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Plasmid Files

pMT/V5-His-TOPO® (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products and inducible expression in Drosophila cells.

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pMT V5-His-TOPO (linearized) Sequence and MappMT V5-His-TOPO (linearized).dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 ZraI (2609) SspI (2493) ScaI (2169) NmeAIII (1837) BpmI (1759) AhdI (1689) AlwNI (1212) PspFI (1104) BseYI (1100) AatII (2611) PfoI (2721) BstAPI (2860) KasI (2910) NarI (2911) SfoI (2912) PluTI (2914) BsgI (3315) BmgBI (3337) MscI (3446) XbaI * (3514) Acc65I (3524) KpnI (3528) SpeI (3531) BstXI (3544) End (3558) Start (1) EcoRV (18) BstXI (28) NotI (33) AvaI - BsoBI - PaeR7I - PspXI - XhoI (39) BmeT110I (40) XbaI (45) PspOMI (51) ApaI - BanII - BtgI (55) SacII (58) BstBI (62) BseRI (79) MluI (106) AgeI (112) PmeI (142) AanI - PsiI (291) HpaI (311) MfeI (320) SalI (419) AccI (420) SbfI (429) BfuAI - BspMI (432) BspQI - SapI (680) PciI (796) pMT/V5-His-TOPO® 3557 bp
ZraI  (2609)
1 site
G A C G T C C T G C A G
SspI  (2493)
1 site
A A T A T T T T A T A A
ScaI  (2169)
1 site
A G T A C T T C A T G A
NmeAIII  (1837)
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).
BpmI  (1759)
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  (1689)
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  (1212)
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  (1104)
1 site
C C C A G C G G G T C G
BseYI  (1100)
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.
AatII  (2611)
1 site
G A C G T C C T G C A G
PfoI  (2721)
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.
BstAPI  (2860)
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.
KasI  (2910)
1 site
G G C G C C C C G C G G
NarI  (2911)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (2912)
1 site
G G C G C C C C G C G G
PluTI  (2914)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
BsgI  (3315)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

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).
BmgBI  (3337)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
MscI  (3446)
1 site
T G G C C A A C C G G T
XbaI  (3514)
2 sites
T C T A G A A G A T C T
* Blocked by Dam methylation.
Acc65I  (3524)
1 site
G G T A C C C C A T G G
KpnI  (3528)
1 site
G G T A C C C C A T G G
SpeI  (3531)
1 site
A C T A G T T G A T C A
BstXI  (3544)
2 sites
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.
End  (3558)
0 sites
Start  (1)
0 sites
EcoRV  (18)
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.
BstXI  (28)
2 sites
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.
NotI  (33)
1 site
G C G G C C G C C G C C G G C G
AvaI  (39)
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  (39)
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.
PaeR7I  (39)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (39)
1 site
V C T C G A G B B G A G C T C V
XhoI  (39)
1 site
C T C G A G G A G C T C
BmeT110I  (40)
1 site
C Y C G R G G R G C Y C
XbaI  (45)
2 sites
T C T A G A A G A T C T
PspOMI  (51)
1 site
G G G C C C C C C G G G
ApaI  (55)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (55)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BtgI  (55)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (58)
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.
BstBI  (62)
1 site
T T C G A A A A G C T T
BseRI  (79)
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.
MluI  (106)
1 site
A C G C G T T G C G C A
AgeI  (112)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
PmeI  (142)
1 site
G T T T A A A C C A A A T T T G
AanI  (291)
1 site
T T A T A A A A T A T T
PsiI  (291)
1 site
T T A T A A A A T A T T
HpaI  (311)
1 site
G T T A A C C A A T T G
MfeI  (320)
1 site
C A A T T G G T T A A C
SalI  (419)
1 site
G T C G A C C A G C T G
AccI  (420)
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.
SbfI  (429)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (432)
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  (432)
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.
BspQI  (680)
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  (680)
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.
PciI  (796)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
1616 .. 2476  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1616 .. 2407  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1616 .. 2476  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2408 .. 2476  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1616 .. 2476  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
857 .. 1445  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
857 .. 1445  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
MT promoter
3082 .. 3508  =  427 bp
Drosophila metallothionein promoter
MT promoter
3082 .. 3508  =  427 bp
Drosophila metallothionein promoter
SV40 poly(A) signal
177 .. 311  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
177 .. 311  =  135 bp
SV40 polyadenylation signal
AmpR promoter
2477 .. 2581  =  105 bp
AmpR promoter
2477 .. 2581  =  105 bp
V5 tag
67 .. 108  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
67 .. 108  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
118 .. 135  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
118 .. 135  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
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