pME18S-FL3

Mammalian vector for directional cloning and transient expression of a cDNA.
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HomePlasmidsI.M.A.G.E. Consortium PlasmidspME18S-FL3
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
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
2 segments
   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
2 segments
   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
3 segments
   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
3 segments
   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
3 segments
   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
ORF:  2530 .. 2796  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  869 .. 1228  =  360 bp
ORF:  119 amino acids  =  13.4 kDa
ORF:  1280 .. 1540  =  261 bp
ORF:  86 amino acids  =  9.7 kDa
ORF:  2400 .. 3260  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

Lafmid BA
pcDNAII
pCR2.1-TOPO (linearized)
pExpress-1
pAD-GAL4
pcDNA3.1(+)
pCR3.1
pME18S-FL3
pAMP1
pcDNA3.1(-)
pCR3.1 (linearized)
pOTB7
pAMP10
pcDNA3.1 Zeo(+)
pCR4-TOPO
pOTB7-3
pBK-CMV
pCMV SPORT
pCR4-TOPO (linearized)
pOTB7a
pBluescribe (modified)
pCMV SPORT2
pCS105
pPCR-Script Amp SK(+)
pBluescript II SK(+)
pCMV SPORT4
pCS107
pRKW2
pBluescript KS(+)
pCMV SPORT6
pCS108
pSPORT 1
pBluescript SK(+)
pCMV SPORT6.1
pCS2+
pSPORT 2
pBluescript SK(-)
pCMV SPORT6ccdB
pCS22+
pT7T3D-PacI
pBluescript-FL
pCR-Blunt II-TOPO
pCS2G
pUC19
pBluescript II KS(+)
pCR-Blunt II-TOPO (linearized)
pDNR-Dual
pYX-Asc
pBluescriptR
pCR-XL-TOPO
pDNR-LIB
pZErO-2
pcDNA3
pCR-XL-TOPO (linearized)
pDONR201
pZL1
pcDNAI
pCR2.1-TOPO
pDONR223

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