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Plasmid Files

pFLD

Pichia pastoris vector for methylamine-inducible intracellular expression of a protein.

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pFLD.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BglII (4404) MluI (4310) BsrGI (4303) PpuMI (4229) EcoRV (4136) StuI (4102) DraIII (4051) FseI (3974) SexAI * (3861) SgrAI (3812) SmaI (3788) TspMI - XmaI (3786) MauBI (3734) MscI (3701) NcoI - StyI (3696) HaeII (2980) AlwNI (2811) AhdI (2332) BmrI (2292) SphI (5) BbvCI (49) NsiI (284) NdeI (405) BstBI (439) BspDI - ClaI (492) BsaAI - PmlI (608) SfiI (618) Acc65I (634) KpnI (638) PaeR7I - PspXI - XhoI (639) SacII (648) PspOMI (664) ApaI (668) SalI (724) AccI (725) AgeI (806) BsiWI (938) BclI * (954) PvuII (972) BamHI (1087) AscI (1094) SspI (1527) XmnI (1732) PvuI (1963) FspI (2109) pFLD 4409 bp
BglII  (4404)
1 site
A G A T C T T C T A G A
MluI  (4310)
1 site
A C G C G T T G C G C A
BsrGI  (4303)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
PpuMI  (4229)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
EcoRV  (4136)
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.
StuI  (4102)
1 site
A G G C C T T C C G G A
DraIII  (4051)
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.
FseI  (3974)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
SexAI  (3861)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SgrAI  (3812)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SmaI  (3788)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3786)
1 site
C C C G G G G G G C C C
XmaI  (3786)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
MauBI  (3734)
1 site
C G C G C G C G G C G C G C G C
MscI  (3701)
1 site
T G G C C A A C C G G T
NcoI  (3696)
1 site
C C A T G G G G T A C C
StyI  (3696)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
HaeII  (2980)
1 site
R G C G C Y Y C G C G R
AlwNI  (2811)
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.
AhdI  (2332)
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.
BmrI  (2292)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
SphI  (5)
1 site
G C A T G C C G T A C G
BbvCI  (49)
1 site
C C T C A G C G G A G T C G
NsiI  (284)
1 site
A T G C A T T A C G T A
NdeI  (405)
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.
BstBI  (439)
1 site
T T C G A A A A G C T T
BspDI  (492)
1 site
A T C G A T T A G C T A
ClaI  (492)
1 site
A T C G A T T A G C T A
BsaAI  (608)
1 site
Y A C G T R R T G C A Y
PmlI  (608)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
SfiI  (618)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
Acc65I  (634)
1 site
G G T A C C C C A T G G
KpnI  (638)
1 site
G G T A C C C C A T G G
PaeR7I  (639)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (639)
1 site
V C T C G A G B B G A G C T C V
XhoI  (639)
1 site
C T C G A G G A G C T C
SacII  (648)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI  (664)
1 site
G G G C C C C C C G G G
ApaI  (668)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SalI  (724)
1 site
G T C G A C C A G C T G
AccI  (725)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
AgeI  (806)
1 site
A C C G G T T G G C C A
BsiWI  (938)
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.
BclI  (954)
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.
PvuII  (972)
1 site