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
5730 .. 6318  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5730 .. 6318  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
P element 5' end
4910 .. 5495  =  586 bp
   Segment 1:  
   4910 .. 5464  =  555 bp
P element 5' end
4910 .. 5495  =  586 bp
   Segment 2:  inverted repeat  
   5465 .. 5495  =  31 bp
P element 5' end
4910 .. 5495  =  586 bp
2 segments
white linker sequence
7755 .. 8255  =  501 bp
white linker sequence
white linker sequence
7755 .. 8255  =  501 bp
white linker sequence
hsp70 promoter
8646 .. 8884  =  239 bp
Drosophila melanogaster hsp70Bb promoter
hsp70 promoter
8646 .. 8884  =  239 bp
Drosophila melanogaster hsp70Bb promoter
P element 3' end
8256 .. 8488  =  233 bp
   Segment 1:  inverted repeat  
   8256 .. 8286  =  31 bp
P element 3' end
P element 3' end
8256 .. 8488  =  233 bp
   Segment 2:  
   8287 .. 8488  =  202 bp
P element 3' end
P element 3' end
8256 .. 8488  =  233 bp
2 segments
P element 3' end
white linker sequence
5496 .. 5724  =  229 bp
white linker sequence
white linker sequence
5496 .. 5724  =  229 bp
white linker sequence
SV40 poly(A) signal
611 .. 745  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
611 .. 745  =  135 bp
SV40 polyadenylation signal
AmpR promoter
7350 .. 7454  =  105 bp
AmpR promoter
7350 .. 7454  =  105 bp
5X UAS
8533 .. 8627  =  95 bp
five tandem copies of the "ScaI site" 17-mer
CGGAGTACTGTCCTCCG, an upstream activating
sequence (UAS) that efficiently binds yeast Gal4
(Webster et al., 1988; Pfeiffer et al., 2010)
5X UAS
8533 .. 8627  =  95 bp
five tandem copies of the "ScaI site" 17-mer
CGGAGTACTGTCCTCCG, an upstream activating
sequence (UAS) that efficiently binds yeast Gal4
(Webster et al., 1988; Pfeiffer et al., 2010)
small t intron
124 .. 189  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
124 .. 189  =  66 bp
simian virus 40 (SV40) small t antigen intron
MCS
1 .. 46  =  46 bp
multiple cloning site
MCS
1 .. 46  =  46 bp
multiple cloning site
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