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Plasmid Files

pSEAP2-Basic

Promoterless mammalian vector encoding secreted alkaline phosphatase, for cloning promoter and enhancer elements.

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pSEAP2-Basic Sequence and MappSEAP2-Basic.dna
Map and Sequence File   
Sequence Author:  Clontech
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 AarI - BfuAI - BspMI (4640) poly(A) signal NotI (4510) BsaAI (4161) XmnI (3611) AseI (3184) AhdI (3012) Acc65I (1) KpnI (5) MluI (15) NheI (21) BmtI (25) SrfI (28) PaeR7I - XhoI (32) BglII (36) HindIII (53) BstBI (58) NruI (65) EcoRI (67) Bsu36I (107) PstI (210) PvuII (281) MscI * (338) NdeI (355) BstXI (458) BstEII (565) AleI (578) AvrII (727) SacII (1042) BsmBI (1112) XcmI (1255) StuI (1289) BssHII - MauBI (1451) SexAI * (1470) XbaI (1601) FseI (1620) HpaI (1761) MfeI (1770) BsaBI * (1862) SalI (1869) AfeI (1995) BspQI - SapI (2003) pSEAP2-Basic 4677 bp
AarI  (4640)
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.
BfuAI  (4640)
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  (4640)
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.
NotI  (4510)
1 site
G C G G C C G C C G C C G G C G
BsaAI  (4161)
1 site
Y A C G T R R T G C A Y
XmnI  (3611)
1 site
G A A N N N N T T C C T T N N N N A A G
AseI  (3184)
1 site
A T T A A T T A A T T A
AhdI  (3012)
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.
Acc65I  (1)
1 site
G G T A C C C C A T G G
KpnI  (5)
1 site
G G T A C C C C A T G G
MluI  (15)
1 site
A C G C G T T G C G C A
NheI  (21)
1 site
G C T A G C C G A T C G
BmtI  (25)
1 site
G C T A G C C G A T C G
SrfI  (28)
1 site
G C C C G G G C C G G G C C C G
PaeR7I  (32)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (32)
1 site
C T C G A G G A G C T C
BglII  (36)
1 site
A G A T C T T C T A G A
HindIII  (53)
1 site
A A G C T T T T C G A A
BstBI  (58)
1 site
T T C G A A A A G C T T
NruI  (65)
1 site
T C G C G A A G C G C T
EcoRI  (67)
1 site
G A A T T C C T T A A G
Bsu36I  (107)
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.
PstI  (210)
1 site
C T G C A G G A C G T C
PvuII  (281)
1 site
C A G C T G G T C G A C
MscI  (338)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
NdeI  (355)
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.
BstXI  (458)
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.
BstEII  (565)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
AleI  (578)
1 site
C A C N N N N G T G G T G N N N N C A C
AvrII  (727)
1 site
C C T A G G G G A T C C
SacII  (1042)
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.
BsmBI  (1112)
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.
XcmI  (1255)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
StuI  (1289)
1 site
A G G C C T T C C G G A
BssHII  (1451)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
MauBI  (1451)
1 site
C G C G C G C G G C G C G C G C
SexAI  (1470)
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.
XbaI  (1601)
1 site
T C T A G A A G A T C T
FseI  (1620)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
HpaI  (1761)
1 site
G T T A A C C A A T T G
MfeI  (1770)
1 site
C A A T T G G T T A A C
BsaBI  (1862)
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.
SalI  (1869)
1 site
G T C G A C C A G C T G
AfeI  (1995)
1 site
A G C G C T T C G C G A
BspQI  (2003)
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  (2003)
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.
SEAP
80 .. 1582  =  1503 bp
501 amino acids  =  54.6 kDa
Product: secreted alkaline phosphatase from human
placenta
The natural enzyme has a C-terminal
transmembrane sequence that is replaced by a GPI
anchor.
SEAP
80 .. 1582  =  1503 bp
501 amino acids  =  54.6 kDa
Product: secreted alkaline phosphatase from human
placenta
The natural enzyme has a C-terminal
transmembrane sequence that is replaced by a GPI
anchor.
AmpR
2939 .. 3799  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2939 .. 3730  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2939 .. 3799  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3731 .. 3799  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2939 .. 3799  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2180 .. 2768  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2180 .. 2768  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3931 .. 4386  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3931 .. 4386  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 poly(A) signal
1640 .. 1761  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1640 .. 1761  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3800 .. 3904  =  105 bp
AmpR promoter
3800 .. 3904  =  105 bp
pause site
4579 .. 4670  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4579 .. 4670  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
MCS
1 .. 72  =  72 bp
multiple cloning site
MCS
1 .. 72  =  72 bp
multiple cloning site
poly(A) signal
4517 .. 4565  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4517 .. 4565  =  49 bp
synthetic polyadenylation signal
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