Resources
Plasmid Files

pEASY®-Blunt E1 (linearized)

Prokaryotic expression vector for cloning blunt-ended PCR products and adding an N-terminal 6xHis tag.

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

pEASY-Blunt E1 (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  TransBionovo (TransGen Biotech)
Download Free Trial Get SnapGene Viewer


EcoNI (5301) MluI (4833) BclI * (4819) BstEII (4651) ApaI (4630) PspOMI (4626) ApoI (4558) BssHII (4422) HpaI (4331) PshAI (3992) EagI (3765) NruI (3734) BfuAI - BspMI (3650) BsmI (3351) BmeT110I (3280) AvaI - BsoBI (3279) MscI * (3262) FspAI (3252) Bpu10I (3124) PflFI - Tth111I (2487) BsaAI (2481) BstZ17I (2462) AccI (2461) SphI (5366) SgrAI (5514) BglII (5555) XbaI (5621) NdeI (5661) NheI (5699) BmtI (5703) End (5716) Start (0) Eco53kI (15) SacI (17) BlpI (74) BseRI (135) HindIII (311) BspDI - ClaI (318) ZraI (423) AatII (425) SspI (539) ScaI (863) PvuI (975) PstI (1102) BsaI (1278) AhdI (1344) AlwNI (1823) PciI (2232) BspQI - SapI (2349) pEASY®-Blunt E1 5716 bp
EcoNI  (5301)
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.
MluI  (4833)
1 site
A C G C G T T G C G C A
BclI  (4819)
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.
BstEII  (4651)
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.
ApaI  (4630)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4626)
1 site
G G G C C C C C C G G G
ApoI  (4558)
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.
BssHII  (4422)
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.
HpaI  (4331)
1 site
G T T A A C C A A T T G
PshAI  (3992)
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  (3765)
1 site
C G G C C G G C C G G C
NruI  (3734)
1 site
T C G C G A A G C G C T
BfuAI  (3650)
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  (3650)
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  (3351)
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.
BmeT110I  (3280)
1 site
C Y C G R G G R G C Y C
AvaI  (3279)
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  (3279)
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.
MscI  (3262)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
FspAI  (3252)
1 site
R T G C G C A Y Y A C G C G T R
Bpu10I  (3124)
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.
PflFI  (2487)
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  (2487)
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.
BsaAI  (2481)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2462)
1 site
G T A T A C C A T A T G
AccI  (2461)
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.
SphI  (5366)
1 site
G C A T G C C G T A C G
SgrAI  (5514)
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.
BglII  (5555)
1 site
A G A T C T T C T A G A
XbaI  (5621)
1 site
T C T A G A A G A T C T
NdeI  (5661)
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.
NheI  (5699)
1 site
G C T A G C C G A T C G
BmtI  (5703)
1 site
G C T A G C C G A T C G
End  (5716)
0 sites
Start  (0)
0 sites
Eco53kI  (15)
1 site
G A G C T C C T C G A G
SacI  (17)
1 site
G A G C T C C T C G A G
BlpI  (74)
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.
BseRI  (135)
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.
HindIII  (311)
1 site
A A G C T T T T C G A A
BspDI  (318)
1 site
A T C G A T T A G C T A
ClaI  (318)
1 site
A T C G A T T A G C T A
ZraI  (423)
1 site
G A C G T C C T G C A G
AatII  (425)
1 site
G A C G T C C T G C A G
SspI  (539)
1 site
A A T A T T T T A T A A
ScaI  (863)
1 site
A G T A C T T C A T G A
PvuI  (975)
1 site
C G A T C G G C T A G C
PstI  (1102)
1 site
C T G C A G G A C G T C
BsaI  (1278)
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  (1344)
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  (1823)
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.
PciI  (2232)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2349)
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  (2349)
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.
lacI
4106 .. 5188  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
4106 .. 5188  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
AmpR
557 .. 1417  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   557 .. 625  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
557 .. 1417  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   626 .. 1417  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
557 .. 1417  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1588 .. 2176  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1588 .. 2176  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
2603 .. 2794  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
2603 .. 2794  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
452 .. 556  =  105 bp
AmpR promoter
452 .. 556  =  105 bp
lacI promoter
5189 .. 5266  =  78 bp
lacI promoter
5189 .. 5266  =  78 bp
T7 terminator
85 .. 132  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
85 .. 132  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tet promoter
307 .. 335  =  29 bp
   Segment 3:  -10  
   307 .. 312  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
307 .. 335  =  29 bp
   Segment 2:  
   313 .. 329  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
307 .. 335  =  29 bp
   Segment 1:  -35  
   330 .. 335  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
307 .. 335  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
lac operator
5594 .. 5618  =  25 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
5594 .. 5618  =  25 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).
RBS
5633 .. 5655  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
5633 .. 5655  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
5575 .. 5593  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5575 .. 5593  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
5675 .. 5692  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5675 .. 5692  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
5663 .. 5665  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5663 .. 5665  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  3628 .. 4323  =  696 bp
ORF:  231 amino acids  =  25.4 kDa
ORF:  5227 .. 5490  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  557 .. 1417  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2826 .. 3194  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5364 .. 5603  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  2603 .. 2827  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  3191 .. 3529  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  4106 .. 5065  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  5435 .. 5716  =  282 bp
ORF:  93 amino acids  =  10.0 kDa  (no start codon)
ORF:  1021 .. 1287  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3433 .. 4050  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
ORF:  4086 .. 4349  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4938 .. 5438  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps