pEZSeq-Amp

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

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspEZSeq-Amp
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
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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
2 segments
   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
2 segments
   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
3 segments
   Segment 1:  -35  
   18 .. 23  =  6 bp
promoter for the E. coli lac operon
lac promoter
18 .. 48  =  31 bp
3 segments
   Segment 2:  
   24 .. 41  =  18 bp
promoter for the E. coli lac operon
lac promoter
18 .. 48  =  31 bp
3 segments
   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).
ORF:  459 .. 1319  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  923 .. 1225  =  303 bp
ORF:  100 amino acids  =  10.5 kDa
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Individual Sequences & Maps

pAviTag C-His Kan
pEZ BAC
pRANGER-BTB-2
pSMART GC HK (linearized)
pAviTag N-His Kan
pEZ BAC (BamHI linearized)
pRANGER-BTB-3
pSMART GC LK (linearized)
pAviTag-C N-His SUMO Kan
pEZ BAC (HpaI linearized)
pRANGER-BTB-5
pSMART HCAmp
pAviTag-N N-His SUMO Kan
pGC Blue (linearized)
pRham C-His Kan
pSMART HCAmp (linearized)
pETite C-His Kan
pInvRecA
pRham N-His Kan
pSMART HCKan
pETite N-His Kan
pJAZZ-OC
pRham N-His SUMO Kan
pSMART HCKan (linearized)
pETite N-His SUMO Kan
pJAZZ-OK
pSMART BAC v2.0
pSMART LCAmp
pEZSeq-Amp
pJW168
pSMART BAC v2.0 (BamHI linearized)
pSMART LCAmp (linearized)
pEZSeq-Amp (linearized)
pME-HA (linearized)
pSMART BAC v2.0 (EcoRI linearized)
pSMART LCKan
pEZSeq-Kan
pNGS FOS (linearized)
pSMART BAC v2.0 (HindIII linearized)
pSMART LCKan (linearized)
pEZSeq-Kan (linearized)
pRANGER-BTB-1
pSMART FOS v2.0 (linearized)

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