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Plasmid Files

pME18S-FL3

Mammalian vector for directional cloning and transient expression of a cDNA.

 
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 HindIII (1) SspI (3277) BsaHI (3010) ScaI (2953) TatI (2951) FspI (2695) BpmI (2543) AhdI (2473) DrdI (1688) AarI (4) SexAI * (99) BtgI - NcoI (239) SfiI (285) AvrII (332) EcoNI (401) Bsu36I (402) Bpu10I (496) EcoO109I - PpuMI (692) PfoI (694) EcoRI (794) XhoI (800) DraIII (811) EcoRV (816) AccI (960) AleI (1094) DraIII (1194) XhoI (1198) PstI (1208) EagI - NotI (1214) SpeI (1224) XbaI (1229) MfeI (1283) PsiI (1316) TspMI - XmaI (1432) SmaI (1434) Acc65I (1436) KpnI (1440) Eco53kI (1444) SacI (1446) AflIII - PciI (1580) pME18S-FL3 3392 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
SspI  (3277)
1 site
A A T A T T T T A T A A
BsaHI  (3010)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
ScaI  (2953)
1 site
A G T A C T T C A T G A
TatI  (2951)
1 site
W G T A C W W C A T G W
FspI  (2695)
1 site
T G C G C A A C G C G T
BpmI  (2543)
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.
AhdI  (2473)
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.
DrdI  (1688)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AarI  (4)
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.
SexAI  (99)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BtgI  (239)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (239)
1 site
C C A T G G G G T A C C
SfiI  (285)
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.
AvrII  (332)
1 site
C C T A G G G G A T C C
EcoNI  (401)
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.
Bsu36I  (402)
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.
Bpu10I  (496)
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.
EcoO109I  (692)
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.
PpuMI  (692)
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.
PfoI  (694)
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.
EcoRI  (794)
1 site
G A A T T C C T T A A G
XhoI  (800)
2 sites
C T C G A G G A G C T C
DraIII  (811)
2 sites
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
EcoRV  (816)
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.
AccI  (960)
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.
AleI  (1094)
1 site
C A C N N N N G T G G T G N N N N C A C
DraIII  (1194)
2 sites
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
XhoI  (1198)
2 sites
C T C G A G G A G C T C
PstI  (1208)
1 site
C T G C A G G A C G T C
EagI  (1214)
1 site
C G G C C G G C C G G C
NotI  (1214)
1 site
G C G G C C G C C G C C G G C G
SpeI  (1224)
1 site
A C T A G T T G A T C A
XbaI  (1229)
1 site
T C T A G A A G A T C T
MfeI  (1283)
1 site
C A A T T G G T T A A C
PsiI  (1316)
1 site
T T A T A A A A T A T T
TspMI  (1432)
1 site
C C C G G G G G G C C C
XmaI  (1432)
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  (1434)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (1436)
1 site
G G T A C C C C A T G G
KpnI  (1440)
1 site
G G T A C C C C A T G G
Eco53kI  (1444)
1 site
G A G C T C C T C G A G
SacI  (1446)
1 site
G A G C T C C T C G A G
AflIII  (1580)
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  (1580)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2400 .. 3260  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2400 .. 3191  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2400 .. 3260  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3192 .. 3260  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2400 .. 3260  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
SRα promoter
18 .. 632  =  615 bp
   Segment 1:  SV40 promoter  
   18 .. 347  =  330 bp
hybrid promoter consisting of the SV40 early
promoter plus part of the long terminal repeat of
human T-lymphotrophic virus 1
SRα promoter
18 .. 632  =  615 bp
   Segment 2:  
   348 .. 363  =  16 bp
hybrid promoter consisting of the SV40 early
promoter plus part of the long terminal repeat of
human T-lymphotrophic virus 1
SRα promoter
18 .. 632  =  615 bp
   Segment 3:  LTR sequences  
   364 .. 632  =  269 bp
hybrid promoter consisting of the SV40 early
promoter plus part of the long terminal repeat of
human T-lymphotrophic virus 1
SRα promoter
18 .. 632  =  615 bp
3 segments
hybrid promoter consisting of the SV40 early
promoter plus part of the long terminal repeat of
human T-lymphotrophic virus 1
ori
1641 .. 2229  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1641 .. 2229  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
stuffer
812 .. 1194  =  383 bp
stuffer
812 .. 1194  =  383 bp
SV40 poly(A) signal
1297 .. 1431  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1297 .. 1431  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3261 .. 3365  =  105 bp
AmpR promoter
3261 .. 3365  =  105 bp
SV40 intron
662 .. 755  =  94 bp
modified SV40 intron with splice donor and acceptor
sites
SV40 intron
662 .. 755  =  94 bp
modified SV40 intron with splice donor and acceptor
sites
SV40 ori
198 .. 333  =  136 bp
SV40 origin of replication
SV40 ori
198 .. 333  =  136 bp
SV40 origin of replication
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