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pYES2/NT B

High-copy episomal vector for galactose-inducible expression of N-terminally tagged proteins in S. cerevisiae. For other reading frames, use pYES2/NT A or pYES2/NT C.

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pYES2 NT B.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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NaeI (5894) NgoMIV (5892) SwaI (5566) SnaBI (5262) HpaI (4617) BmgBI (4557) PfoI * (4515) NruI (4211) BlpI (4175) BspDI - ClaI (4031) NdeI (3882) BsgI (3853) SbfI (3819) BfuAI - BspMI (3808) XcmI (3670) BsmI (3437) BsmBI (3171) SpeI (6034) PvuII (471) HindIII (501) Acc65I (602) KpnI (606) Bsu36I (607) BamHI (611) EcoRI (626) EagI - NotI (653) PaeR7I - PspXI - XhoI (659) XbaI (665) PmeI (758) BsrGI (950) BciVI (1422) BspHI (1939) AhdI (2112) BglI (2232) BsaHI (2649) pYES2/NT B 6039 bp
NaeI  (5894)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (5892)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SwaI  (5566)
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.
SnaBI  (5262)
1 site
T A C G T A A T G C A T
HpaI  (4617)
1 site
G T T A A C C A A T T G
BmgBI  (4557)
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.
PfoI  (4515)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
NruI  (4211)
1 site
T C G C G A A G C G C T
BlpI  (4175)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BspDI  (4031)
1 site
A T C G A T T A G C T A
ClaI  (4031)
1 site
A T C G A T T A G C T A
NdeI  (3882)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BsgI  (3853)
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).
SbfI  (3819)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (3808)
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  (3808)
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.
XcmI  (3670)
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.
BsmI  (3437)
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.
BsmBI  (3171)
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.
SpeI  (6034)
1 site
A C T A G T T G A T C A
PvuII  (471)
1 site
C A G C T G G T C G A C
HindIII  (501)
1 site
A A G C T T T T C G A A
Acc65I  (602)
1 site
G G T A C C C C A T G G
KpnI  (606)
1 site
G G T A C C