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Plasmid Files

pEZSeq™-Amp

Blue/white screening vector with an ampicillin resistance marker for high efficiency blunt end cloning.

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pEZSeq-Amp Sequence and MappEZSeq-Amp.dna
Map and Sequence File   
Sequence Author:  Lucigen
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 Z-Rev (61 .. 84) AlwNI (1737) PspFI (1629) BseYI (1625) BciVI (1524) BspDI - ClaI (1421) ApaI - BanII (1327) EcoO109I - PspOMI (1323) AhdI (1246) BsaI (1180) BsrFI (1161) NmeAIII (1099) AseI (1071) BsrBI (62) lac operator HindIII (109) SalI (115) AccI (116) HincII (117) EcoRI (121) Z-For (147 .. 170) PvuII (213) EarI (222) SwaI (296) BsaAI (365) Bpu10I (445) XmnI (646) BsaHI (706) TatI (763) ScaI (765) TsoI (848) XcmI (863) pEZSeq™-Amp 2056 bp
AlwNI  (1737)
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.
PspFI  (1629)
1 site
C C C A G C G G G T C G
BseYI  (1625)
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.
BciVI  (1524)
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.
BspDI  (1421)
1 site
A T C G A T T A G C T A
ClaI  (1421)
1 site
A T C G A T T A G C T A
ApaI  (1327)
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  (1327)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
EcoO109I  (1323)
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.
PspOMI  (1323)
1 site
G G G C C C C C C G G G
AhdI  (1246)
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  (1180)
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.
BsrFI  (1161)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
NmeAIII  (1099)
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).
AseI  (1071)
1 site
A T T A A T T A A T T A
BsrBI  (62)
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  (109)
1 site
A A G C T T T T C G A A
SalI  (115)
1 site
G T C G A C C A G C T G
AccI  (116)
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  (117)
1 site
G T Y R A C C A R Y T G
EcoRI  (121)
1 site
G A A T T C C T T A A G
PvuII  (213)
1 site
C A G C T G G T C G A C
EarI  (222)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
SwaI  (296)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BsaAI  (365)
1 site
Y A C G T R R T G C A Y
Bpu10I  (445)
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.
XmnI  (646)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (706)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
TatI  (763)
1 site
W G T A C W W C A T G W
ScaI  (765)
1 site
A G T A C T T C A T G A
TsoI  (848)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
XcmI  (863)
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.
Z-Rev
24-mer  /  42% GC
1 binding site
61 .. 84  =  24 annealed bases
Tm  =  58°C
Z-For
24-mer  /  63% GC
1 binding site
147 .. 170  =  24 annealed bases
Tm  =  65°C
AmpR
459 .. 1319  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   459 .. 527  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
459 .. 1319  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   528 .. 1319  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
459 .. 1319  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1382 .. 1969  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1382 .. 1969  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lacZα
92 .. 289  =  198 bp
65 amino acids  =  7.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
92 .. 289  =  198 bp
65 amino acids  =  7.5 kDa
Product: LacZα fragment of β-galactosidase
cat promoter
356 .. 458  =  103 bp
promoter of the E. coli cat gene
cat promoter
356 .. 458  =  103 bp
promoter of the E. coli cat gene
tonB terminator
324 .. 355  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
324 .. 355  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
lac promoter
18 .. 48  =  31 bp
   Segment 1:  -35  
   18 .. 23  =  6 bp
promoter for the E. coli lac operon
lac promoter
18 .. 48  =  31 bp
   Segment 2:  
   24 .. 41  =  18 bp
promoter for the E. coli lac operon
lac promoter
18 .. 48  =  31 bp
   Segment 3:  -10  
   42 .. 48  =  7 bp
promoter for the E. coli lac operon
lac promoter
18 .. 48  =  31 bp
3 segments
promoter for the E. coli lac operon
T3Te terminator
1991 .. 2020  =  30 bp
phage T3 early transcription terminator
T3Te terminator
1991 .. 2020  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1343 .. 1370  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1343 .. 1370  =  28 bp
phage T7 early transcription terminator
lac operator
56 .. 72  =  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
56 .. 72  =  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).
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