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Plasmid Files

pEASY®-T1 Simple (linearized)

TA cloning vector suitable for blue-white selection, with kanamycin and ampicillin resistance markers and a T7 promoter but no multiple cloning site.

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pEASY-T1 Simple (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  TransBionovo (TransGen Biotech)
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PciI (3461) BspHI (2741) AhdI (2573) BsaI (2507) BpmI (2504) ScaI (2092) TatI (2090) XmnI (1973) RsrII (1618) End (3830) Start (1) DraIII (435) PsiI (560) BstAPI (654) StyI (911) XcmI (918) BglII (938) BclI * (943) EagI (1008) PstI (1155) MscI (1184) PflFI - Tth111I (1220) BssHII (1499) SphI (1507) BtgI (1534) pEASY®-T1 Simple 3829 bp
PciI  (3461)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspHI  (2741)
1 site
T C A T G A A G T A C T
AhdI  (2573)
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.
BsaI  (2507)
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.
BpmI  (2504)
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.
ScaI  (2092)
1 site
A G T A C T T C A T G A
TatI  (2090)
1 site
W G T A C W W C A T G W
XmnI  (1973)
1 site
G A A N N N N T T C C T T N N N N A A G
RsrII  (1618)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
End  (3830)
0 sites
Start  (1)
0 sites
DraIII  (435)
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  (560)
1 site
T T A T A A A A T A T T
BstAPI  (654)
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.
StyI  (911)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
XcmI  (918)
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.
BglII  (938)
1 site
A G A T C T T C T A G A
BclI  (943)
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.
EagI  (1008)
1 site
C G G C C G G C C G G C
PstI  (1155)
1 site
C T G C A G G A C G T C
MscI  (1184)
1 site
T G G C C A A C C G G T
PflFI  (1220)
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  (1220)
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.
BssHII  (1499)
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.
SphI  (1507)
1 site
G C A T G C C G T A C G
BtgI  (1534)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
AmpR
1786 .. 2646  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1786 .. 1854  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1786 .. 2646  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1855 .. 2646  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1786 .. 2646  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
974 .. 1768  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
974 .. 1768  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2817 .. 3405  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2817 .. 3405  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
202 .. 630  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
202 .. 630  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
lac promoter
3729 .. 3759  =  31 bp
   Segment 1:  -35  
   3729 .. 3734  =  6 bp
promoter for the E. coli lac operon
lac promoter
3729 .. 3759  =  31 bp
   Segment 2:  
   3735 .. 3752  =  18 bp
promoter for the E. coli lac operon
lac promoter
3729 .. 3759  =  31 bp
   Segment 3:  -10  
   3753 .. 3759  =  7 bp
promoter for the E. coli lac operon
lac promoter
3729 .. 3759  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
20 .. 38  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
20 .. 38  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
45 .. 61  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
45 .. 61  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
3767 .. 3783  =  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
3767 .. 3783  =  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 rev
3791 .. 3807  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
3791 .. 3807  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1 .. 225  =  225 bp
ORF:  74 amino acids  =  8.5 kDa  (no start codon)
ORF:  1786 .. 2646  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  974 .. 1768  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1146 .. 1532  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2250 .. 2516  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1036 .. 1395  =  360 bp
ORF:  119 amino acids  =  12.5 kDa
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