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Plasmid Files

pEZ™ BAC

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

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pEZ BAC Sequence and MappEZ BAC.dna
Map and Sequence File   
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) 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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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

PmlI gradually loses activity when stored at -20°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

Efficient cleavage requires at least two copies of the BbeI
recognition sequence.
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 W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
DraIII  (1024)
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.
ScaI  (1918)
1 site
A G T A C T T C A T G A
BstAPI  (1998)
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.
BstEII  (2031)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
SexAI  (2035)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
PciI  (2154)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (2155)
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.
PspOMI  (2668)
1 site
G G G C C C C C C G G G
ApaI  (2672)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (2672)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
AccI  (2744)
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.
PfoI  (2884)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SpeI  (2914)
1 site
A C T A G T T G A T C A
AleI  (3097)
1 site
C A C N N N N G T G G T G N N N N C A C
BEZ-F1
24-mer  /  50% GC
1 binding site
238 .. 261  =  24 annealed bases
Tm  =  60°C
BEZ-R1
24-mer  /  54% GC
1 binding site
488 .. 511  =  24 annealed bases
Tm  =  62°C
sopA
3735 .. 4910  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F
plasmid
sopA
3735 .. 4910  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F
plasmid
repE
5489 .. 6244  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the
bacterial F plasmid
repE
5489 .. 6244  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the
bacterial F plasmid
CmR
1288 .. 1947  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1288 .. 1947  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
oriV
6630 .. 7244  =  615 bp
origin of replication for the bacterial F plasmid
oriV
6630 .. 7244  =  615 bp
origin of replication for the bacterial F plasmid
sopC
2218 .. 2691  =  474 bp
centromere-like partitioning region of the bacterial F
plasmid
sopC
2218 .. 2691  =  474 bp
centromere-like partitioning region of the bacterial F
plasmid
cos
764 .. 1162  =  399 bp
lambda cos site; allows packaging into phage
lambda particles
cos
764 .. 1162  =  399 bp
lambda cos site; allows packaging into phage
lambda particles
lacZα
308 .. 580  =  273 bp
90 amino acids  =  10.2 kDa
Product: LacZα fragment of β-galactosidase
lacZα
308 .. 580  =  273 bp
90 amino acids  =  10.2 kDa
Product: LacZα fragment of β-galactosidase
incC
5236 .. 5486  =  251 bp
incompatibility region of the bacterial F plasmid
incC
5236 .. 5486  =  251 bp
incompatibility region of the bacterial F plasmid
ori2
6335 .. 6554  =  220 bp
secondary origin of replication for the bacterial F
plasmid; also known as oriS
ori2
6335 .. 6554  =  220 bp
secondary origin of replication for the bacterial F
plasmid; also known as oriS
loxP
79 .. 112  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
79 .. 112  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
632 .. 665  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
632 .. 665  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
tonB terminator
684 .. 715  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
684 .. 715  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
lac promoter
234 .. 264  =  31 bp
   Segment 1:  -35  
   234 .. 239  =  6 bp
promoter for the E. coli lac operon
lac promoter
234 .. 264  =  31 bp
   Segment 2:  
   240 .. 257  =  18 bp
promoter for the E. coli lac operon
lac promoter
234 .. 264  =  31 bp
   Segment 3:  -10  
   258 .. 264  =  7 bp
promoter for the E. coli lac operon
lac promoter
234 .. 264  =  31 bp
3 segments
promoter for the E. coli lac operon
T3Te terminator
28 .. 57  =  30 bp
phage T3 early transcription terminator
T3Te terminator
28 .. 57  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1962 .. 1985  =  24 bp
phage T7 early transcription terminator
T7Te terminator
1962 .. 1985  =  24 bp
phage T7 early transcription terminator
lac operator
272 .. 288  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
272 .. 288  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
sopB
2764 .. 3735  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F
plasmid
sopB
2764 .. 3735  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F
plasmid
T7 promoter
411 .. 429  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
411 .. 429  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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