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pYES2/NT A

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

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pYES2 NT A.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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NaeI (5893) NgoMIV (5891) SwaI (5565) SnaBI (5261) HpaI (4616) BmgBI (4556) PfoI * (4514) NruI (4210) BlpI (4174) BspDI - ClaI (4030) NdeI (3881) BsgI (3852) SbfI (3818) BfuAI - BspMI (3807) XcmI (3669) BsmI (3436) BsmBI (3170) SpeI (6033) PvuII (471) HindIII (501) Acc65I (602) AvrII - KpnI (606) BamHI (610) EcoRI (625) EagI - NotI (652) PaeR7I - PspXI - XhoI (658) XbaI (664) PmeI (757) BsrGI (949) BciVI (1421) BspHI (1938) AhdI (2111) BglI (2231) BsaHI (2648) pYES2/NT A 6038 bp
NaeI  (5893)
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  (5891)
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  (5565)
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  (5261)
1 site
T A C G T A A T G C A T
HpaI  (4616)
1 site
G T T A A C C A A T T G
BmgBI  (4556)
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  (4514)
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  (4210)
1 site
T C G C G A A G C G C T
BlpI  (4174)
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  (4030)
1 site
A T C G A T T A G C T A
ClaI  (4030)
1 site
A T C G A T T A G C T A
NdeI  (3881)
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  (3852)
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  (3818)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (3807)
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  (3807)
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  (3669)
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  (3436)
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  (3170)
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  (6033)
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