pEZ BAC

BAC vector with inducible copy number, suitable for cloning toxic inserts.

Sequence Author: Lucigen

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SacII (7159) FseI (7155) BciVI - MreI (7144) XcmI (6954) XbaI (6444) EcoNI (6170) SmaI (6147) TspMI - XmaI (6145) FspAI (5888) BsrGI (5856) SbfI (5619) BlpI (5158) AfeI (5074) MfeI (4649) NdeI (4633) BstXI (4559) KpnI (4433) Acc65I (4429) PflFI - Tth111I (4368) BseRI (3942) BclI * (3838) T3Te terminator AscI (120) NotI (148) BEZ-F1 (238 .. 261) BsrBI (278) lac operator HindIII (355) SphI (365) HpaI (379) BamHI (386) PmlI (398) EcoRI (403) BEZ-R1 (488 .. 511) KasI (590) NarI (591) SfoI (592) BbeI (594) NotI (620) PmeI (669) tonB terminator EcoO109I - PpuMI (983) DraIII (1024) ScaI (1918) BstAPI (1998) BstEII (2031) SexAI * (2035) PciI (2154) AhdI (2155) PspOMI (2668) ApaI - BanII (2672) AccI (2744) PfoI * (2884) SpeI (2914) AleI (3097) pEZ™ BAC 7260 bp
SacII  (7159)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
FseI  (7155)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BciVI  (7144)
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.
MreI  (7144)
1 site
C G C C G G C G G C G G C C G C
XcmI  (6954)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
XbaI  (6444)
1 site
T C T A G A A G A T C T
EcoNI  (6170)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SmaI  (6147)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (6145)
1 site
C C C G G G G G G C C C
XmaI  (6145)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
FspAI  (5888)
1 site
R T G C G C A Y Y A C G C G T R
BsrGI  (5856)
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.
SbfI  (5619)
1 site
C C T G C A G G G G A C G T C C
BlpI  (5158)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
AfeI  (5074)
1 site
A G C G C T T C G C G A
MfeI  (4649)
1 site
C A A T T G G T T A A C
NdeI  (4633)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BstXI  (4559)
1 site
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.
KpnI  (4433)
1 site
G G T A C C C C A T G G
Acc65I  (4429)
1 site
G G T A C C C C A T G G
PflFI  (4368)
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  (4368)
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.
BseRI  (3942)
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.
BclI  (3838)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
AscI  (120)
1 site
G G C G C G C C C C G C G C G G
NotI  (148)
2 sites
G C G G C C G C C G C C G G C G
BsrBI  (278)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures up to 50°C.
HindIII  (355)
1 site
A A G C T T T T C G A A
SphI  (365)
1 site
G C A T G C C G T A C G
HpaI  (379)
1 site
G T T A A C C A A T T G
BamHI  (386)
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.
PmlI  (398)
1 site
C A C G T G G T G C A C
EcoRI  (403)
1 site
G A A T T C C T T A A G
KasI  (590)
1 site
G G C G C C C C G C G G
NarI  (591)
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  (592)
1 site
G G C G C C C C G C G G
BbeI  (594)
1 site
G G C G C C C C G C G G

Cleavage may be enhanced when more than one copy of the BbeI recognition sequence is present.
NotI  (620)
2 sites
G C G G C C G C C G C C G G C G
PmeI  (669)
1 site
G T T T A A A C C A A A T T T G
EcoO109I  (983)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (983)
1 site
R G G