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Plasmid Files

pZsYellow

Vector for expressing ZsYellow1 in bacteria.

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pZsYellow.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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BspQI - SapI (3323) AflIII - PciI (3206) PspFI (2906) BseYI (2902) AlwNI (2797) BsaI (2252) BpmI (2249) NmeAIII (2171) FspI (2095) PvuI (1949) TsoI (1920) ScaI (1837) XmnI (1718) HindIII (234) PstI - SbfI (250) SalI (252) BamHI (264) AvaI - BsoBI - TspMI - XmaI (269) BmeT110I (270) SmaI (271) Acc65I (273) KpnI (277) BtgI - NcoI (287) BclI * (371) EcoRV (394) Bpu10I (755) EcoNI (799) BseRI (889) BtgZI (932) BsmI (961) NotI (987) EcoRI (1001) StuI (1061) SpeI (1065) BsiWI (1075) PspOMI (1080) ApaI - BanII (1084) PfoI (1138) BstAPI (1277) AflII (1333) ZraI (1397) AatII (1399) SspI (1513) pZsYellow 3331 bp
BspQI  (3323)
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  (3323)
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  (3206)
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  (3206)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2906)
1 site
C C C A G C G G G T C G
BseYI  (2902)
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  (2797)
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  (2252)
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  (2249)
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  (2171)
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  (2095)
1 site
T G C G C A A C G C G T
PvuI  (1949)
1 site
C G A T C G G C T A G C
TsoI  (1920)
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).
ScaI  (1837)
1 site
A G T A C T T C A T G A
XmnI  (1718)
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
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
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
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
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.
Bpu10I  (755)
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.
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.
BseRI  (889)
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.
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.
BsmI  (961)
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.
NotI  (987)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (1001)
1 site
G A A T T C C T T A A G
StuI  (1061)
1 site
A G G C C T T C C G G A
SpeI  (1065)
1 site
A C T A G T T G A T C A
BsiWI  (1075)
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  (1080)
1 site
G G G C C C C C C G G G
ApaI  (1084)
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  (1084)
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  (1138)
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  (1277)
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  (1333)
1 site
C T T A A G G A A T T C
ZraI  (1397)
1 site
G A C G T C C T G C A G
AatII  (1399)
1 site
G A C G T C C T G C A G
SspI  (1513)
1 site
A A T A T T T T A T A A
AmpR
1531 .. 2391  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1531 .. 1599  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1531 .. 2391  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1600 .. 2391  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1531 .. 2391  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ZsYellow
289 .. 984  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus yellow fluorescent protein
ZsYellow
289 .. 984  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus yellow fluorescent protein
ori
2562 .. 3150  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2562 .. 3150  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
1426 .. 1530  =  105 bp
AmpR promoter
1426 .. 1530  =  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).
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
ORF:  217 .. 984  =  768 bp
ORF:  255 amino acids  =  28.7 kDa
ORF:  1531 .. 2391  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  472 .. 954  =  483 bp
ORF:  160 amino acids  =  18.6 kDa
ORF:  1995 .. 2261  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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