Resources
Plasmid Files

pHY300PLK

Escherichia coli-Bacillus subtilis shuttle vector with ampicillin and tetracycline resistance genes.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pHY300PLK Sequence and MappHY300PLK.dna
Map and Sequence File   
Sequence Author:  TaKaRa
Download Free Trial Get SnapGene Viewer

 HindIII (1) AgeI (4709) AlwNI (4523) BstBI (4384) BanI (4123) AhdI (4076) BglI (3958) NmeAIII (3929) FspI (3853) PvuI (3707) BaeGI - Bme1580I * (3408) ApaLI (3404) BsrBI (3234) SacII (3186) AatII (3157) ZraI (3155) AflII (3094) XbaI (6) BglII (11) SalI (21) BamHI (27) AvaI - BsoBI - TspMI - XmaI (32) BmeT110I (33) SmaI (34) EcoRI (37) BstEII (136) AflIII (501) EcoO109I - PpuMI (512) PfoI (724) EcoRV (876) PflMI (1007) BspDI * - ClaI * (1038) BssHII (1087) BsgI (1110) MscI (1180) HpaI (1297) AarI (1322) BsmBI (1364) NsiI (2137) BsmI (2381) pHY300PLK 4870 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
AgeI  (4709)
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.
AlwNI  (4523)
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.
BstBI  (4384)
1 site
T T C G A A A A G C T T
BanI  (4123)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
AhdI  (4076)
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.
BglI  (3958)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
NmeAIII  (3929)
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  (3853)
1 site
T G C G C A A C G C G T
PvuI  (3707)
1 site
C G A T C G G C T A G C
BaeGI  (3408)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (3408)
1 site
G K G C M C C M C G K G
* Blocked by EcoKI methylation.
Sticky ends from different Bme1580I sites may not be compatible.
ApaLI  (3404)
1 site
G T G C A C C A C G T G
BsrBI  (3234)
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.
SacII  (3186)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
AatII  (3157)
1 site
G A C G T C C T G C A G
ZraI  (3155)
1 site
G A C G T C C T G C A G
AflII  (3094)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
XbaI  (6)
1 site
T C T A G A A G A T C T
BglII  (11)
1 site
A G A T C T T C T A G A
SalI  (21)
1 site
G T C G A C C A G C T G
BamHI  (27)
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  (32)
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  (32)
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  (32)
1 site
C C C G G G G G G C C C
XmaI  (32)
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  (33)
1 site
C Y C G R G G R G C Y C
SmaI  (34)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (37)
1 site
G A A T T C C T T A A G
BstEII  (136)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
AflIII  (501)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
EcoO109I  (512)
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.
PpuMI  (512)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
PfoI  (724)
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.
EcoRV  (876)
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.
PflMI  (1007)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BspDI  (1038)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1038)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BssHII  (1087)
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.
BsgI  (1110)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
MscI  (1180)
1 site
T G G C C A A C C G G T
HpaI  (1297)
1 site
G T T A A C C A A T T G
AarI  (1322)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BsmBI  (1364)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
NsiI  (2137)
1 site
A T G C A T T A C G T A
BsmI  (2381)
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.
TcR
100 .. 1476  =  1377 bp
458 amino acids  =  50.1 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
100 .. 1476  =  1377 bp
458 amino acids  =  50.1 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
repB
1683 .. 2687  =  1005 bp
334 amino acids  =  39.8 kDa
Product: RepB replication protein
from Enterococcus faecalis plasmid pAMα1
repB
1683 .. 2687  =  1005 bp
334 amino acids  =  39.8 kDa
Product: RepB replication protein
from Enterococcus faecalis plasmid pAMα1
AmpR
3289 .. 4149  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3289 .. 3357  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3289 .. 4149  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3358 .. 4149  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3289 .. 4149  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
p15A ori
4314 .. 4858  =  545 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
p15A ori
4314 .. 4858  =  545 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
AmpR promoter
3184 .. 3288  =  105 bp
AmpR promoter
3184 .. 3288  =  105 bp
MCS
1 .. 42  =  42 bp
multiple cloning site
MCS
1 .. 42  =  42 bp
multiple cloning site
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter