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Plasmid Files

pEXP5-CT/TOPO® (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning and inducible bacterial expression of a C-terminally 6xHis-tagged protein.

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pEXP5-CT TOPO (linearized) Sequence and MappEXP5-CT TOPO (linearized).dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 BmeT110I (2582) AvaI - BsoBI (2581) MscI * (2564) BtgI (2559) FspAI (2554) BsgI * (2377) BtgZI (2347) BstZ17I (2262) AccI (2261) BspQI - SapI (2149) NspI (2036) AflIII - PciI (2032) DrdI (1930) PspFI (1732) BseYI (1728) AlwNI (1623) BanI (1191) BglII (2585) XbaI (2644) End (2685) Start (1) PmeI (31) BlpI (63) StyI (85) ZraI (223) AatII (225) SspI (339) XmnI (544) HincII (602) ScaI (663) PvuI (775) NmeAIII (997) BglI (1026) BsrFI (1059) BpmI (1075) BmrI (1104) AhdI (1144) pEXP5-CT/TOPO® 2684 bp
BmeT110I  (2582)
1 site
C Y C G R G G R G C Y C
AvaI  (2581)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2581)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
MscI  (2564)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BtgI  (2559)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
FspAI  (2554)
1 site
R T G C G C A Y Y A C G C G T R
BsgI  (2377)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BtgZI  (2347)
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.
BstZ17I  (2262)
1 site
G T A T A C C A T A T G
AccI  (2261)
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.
BspQI  (2149)
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  (2149)
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.
NspI  (2036)
1 site
R C A T G Y Y G T A C R
AflIII  (2032)
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  (2032)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (1930)
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.
PspFI  (1732)
1 site
C C C A G C G G G T C G
BseYI  (1728)
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.
AlwNI  (1623)
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.
BanI  (1191)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
BglII  (2585)
1 site
A G A T C T T C T A G A
XbaI  (2644)
1 site
T C T A G A A G A T C T
End  (2685)
0 sites
Start  (1)
0 sites
PmeI  (31)
1 site
G T T T A A A C C A A A T T T G
BlpI  (63)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
StyI  (85)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
ZraI  (223)
1 site
G A C G T C C T G C A G
AatII  (225)
1 site
G A C G T C C T G C A G
SspI  (339)
1 site
A A T A T T T T A T A A
XmnI  (544)
1 site
G A A N N N N T T C C T T N N N N A A G
HincII  (602)
1 site
G T Y R A C C A R Y T G
ScaI  (663)
1 site
A G T A C T T C A T G A
PvuI  (775)
1 site
C G A T C G G C T A G C
NmeAIII  (997)
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).
BglI  (1026)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BsrFI  (1059)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI  (1075)
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.
BmrI  (1104)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
AhdI  (1144)
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.
AmpR
357 .. 1217  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   357 .. 425  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
357 .. 1217  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   426 .. 1217  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
357 .. 1217  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1388 .. 1976  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
1388 .. 1976  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
252 .. 356  =  105 bp
AmpR promoter
252 .. 356  =  105 bp
T7 terminator
74 .. 121  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
74 .. 121  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 promoter
2605 .. 2623  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2605 .. 2623  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
7 .. 24  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
7 .. 24  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
2673 .. 2678  =  6 bp
ribosome binding site
RBS
2673 .. 2678  =  6 bp
ribosome binding site
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