pVL-FH

Baculovirus transfer plasmid for high-level expression of proteins with a C-terminal FLAG®-8xHis cassette.

Sequence Author: AB Vector

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NdeI (9509) ScaI (8818) AlwNI (7861) PspFI (7753) BseYI (7749) BspQI - SapI (7329) FspAI (6412) SgrDI (6259) SgrAI (5990) SnaBI (4902) KpnI (4722) PasI (486) PacI (583) XcmI (746) BtgI (868) SacII (871) BstEII (923) PspOMI (1395) ApaI - BanII (1399) PaeR7I - PspXI - XhoI (1901) SphI (2135) BclI * (2232) BbvCI - Bpu10I (3405) NgoMIV (3770) NaeI (3772) EcoRV (4000) BglII (4134) PstI (4142) EagI - NotI (4144) EcoRI (4155) TspMI - XmaI (4160) SmaI (4162) BamHI (4164) XbaI (4172) AvrII (4246) KflI - PpuMI (4271) Acc65I (4718) pVL-FH 9723 bp
NdeI  (9509)
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.
ScaI  (8818)
1 site
A G T A C T T C A T G A
AlwNI  (7861)
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  (7753)
1 site
C C C A G C G G G T C G
BseYI  (7749)
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  (7329)
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  (7329)
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.
FspAI  (6412)
1 site
R T G C G C A Y Y A C G C G T R
SgrDI  (6259)
1 site
C G T C G A C G G C A G C T G C
SgrAI  (5990)
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.
SnaBI  (4902)
1 site
T A C G T A A T G C A T
KpnI  (4722)
1 site
G G T A C C C C A T G G
PasI  (486)
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.
PacI  (583)
1 site
T T A A T T A A A A T T A A T T
XcmI  (746)
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.
BtgI  (868)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (871)
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.
BstEII  (923)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PspOMI  (1395)
1 site
G G G C C C C C C G G G
ApaI  (1399)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (1399)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PaeR7I  (1901)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1901)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1901)
1 site
C T C G A G G A G C T C
SphI  (2135)
1 site
G C A T G C C G T A C G
BclI  (2232)
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.
BbvCI  (3405)
1 site
C C T C A G C G G A G T C G
Bpu10I  (3405)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
NgoMIV  (3770)
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.
NaeI  (3772)
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.
EcoRV  (4000)
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.
BglII  (4134)
1 site
A G A T C T T C T A G A
PstI  (4142)
1 site
C T G C A G G A C G T C
EagI  (4144)
1 site
C G G C C G G C C G G C
NotI  (4144)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (4155)
1 site
G A A T T C C T T A A G
TspMI  (4160)
1 site
C C C G G G G G G C C C
XmaI  (4160)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (4162)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (4164)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
XbaI  (4172)
1 site
T C T A G A A G A T C T
AvrII  (4246)
1 site
C C T A G G G G A T C C
KflI  (4271)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (4271)
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.
Acc65I  (4718)
1 site