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Plasmid Files

pETBlue™-2

Vector for blue/white screening and for inducible bacterial expression of tagged proteins.

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pETBlue-2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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AgeI (3599) NspI (3267) PciI (3263) PspFI (2963) BseYI (2959) AlwNI (2854) BspHI (2543) XmnI (2338) ScaI (2219) NmeAIII (1887) XbaI (47) BglII (223) NheI (235) BmtI (239) BtgI - NcoI (276) EcoRV (285) TspMI - XmaI (288) SmaI (290) Eco53kI (295) SacI (297) BamHI (301) EcoRI (307) BsrGI (313) AscI - BssHII (320) PstI - SbfI (330) ZraI (334) SalI - SgrDI (335) AatII (336) HincII (337) Acc65I (341) KpnI (345) BspDI - ClaI (348) MluI (353) BstBI (359) HindIII (362) EagI - NotI (369) PvuII (379) BstZ17I (384) PmlI (389) PaeR7I - XhoI (431) PacI (464) AvrII (597) BlpI (1001) BspEI (1078) BtgZI (1321) DraIII (1329) PsiI (1454) SspI (1534) AhdI (1739) pETBlue™-2 3653 bp
AgeI  (3599)
1 site
A C C G G T T G G C C A
NspI  (3267)
1 site
R C A T G Y Y G T A C R
PciI  (3263)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2963)
1 site
C C C A G C G G G T C G
BseYI  (2959)
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.
AlwNI  (2854)
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.
BspHI  (2543)
1 site
T C A T G A A G T A C T
XmnI  (2338)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2219)
1 site
A G T A C T T C A T G A
NmeAIII  (1887)
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).
XbaI  (47)
1 site
T C T A G A A G A T C T
BglII  (223)
1 site
A G A T C T T C T A G A
NheI  (235)
1 site
G C T A G C C G A T C G
BmtI  (239)
1 site
G C T A G C C G A T C G
BtgI  (276)
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  (276)
1 site
C C A T G G G G T A C C
EcoRV  (285)
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.
TspMI  (288)
1 site
C C C G G G G G G C C C
XmaI  (288)
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  (290)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Eco53kI  (295)
1 site
G A G C T C C T C G A G
SacI  (297)
1 site
G A G C T C C T C G A G
BamHI  (301)
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.
EcoRI  (307)
1 site
G A A T T C C T T A A G
BsrGI  (313)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AscI  (320)
1 site
G G C G C G C C C C G C G C G G
BssHII  (320)
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.
PstI  (330)
1 site
C T G C A G G A C G T C
SbfI  (330)
1 site
C C T G C A G G G G A C G T C C
ZraI  (334)
1 site
G A C G T C C T G C A G
SalI  (335)
1 site
G T C G A C C A G C T G
SgrDI  (335)
1 site
C G T C G A C G G C A G C T G C
AatII  (336)
1 site
G A C G T C C T G C A G
HincII  (337)
1 site
G T Y R A C C A R Y T G
Acc65I  (341)
1 site
G G T A C C C C A T G G