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pTWV228

Low copy number bacterial vector for cloning sequences that cause toxicity.

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pTWV228.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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ZraI (3972) ScaI (3532) AseI (3224) NmeAIII (3200) BsaI (3113) AhdI (3052) AlwNI (2575) PspFI (2467) BseYI (2463) AflIII - PciI (2159) BspQI - SapI (2043) BstAPI (1983) AatII (3974) PsiI (283) DraIII (411) NgoMIV (512) NaeI (514) KasI (711) NarI (712) SfoI (713) PluTI (715) HindIII (875) BfuAI - BspMI (880) SphI (885) PstI - SbfI (891) SalI (893) HincII (895) XbaI (899) BamHI (905) TspMI - XmaI (910) SmaI (912) Acc65I (914) KpnI (918) Eco53kI (922) SacI (924) EcoRI (926) MscI * (1132) BtgI (1133) FspAI (1142) Bpu10I (1267) BbsI (1279) BsgI * (1321) BspEI * (1350) BsaBI * (1358) AfeI (1415) PflFI - Tth111I (1906) BstZ17I (1932) NdeI (1982) pTWV228 4039 bp
ZraI  (3972)
1 site
G A C G T C C T G C A G
ScaI  (3532)
1 site
A G T A C T T C A T G A
AseI  (3224)
1 site
A T T A A T T A A T T A
NmeAIII  (3200)
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).
BsaI  (3113)
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.
AhdI  (3052)
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  (2575)
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  (2467)
1 site
C C C A G C G G G T C G
BseYI  (2463)
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.
AflIII  (2159)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2159)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2043)
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  (2043)
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.
BstAPI  (1983)
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.
AatII  (3974)
1 site
G A C G T C C T G C A G
PsiI  (283)
1 site
T T A T A A A A T A T T
DraIII  (411)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
NgoMIV  (512)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (514)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
KasI  (711)
1 site
G G C G C C C C G C G G
NarI  (712)
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  (713)
1 site
G G C G C C C C G C G G
PluTI  (715)
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.
HindIII  (875)
1 site
A A G C T T T T C G A A
BfuAI  (880)
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  (880)
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.
SphI  (885)
1 site
G C A T G C C G T A C G
PstI  (891)
1 site
C T G C A G G A C G T C
SbfI  (891)
1 site
C C T G C A G G G G A C G T C C
SalI  (893)
1 site
G T C G A C C A G C T G
HincII  (895)
1 site
G T Y R A C C A R Y T G
XbaI  (899)
1 site
T C T A G A A G A T C T
BamHI  (905)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
TspMI  (910)
1 site
C C C G G G G G G C C C
XmaI  (910)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (912)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (914)
1 site
G G T A C C C C A T G G
KpnI  (918)
1 site
G G T A C C C C A T G G
Eco53kI  (922)
1 site
G A G C T C C T C G A G
SacI  (924)
1 site
G A G C T C C T C G A G
EcoRI  (926)
1 site
G A A T T C C T T A A G
MscI  (1132)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BtgI  (1133)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
FspAI  (1142)
1 site
R T G C G C A Y Y A C G C G T R
Bpu10I  (1267)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
BbsI  (1279)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsgI  (1321)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
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).
BspEI  (1350)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
BsaBI  (1358)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AfeI  (1415)
1 site