pOTB7-3

cDNA cloning vector with an f1 origin derived from pOTB7.

Sequence Author: I.M.A.G.E. Consortium

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SalI - SgrDI (2812) ScaI (2775) BtgI - NcoI - StyI (2659) MscI (2625) PasI - PflMI * (2590) BsmBI (2582) BpmI (2480) AclI (2445) BsrDI (2377) BspEI (2354) Bpu10I (2131) BspDI - ClaI (2080) BssSI (1887) BciVI (1862) BaeGI - Bme1580I (1750) ApaLI (1746) AlwNI (1651) AcuI (1518) AccI (2813) M13 fwd Bsu36I (141) StuI * (154) BglI (160) XhoI (185) BstXI (204) NspI (216) BspQI - EarI - SapI (329) BtsI (432) SP6 promoter NsiI (610) HindIII (613) Acc65I (619) KpnI (623) Eco53kI (627) SacI (629) BstXI (658) EcoRV (674) BsaBI (676) BstXI (684) XhoI (695) PI-SceI (717) M13 rev BglII (788) AscI - BssHII (797) BsaI (819) HpaI (855) BfuAI - BspMI (866) AanI - PsiI (986) BsaAI - DraIII (1114) NgoMIV (1215) NaeI (1217) pOTB7-3 2816 bp
SalI  (2812)
1 site
G T C G A C C A G C T G
SgrDI  (2812)
1 site
C G T C G A C G G C A G C T G C
ScaI  (2775)
1 site
A G T A C T T C A T G A
BtgI  (2659)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (2659)
1 site
C C A T G G G G T A C C
StyI  (2659)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (2625)
1 site
T G G C C A A C C G G T
PasI  (2590)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (2590)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BsmBI  (2582)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BpmI  (2480)
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.
AclI  (2445)
1 site
A A C G T T T T G C A A
BsrDI  (2377)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
BspEI  (2354)
1 site
T C C G G A A G G C C T
Bpu10I  (2131)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BspDI  (2080)
1 site
A T C G A T T A G C T A
ClaI  (2080)
1 site
A T C G A T T A G C T A
BssSI  (1887)
1 site
C A C G A G G T G C T C
BciVI  (1862)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BaeGI  (1750)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (1750)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
ApaLI  (1746)
1 site
G T G C A C C A C G T G
AlwNI  (1651)
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.
AcuI  (1518)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AccI  (2813)
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.
Bsu36I  (141)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
StuI  (154)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BglI  (160)
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.
XhoI  (185)
2 sites
C T C G A G G A G C T C
BstXI  (204)
3 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.
NspI  (216)
1 site
R C A T G Y Y G T A C R
BspQI  (329)
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.
EarI  (329)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
SapI  (329)
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.
BtsI  (432)
1 site
G C A G T G N N C G T C A C
NsiI  (610)
1 site
A T G C A T T A C G T A
HindIII  (613)
1 site
A A G C T T T T C G A A
Acc65I  (619)
1 site
G G T A C C C C A T G G
KpnI  (623)
1 site
G G T A C C C C A T G G
Eco53kI  (627)
1 site
G A G C T C C T C G A G
SacI  (629)
1 site
G A G C T C C T C G A G
BstXI  (658)
3 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.
EcoRV  (674)
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.
BsaBI  (676)
1 site
G A T N N N N A T C C T A N N N N T A G
BstXI  (684)
3 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.
XhoI  (695)
2 sites
C T C G A G G A G C T C
PI-SceI  (717)
1 site
A T C T A T G T C G G G T G C G G A G A A A G A G G T A A T G A A A T G G T A G A T A C A G C C C A C G C C T C T T T C T C C A T T A C T T T A C C

PI-SceI  is a homing endonuclease that can recognize a variety of similar recognition sequences.
BglII  (788)
1 site
A G A T C T T C T A G A
AscI  (797)
1 site
G G C G C G C C C C G C G C G G
BssHII  (797)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BsaI  (819)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
HpaI  (855)
1 site
G T T A A C C