pCOLA-3-DEST

Gateway® Nova bacterial destination vector with a ColA origin for expressing proteins tagged at the N-terminus with Strep-Tag II.

Sequence Author: MilliporeSigma (Novagen)

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EcoO109I (5097) BlpI (5070) AvrII (5052) DraIII (5043) 6xHis SalI (4891) PstI (4889) BfuAI - BspMI (4878) BsaBI * (4647) SrfI (4634) AlwNI (4542) BbvCI (4488) BstZ17I (4298) ScaI (4180) PasI (3995) EcoRI (3763) BspEI (3759) EagI - NotI (3443) BstBI (3300) Strep-Tag II ATG BtgI - NcoI (3273) RBS T7 promoter PfoI (3141) BstAPI (3033) BclI * (2695) BstEII (2527) Bsu36I (2) AgeI (51) DrdI - PflFI - Tth111I (122) BsaI (130) XmnI (218) BspDI - ClaI (914) NruI (950) NheI - NspI - SphI (1139) BmtI (1143) EcoRV (1565) CsiI - SexAI * (1648) PvuI (1740) XbaI (1789) AcuI (1849) KasI (2070) NarI * (2071) SfoI (2072) PluTI (2074) HpaI (2207) PspOMI (2502) ApaI (2506) NmeAIII (2509) pCOLA-3-DEST™ 5134 bp
EcoO109I  (5097)
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.
BlpI  (5070)
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.
AvrII  (5052)
1 site
C C T A G G G G A T C C
DraIII  (5043)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
SalI  (4891)
1 site
G T C G A C C A G C T G
PstI  (4889)
1 site
C T G C A G G A C G T C
BfuAI  (4878)
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  (4878)
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.
BsaBI  (4647)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
SrfI  (4634)
1 site
G C C C G G G C C G G G C C C G
AlwNI  (4542)
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.
BbvCI  (4488)
1 site
C C T C A G C G G A G T C G
BstZ17I  (4298)
1 site
G T A T A C C A T A T G
ScaI  (4180)
1 site
A G T A C T T C A T G A
PasI  (3995)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoRI  (3763)
1 site
G A A T T C C T T A A G
BspEI  (3759)
1 site
T C C G G A A G G C C T
EagI  (3443)
1 site
C G G C C G G C C G G C
NotI  (3443)
1 site
G C G G C C G C C G C