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Plasmid Files

pSI

Mammalian cell expression vector with the SV40 promoter.

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pSI Sequence and MappSI.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (3627) AlwNI (3206) AhdI (2727) BpmI (2658) AseI (2552) XmnI (2127) AatII (1808) PvuII (74) SexAI * (165) SfiI (351) BseRI (394) StuI (397) AvrII (398) HindIII (414) PstI (456) BfuAI - BspMI (470) BbsI (554) NheI (678) BmtI (682) PaeR7I - XhoI (684) EcoRI (689) AflIII - MluI (695) XbaI (707) SalI (713) AccI (714) TspMI - XmaI (718) SmaI (720) EagI - NotI (724) HpaI (867) MfeI (876) BspDI - ClaI (962) BsaBI * (968) BamHI (969) NgoMIV (1173) NaeI (1175) BanII (1207) BtgZI (1273) BsaAI (1278) DraIII (1281) NdeI (1553) PfoI (1688) EcoO109I (1747) ZraI (1806) pSI 3632 bp
BglII  (3627)
1 site
A G A T C T T C T A G A
AlwNI  (3206)
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.
AhdI  (2727)
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.
BpmI  (2658)
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.
AseI  (2552)
1 site
A T T A A T T A A T T A
XmnI  (2127)
1 site
G A A N N N N T T C C T T N N N N A A G
AatII  (1808)
1 site
G A C G T C C T G C A G
PvuII  (74)
1 site
C A G C T G G T C G A C
SexAI  (165)
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.
SfiI  (351)
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.
BseRI  (394)
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.
StuI  (397)
1 site
A G G C C T T C C G G A
AvrII  (398)
1 site
C C T A G G G G A T C C
HindIII  (414)
1 site
A A G C T T T T C G A A
PstI  (456)
1 site
C T G C A G G A C G T C
BfuAI  (470)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (470)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BbsI  (554)
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.
NheI  (678)
1 site
G C T A G C C G A T C G
BmtI  (682)
1 site
G C T A G C C G A T C G
PaeR7I  (684)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (684)
1 site
C T C G A G G A G C T C
EcoRI  (689)
1 site
G A A T T C C T T A A G
AflIII  (695)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (695)
1 site
A C G C G T T G C G C A
XbaI  (707)
1 site
T C T A G A A G A T C T
SalI  (713)
1 site
G T C G A C C A G C T G
AccI  (714)
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.
TspMI  (718)
1 site
C C C G G G G G G C C C
XmaI  (718)
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  (720)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EagI  (724)
1 site
C G G C C G G C C G G C
NotI  (724)
1 site
G C G G C C G C C G C C G G C G
HpaI  (867)
1 site
G T T A A C C A A T T G
MfeI  (876)
1 site
C A A T T G G T T A A C
BspDI  (962)
1 site
A T C G A T T A G C T A
ClaI  (962)
1 site
A T C G A T T A G C T A
BsaBI  (968)
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.
BamHI  (969)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
NgoMIV  (1173)
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.
NaeI  (1175)
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.
BanII  (1207)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BtgZI  (1273)
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.
BsaAI  (1278)
1 site
Y A C G T R R T G C A Y
DraIII  (1281)
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.
NdeI  (1553)
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  (1688)
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.
EcoO109I  (1747)
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.
ZraI  (1806)
1 site
G A C G T C C T G C A G
AmpR
1940 .. 2800  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1940 .. 2008  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1940 .. 2800  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2009 .. 2800  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1940 .. 2800  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2971 .. 3559  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2971 .. 3559  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1048 .. 1503  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1048 .. 1503  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
56 .. 413  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
56 .. 413  =  358 bp
SV40 enhancer and early promoter
chimeric intron
483 .. 615  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
483 .. 615  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
746 .. 867  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
746 .. 867  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1835 .. 1939  =  105 bp
AmpR promoter
1835 .. 1939  =  105 bp
MCS
678 .. 730  =  53 bp
multiple cloning site
MCS
678 .. 730  =  53 bp
multiple cloning site
SV40 ori
264 .. 399  =  136 bp
SV40 origin of replication
SV40 ori
264 .. 399  =  136 bp
SV40 origin of replication
T7 promoter
660 .. 678  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
660 .. 678  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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