Resources
Plasmid Files

pBR322

First-generation E. coli vector for DNA cloning. This plasmid has a low copy number (~20 copies per cell) due to the rop gene.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pBR322 Sequence and MappBR322.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
Download Free Trial Get SnapGene Viewer

 ApoI - EcoRI (4359) AatII (4288) ZraI (4286) SspI (4170) ScaI (3846) PvuI (3736) PstI (3611) AseI (3538) BsaI (3427) AhdI (3366) AlwNI (2889) AflIII - PciI (2473) BspQI - SapI (2357) NdeI (2296) BstZ17I (2246) BsaAI (2227) PflFI - Tth111I (2220) BspDI - ClaI (24) HindIII (29) EcoRV (187) NheI (229) BmtI (233) BamHI (375) SgrAI (410) SphI (566) EcoNI (626) SalI (651) PshAI (716) EagI (939) NruI (974) BfuAI - BspMI (1054) BsmI (1359) StyI (1369) AvaI - BsoBI (1425) BmeT110I (1426) MscI * (1446) Bpu10I (1581) BsgI * (1635) BspEI * (1664) BsaBI * (1672) PvuII (2066) BsmBI (2116) PfoI (2117) pBR322 4361 bp
ApoI  (4359)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (4359)
1 site
G A A T T C C T T A A G
AatII  (4288)
1 site
G A C G T C C T G C A G
ZraI  (4286)
1 site
G A C G T C C T G C A G
SspI  (4170)
1 site
A A T A T T T T A T A A
ScaI  (3846)
1 site
A G T A C T T C A T G A
PvuI  (3736)
1 site
C G A T C G G C T A G C
PstI  (3611)
1 site
C T G C A G G A C G T C
AseI  (3538)
1 site
A T T A A T T A A T T A
BsaI  (3427)
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  (3366)
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  (2889)
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.
AflIII  (2473)
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  (2473)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2357)
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  (2357)
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.
NdeI  (2296)
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.
BstZ17I  (2246)
1 site
G T A T A C C A T A T G
BsaAI  (2227)
1 site
Y A C G T R R T G C A Y
PflFI  (2220)
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  (2220)
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.
BspDI  (24)
1 site
A T C G A T T A G C T A
ClaI  (24)
1 site
A T C G A T T A G C T A
HindIII  (29)
1 site
A A G C T T T T C G A A
EcoRV  (187)
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.
NheI  (229)
1 site
G C T A G C C G A T C G
BmtI  (233)
1 site
G C T A G C C G A T C G
BamHI  (375)
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.
SgrAI  (410)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
SphI  (566)
1 site
G C A T G C C G T A C G
EcoNI  (626)
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.
SalI  (651)
1 site
G T C G A C C A G C T G
PshAI  (716)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
EagI  (939)
1 site
C G G C C G G C C G G C
NruI  (974)
1 site
T C G C G A A G C G C T
BfuAI  (1054)
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  (1054)
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.
BsmI  (1359)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
StyI  (1369)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
AvaI  (1425)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1425)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
BmeT110I  (1426)
1 site
C Y C G R G G R G C Y C
MscI  (1446)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
Bpu10I  (1581)
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.
BsgI  (1635)
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  (1664)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
BsaBI  (1672)
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.
PvuII  (2066)
1 site
C A G C T G G T C G A C
BsmBI  (2116)
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.
PfoI  (2117)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
TcR
86 .. 1276  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
86 .. 1276  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
AmpR
3293 .. 4153  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3293 .. 4084  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3293 .. 4153  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4085 .. 4153  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3293 .. 4153  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2534 .. 3122  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2534 .. 3122  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rop
1915 .. 2106  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
1915 .. 2106  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
bom
2208 .. 2348  =  141 bp
basis of mobility region from pBR322
bom
2208 .. 2348  =  141 bp
basis of mobility region from pBR322
AmpR promoter
4154 .. 4258  =  105 bp
AmpR promoter
4154 .. 4258  =  105 bp
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter