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Plasmid Files

pEASY®-Blunt3 (linearized)

Cloning vector for blunt-ended PCR products. The cloned PCR product can be excised with EcoRI or NotI.

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pEASY-Blunt3 (linearized).dna
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Sequence Author:  TransBionovo (TransGen Biotech)
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PspOMI (2978) NaeI (2660) NgoMIV (2658) BtgZI (2558) BsaAI - DraIII (2557) PsiI (2429) XmnI (1962) ScaI (1843) TatI (1841) NmeAIII (1511) BpmI (1433) BsaI (1424) ApaI (2982) ZraI (2986) AatII (2988) SphI (2994) NcoI - StyI (3005) NotI (3011) SacII (3017) EcoRI (3020) End (3040) Start (0) BsaBI * (11) SpeI (17) EcoRI (23) BfuAI - BspMI - NotI (30) PstI - SbfI (41) SalI (43) AccI (44) HincII (45) NdeI (50) Eco53kI (60) SacI (62) MluI (67) BstXI (71) NsiI (80) BspQI - SapI (354) PciI (470) BseYI (774) PspFI (778) AlwNI (886) AhdI (1363) pEASY®-Blunt3 3040 bp
PspOMI  (2978)
1 site
G G G C C C C C C G G G
NaeI  (2660)
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.
NgoMIV  (2658)
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.
BtgZI  (2558)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (2557)
1 site
Y A C G T R R T G C A Y
DraIII  (2557)
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.
PsiI  (2429)
1 site
T T A T A A A A T A T T
XmnI  (1962)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1843)
1 site
A G T A C T T C A T G A
TatI  (1841)
1 site
W G T A C W W C A T G W
NmeAIII  (1511)
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).
BpmI  (1433)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsaI  (1424)
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.
ApaI  (2982)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
ZraI  (2986)
1 site
G A C G T C C T G C A G
AatII  (2988)
1 site
G A C G T C C T G C A G
SphI  (2994)
1 site
G C A T G C C G T A C G
NcoI  (3005)
1 site
C C A T G G G G T A C C
StyI  (3005)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NotI  (3011)
2 sites
G C G G C C G C C G C C G G C G
SacII  (3017)
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.
EcoRI  (3020)
2 sites
G A A T T C C T T A A G
End  (3040)
0 sites
Start  (0)
0 sites
BsaBI  (11)
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.
SpeI  (17)
1 site
A C T A G T T G A T C A
EcoRI  (23)
2 sites
G A A T T C C T T A A G
BfuAI  (30)
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  (30)
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.
NotI  (30)
2 sites
G C G G C C G C C G C C G G C G
PstI  (41)
1 site
C T G C A G G A C G T C
SbfI  (41)
1 site
C C T G C A G G G G A C G T C C
SalI  (43)
1 site
G T C G A C C A G C T G
AccI  (44)
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.
HincII  (45)
1 site
G T Y R A C C A R Y T G
NdeI  (50)
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.
Eco53kI  (60)
1 site
G A G C T C C T C G A G
SacI  (62)
1 site
G A G C T C C T C G A G
MluI  (67)
1 site
A C G C G T T G C G C A
BstXI  (71)
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.
NsiI  (80)
1 site
A T G C A T T A C G T A
BspQI  (354)
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  (354)
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.
PciI  (470)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (774)
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.
PspFI  (778)
1 site
C C C A G C G G G T C G
AlwNI  (886)
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.
AhdI  (1363)
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.
AmpR
1290 .. 2150  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1290 .. 2081  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1290 .. 2150  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2082 .. 2150  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1290 .. 2150  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
531 .. 1119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
531 .. 1119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2333 .. 2788  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2333 .. 2788  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
2151 .. 2255  =  105 bp
AmpR promoter
2151 .. 2255  =  105 bp
lac promoter
177 .. 207  =  31 bp
   Segment 3:  -10  
   177 .. 183  =  7 bp
promoter for the E. coli lac operon
lac promoter
177 .. 207  =  31 bp
   Segment 2:  
   184 .. 201  =  18 bp
promoter for the E. coli lac operon
lac promoter
177 .. 207  =  31 bp
   Segment 1:  -35  
   202 .. 207  =  6 bp
promoter for the E. coli lac operon
lac promoter
177 .. 207  =  31 bp
3 segments
promoter for the E. coli lac operon
SP6 promoter
93 .. 111  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
93 .. 111  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 rev
129 .. 145  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
129 .. 145  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
153 .. 169  =  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
153 .. 169  =  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).
M13 fwd
2929 .. 2945  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2929 .. 2945  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1420 .. 1686  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2765 .. 3040  =  276 bp
ORF:  91 amino acids  =  10.4 kDa  (no start codon)
ORF:  2047 .. 2328  =  282 bp
ORF:  93 amino acids  =  10.9 kDa
ORF:  2800 .. 3039  =  240 bp
ORF:  79 amino acids  =  8.4 kDa  (no start codon)
ORF:  1290 .. 2150  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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