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Plasmid Files

pExpress-1

Mammalian expression vector for constructing cDNA libraries.

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pExpress-1 Sequence and MappExpress-1.dna
Map and Sequence File   
Sequence Author:  I.M.A.G.E. Consortium
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 NaeI (3988) NgoMIV (3986) DraIII (3885) XmnI (3341) ScaI (3222) NmeAIII (2890) BsaI (2803) AhdI (2742) AlwNI (2265) PspFI (2157) BspDI * - ClaI * (123) HincII - HpaI (259) MfeI (268) BsaBI * (360) BclI * (361) NheI (637) BmtI (641) BseRI (706) AbsI - PaeR7I - PspXI - XhoI (718) PacI (729) EagI - NotI (734) EcoRV (743) TspMI - XmaI (746) SmaI (748) EcoRI (751) RsrII (758) SP6 promoter StuI (793) M13 rev SacII (840) AvrII (843) BbsI (872) BsmBI (917) Eco53kI (944) SacI (946) NcoI (1146) SnaBI (1170) NsiI (1457) BstAPI (1673) BspQI - SapI (1733) AflIII - PciI (1849) BseYI (2153) pExpress-1 4117 bp
NaeI  (3988)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (3986)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
DraIII  (3885)
1 site
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.
XmnI  (3341)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3222)
1 site
A G T A C T T C A T G A
NmeAIII  (2890)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaI  (2803)
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  (2742)
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.
AlwNI  (2265)
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  (2157)
1 site
C C C A G C G G G T C G
BspDI  (123)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (123)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
HincII  (259)
1 site
G T Y R A C C A R Y T G
HpaI  (259)
1 site
G T T A A C C A A T T G
MfeI  (268)
1 site
C A A T T G G T T A A C
BsaBI  (360)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BclI  (361)
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.
NheI  (637)
1 site
G C T A G C C G A T C G
BmtI  (641)
1 site
G C T A G C C G A T C G
BseRI  (706)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
AbsI  (718)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (718)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (718)
1 site
V C T C G A G B B G A G C T C V
XhoI  (718)
1 site
C T C G A G G A G C T C
PacI  (729)
1 site
T T A A T T A A A A T T A A T T
EagI  (734)
1 site
C G G C C G G C C G G C
NotI  (734)
1 site
G C G G C C G C C G C C G G C G
EcoRV  (743)
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.
TspMI  (746)
1 site
C C C G G G G G G C C C
XmaI  (746)
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  (748)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (751)
1 site
G A A T T C C T T A A G
RsrII  (758)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
StuI  (793)
1 site
A G G C C T T C C G G A
SacII  (840)
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.
AvrII  (843)
1 site
C C T A G G G G A T C C
BbsI  (872)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsmBI  (917)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
Eco53kI  (944)
1 site
G A G C T C C T C G A G
SacI  (946)
1 site
G A G C T C C T C G A G
NcoI  (1146)
1 site
C C A T G G G G T A C C
SnaBI  (1170)
1 site
T A C G T A A T G C A T
NsiI  (1457)
1 site
A T G C A T T A C G T A
BstAPI  (1673)
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.
BspQI  (1733)
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  (1733)
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.
AflIII  (1849)
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  (1849)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (2153)
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.
AmpR
2669 .. 3529  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2669 .. 3460  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2669 .. 3529  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3461 .. 3529  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2669 .. 3529  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1910 .. 2498  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1910 .. 2498  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3661 .. 4116  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3661 .. 4116  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
1147 .. 1450  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1147 .. 1450  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
943 .. 1146  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
943 .. 1146  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
125 .. 259  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
125 .. 259  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3530 .. 3634  =  105 bp
AmpR promoter
3530 .. 3634  =  105 bp
MCS
718 .. 763  =  46 bp
multiple cloning site
MCS
718 .. 763  =  46 bp
multiple cloning site
T7 promoter
691 .. 709  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
691 .. 709  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
772 .. 790  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
772 .. 790  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 fwd
664 .. 680  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
664 .. 680  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
801 .. 817  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
801 .. 817  =  17 bp
common sequencing primer, one of multiple similar
variants
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