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: DownloadOpen Sequence Author: Thermo Fisher (Invitrogen)
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI (3830) 1 site
GGCGCCCCGCGG
NarI (3829) 1 site
GGCGCCCCGCGG
* Blocked by Dcm methylation.Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI (3828) 1 site
GGCGCCCCGCGG
EcoRV (3639) 1 site
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
ApaI (3400) 1 site
GGGCCCCCCGGG
ApaI can be used between 25°C and 37°C.
BanII (3400) 1 site
GRGCYCCYCGRG
Sticky ends from different BanII sites may not be compatible.
PspOMI (3396) 1 site
GGGCCCCCCGGG
BstEII (3370) 1 site
GGTNACCCCANTGG
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
TGATCAACTAGT
* Blocked by Dam methylation.BclI is typically used at 50-55°C, but is 50% active at 37°C.
MluI (3189) 1 site
ACGCGTTGCGCA
PflMI (2771) 1 site
CCANNNNNTGGGGTNNNNNACC
Sticky ends from different PflMI sites may not be compatible.
NsiI (2752) 1 site
ATGCATTACGTA
SphI (2750) 1 site
GCATGCCGTACG
MauBI (2728) 1 site
CGCGCGCGGCGCGCGC
PfoI (2660) 1 site
TCCNGGAAGGNCCT
Sticky ends from different PfoI sites may not be compatible.
PflFI (2561) 1 site
GACNNNGTCCTGNNNCAG
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
GACNNNGTCCTGNNNCAG
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
GTATACCATATG
NdeI (2484) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BspQI (2422) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different BspQI sites may not be compatible.
SapI (2422) 1 site
GCTCTTCNCGAGAAGNNNN
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.