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Plasmid Files

pESC-TRP

Yeast episomal vector with a TRP1 marker, for galactose-regulated expression and tagging of up to two genes.

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pESC-TRP.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Agilent Technologies
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XcmI (6405) BmgBI (6194) NsiI (5404) ScaI (4676) BpmI (4266) BsaI (4257) AhdI (4196) AlwNI (3719) PciI (3303) PfoI (46) BspEI * (332) PmlI (427) Bsu36I (709) EcoRV (752) BsgI (880) AarI - BfuAI - BspMI (900) DraIII (1465) NgoMIV (1566) NaeI (1568) PacI (2078) Eco53kI (2084) SacI (2086) BglII (2088) BspDI - ClaI (2118) SpeI (2123) EagI - NotI (2130) EcoRI (2152) BstBI (2156) AgeI (2447) BamHI (2830) PspOMI (2854) TspMI - XmaI (2857) ApaI (2858) SmaI - SrfI (2859) SalI (2864) PaeR7I - PspXI - XhoI (2900) Acc65I (2913) BtgI - KpnI (2917) SacII (2920) NheI (2922) BmtI (2926) PpuMI (2977) BsrGI (3051) MluI (3058) BspQI - SapI (3187) pESC-TRP 6525 bp
XcmI  (6405)
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.
BmgBI  (6194)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
NsiI  (5404)
1 site
A T G C A T T A C G T A
ScaI  (4676)
1 site
A G T A C T T C A T G A
BpmI  (4266)
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.
BsaI  (4257)
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.
AhdI  (4196)
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.
AlwNI  (3719)
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.
PciI  (3303)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PfoI  (46)
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.
BspEI  (332)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PmlI  (427)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
Bsu36I  (709)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
EcoRV  (752)
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.
BsgI  (880)
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).
AarI  (900)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4