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Plasmid Files

pRS403

Yeast integrative vector with a HIS3 marker and an MCS derived from pBLUESCRIPT II.

 
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pRS403.dna
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AatII (4391) ZraI (4389) XmnI (4068) ScaI (3949) NmeAIII (3617) BsaI (3530) AhdI (3469) AlwNI (2992) PspFI (2884) BseYI (2880) BspQI - SapI (2460) PfoI (46) AfeI (213) BseRI (303) BsaBI * (561) NdeI (694) MscI (711) BsmI (716) BbsI (790) BsiWI (916) NheI (1008) BmtI (1012) BclI * (1052) NsiI (1283) PsiI (1519) DraIII (1647) BtgZI (1648) NgoMIV (1748) NaeI (1750) PspOMI (2082) ApaI (2086) AbsI - PaeR7I - PspXI - XhoI (2091) SalI (2097) HincII (2099) BspDI - ClaI (2107) EcoRV (2120) EcoRI (2124) TspMI - XmaI (2136) SmaI (2138) BamHI (2142) SpeI (2148) XbaI (2154) EagI - NotI (2161) BtgI (2170) SacII (2173) Eco53kI (2180) SacI (2182) pRS403 4456 bp
AatII  (4391)
1 site
G A C G T C C T G C A G
ZraI  (4389)
1 site
G A C G T C C T G C A G
XmnI  (4068)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3949)
1 site
A G T A C T T C A T G A
NmeAIII  (3617)
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).
BsaI  (3530)
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  (3469)
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  (2992)
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.
PspFI  (2884)
1 site
C C C A G C G G G T C G
BseYI  (2880)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BspQI  (2460)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2460)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
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.
AfeI  (213)
1 site
A G C G C T T C G C G A
BseRI  (303)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BsaBI  (561)
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.
NdeI  (694)
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.
MscI  (711)
1 site
T G G C C A A C C G G T
BsmI  (716)
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.
BbsI  (790)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsiWI  (916)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
NheI  (1008)
1 site
G C T A G C C G A T C G
BmtI  (1012)
1 site
G C T A G C C G A T C G
BclI  (1052)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
NsiI  (1283)
1 site
A T G C A T T A C G T A
PsiI  (1519)
1 site
T T A T A A A A T A T T
DraIII  (1647)
1 site
C A C N N N G T G G T G