pOSIP-TO

Prokaryotic one-step cloning and chromosomal integration vector encoding a tetracycline resistance marker and the phage 186 integrase.
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EcoRI (1) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator BsgI (6104) PflFI - Tth111I (6090) PfoI (5987) λ tL3 terminator BseRI (5742) SnaBI (4883) SalI (4798) StuI (4659) ScaI (4265) BglI (4222) PmlI (3988) EcoRV (3546) Acc65I (13) KpnI (17) BamHI (22) BssHII (70) SrfI (225) BsaI (363) BglII (469) BciVI (923) SpeI (1126) MCS 2 PstI (1144) rrnB T2 terminator BstBI (1234) PacI (1340) PspOMI (1349) ApaI (1353) ZraI (1379) AatII (1381) AvrII (1418) EarI (1442) BsmI (1718) PshAI (2090) AfeI (2473) NruI (2921) FspAI (3213) DraIII (3360) BspDI - ClaI (3366) pOSIP-TO 6919 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
BsgI  (6104)
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).
PflFI  (6090)
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  (6090)
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.
PfoI  (5987)
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.
BseRI  (5742)
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.
SnaBI  (4883)
1 site
T A C G T A A T G C A T
SalI  (4798)
1 site
G T C G A C C A G C T G
StuI  (4659)
1 site
A G G C C T