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Plasmid Files

pZsGreen

Vector for expressing ZsGreen in bacteria.

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pZsGreen Sequence and MappZsGreen.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BspQI - SapI (3322) AflIII - PciI (3205) PspFI (2905) BseYI (2901) AlwNI (2796) BsaI (2251) BpmI (2248) NmeAIII (2170) FspI (2094) PvuI (1948) ScaI (1836) XmnI (1717) HindIII (234) BfuAI - BspMI (239) SphI (244) PstI - SbfI (250) SalI (252) AccI (253) HincII (254) BamHI (264) AvaI - BsoBI - TspMI - XmaI (269) BmeT110I (270) SmaI (271) Acc65I (273) AgeI (276) KpnI (277) BtgI - NcoI - StyI (287) BclI * (371) EcoRV (394) EcoNI (799) BtgZI (932) BspEI * (964) NotI (986) EcoRI (1000) StuI (1060) SpeI (1064) BsiWI (1074) PspOMI (1079) ApaI - BanII (1083) PfoI (1137) BstAPI (1276) AflII (1332) ZraI (1396) AatII (1398) SspI (1512) pZsGreen 3330 bp
BspQI  (3322)
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  (3322)
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.
AflIII  (3205)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3205)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2905)
1 site
C C C A G C G G G T C G
BseYI  (2901)
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.
AlwNI  (2796)
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.
BsaI  (2251)
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  (2248)
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.
NmeAIII  (2170)
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).
FspI  (2094)
1 site
T G C G C A A C G C G T
PvuI  (1948)
1 site
C G A T C G G C T A G C
ScaI  (1836)
1 site
A G T A C T T C A T G A
XmnI  (1717)
1 site
G A A N N N N T T C C T T N N N N A A G
HindIII  (234)
1 site
A A G C T T T T C G A A
BfuAI  (239)
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  (239)
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.
SphI  (244)
1 site
G C A T G C C G T A C G
PstI  (250)
1 site
C T G C A G G A C G T C
SbfI  (250)
1 site
C C T G C A G G G G A C G T C C
SalI  (252)
1 site
G T C G A C C A G C T G
AccI  (253)
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  (254)
1 site
G T Y R A C C A R Y T G
BamHI  (264)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
AvaI  (269)
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  (269)
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.
TspMI  (269)
1 site
C C C G G G G G G C C C
XmaI  (269)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmeT110I  (270)
1 site
C Y C G R G G R G C Y C
SmaI  (271)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (273)
1 site
G G T A C C C C A T G G
AgeI  (276)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
KpnI  (277)
1 site
G G T A C C C C A T G G
BtgI  (287)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (287)
1 site
C C A T G G G G T A C C
StyI  (287)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BclI  (371)
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.
EcoRV  (394)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
EcoNI  (799)
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.
BtgZI  (932)
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.
BspEI  (964)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
NotI  (986)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (1000)
1 site
G A A T T C C T T A A G
StuI  (1060)
1 site
A G G C C T T C C G G A
SpeI  (1064)
1 site
A C T A G T T G A T C A
BsiWI  (1074)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PspOMI  (1079)
1 site
G G G C C C C C C G G G
ApaI  (1083)
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  (1083)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PfoI  (1137)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstAPI  (1276)
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.
AflII  (1332)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
ZraI  (1396)
1 site
G A C G T C C T G C A G
AatII  (1398)
1 site
G A C G T C C T G C A G
SspI  (1512)
1 site
A A T A T T T T A T A A
AmpR
1530 .. 2390  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1530 .. 1598  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1530 .. 2390  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1599 .. 2390  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1530 .. 2390  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ZsGreen
289 .. 984  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
ZsGreen
289 .. 984  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
ori
2561 .. 3149  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2561 .. 3149  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
1425 .. 1529  =  105 bp
AmpR promoter
1425 .. 1529  =  105 bp
lac promoter
143 .. 173  =  31 bp
   Segment 1:  -35  
   143 .. 148  =  6 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 2:  
   149 .. 166  =  18 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 3:  -10  
   167 .. 173  =  7 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
181 .. 197  =  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
181 .. 197  =  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
205 .. 221  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
205 .. 221  =  17 bp
common sequencing primer, one of multiple similar
variants
ATG
217 .. 219  =  3 bp
1 amino acid  =  149.2 Da
Product: lacZ start codon
ATG
217 .. 219  =  3 bp
1 amino acid  =  149.2 Da
Product: lacZ start codon
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