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pTrcHis2-TOPO® (linearized)

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

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pTrcHis2-TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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PluTI (3832) SfoI (3830) NarI * (3829) KasI (3828) EcoRV (3639) ApaI - BanII (3400) PspOMI (3396) BstEII (3370) BclI * (3203) MluI (3189) PflMI (2771) NsiI (2752) SphI (2750) MauBI (2728) PfoI (2660) PflFI - Tth111I (2561) BstZ17I (2536) NdeI (2484) BspQI - SapI (2422) PciI (2305) AlwNI (1896) BfuAI - BspMI (4032) BtgI - NcoI - StyI (4371) End (4381) Start (1) EcoRI (7) BstBI (11) HindIII (14) SnaBI (21) SalI (64) PmeI (94) ScaI (936) TsoI (1019) PvuI (1048) FspI (1194) BglI (1299) AhdI (1417) pTrcHis2-TOPO® 4380 bp
PluTI  (3832)
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.
SfoI  (3830)
1 site
G G C G C C C C G C G G
NarI  (3829)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (3828)
1 site
G G C G C C C C G C G G
EcoRV  (3639)
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.
ApaI  (3400)
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  (3400)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (3396)
1 site
G G G C C C C C C G G G
BstEII  (3370)
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.
BclI  (3203)
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.
MluI  (3189)
1 site
A C G C G T T G C G C A
PflMI  (2771)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
NsiI  (2752)
1 site
A T G C A T T A C G T A
SphI  (2750)
1 site
G C A T G C C G T A C G
MauBI  (2728)
1 site
C G C G C G C G G C G C G C G C
PfoI  (2660)
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.
PflFI  (2561)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2561)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BstZ17I  (2536)
1 site
G T A T A C C A T A T G
NdeI  (2484)
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.
BspQI  (2422)
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  (2422)
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  (2305)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (1896)
1 site
C A G N N N