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Plasmid Files

pI-secSUMOstar

Insect cell vector for expression of secreted recombinant proteins with a cleavable 6xHis-SUMOstar tag.

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pI-secSUMOstar.dna
Map and Sequence File:    Download    Open   
Sequence Author:  LifeSensors
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NgoMIV (127) AvrII (4804) HpaI (4653) MfeI (4640) HindIII (4525) KpnI (4523) Acc65I (4519) NotI (4512) BsmBI - XbaI (4505) BsmBI (4481) BlpI (4220) PstI - SbfI (4196) EcoO109I - KflI - PpuMI (4187) BfuAI - BspMI (4147) AfeI (4104) SpeI (4047) BstZ17I (3884) AccI (3883) SnaBI (3881) BbsI (3757) BsrGI (3410) PflFI - Tth111I (3228) BseRI (2961) EcoRV (2825) BstXI (2717) NaeI (129) BanII (161) BsaHI (836) PvuI (1007) FspI (1153) BpmI (1307) AlwNI (1855) BseYI (1960) PspFI (1964) BspQI - SapI (2381) MscI (2709) pI-secSUMOstar 5164 bp
NgoMIV  (127)
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.
AvrII  (4804)
1 site
C C T A G G G G A T C C
HpaI  (4653)
1 site
G T T A A C C A A T T G
MfeI  (4640)
1 site
C A A T T G G T T A A C
HindIII  (4525)
1 site
A A G C T T T T C G A A
KpnI  (4523)
1 site
G G T A C C C C A T G G
Acc65I  (4519)
1 site
G G T A C C C C A T G G
NotI  (4512)
1 site
G C G G C C G C C G C C G G C G
BsmBI  (4505)
2 sites
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.
XbaI  (4505)
1 site
T C T A G A A G A T C T
BsmBI  (4481)
2 sites
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.
BlpI  (4220)
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.
PstI  (4196)
1 site
C T G C A G G A C G T C
SbfI  (4196)
1 site
C C T G C A G G G G A C G T C C
EcoO109I  (4187)
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.
KflI  (4187)
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  (4187)
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.
BfuAI  (4147)
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  (4147)
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.
AfeI  (4104)
1 site
A G C G C T T C G C G A
SpeI  (4047)
1 site
A C T A G T T G A T C A
BstZ17I  (3884)
1 site
G T A T A C C A T A T G
AccI  (3883)
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.
SnaBI  (3881)
1 site
T A C G T A A T G C A T
BbsI  (3757)
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.
BsrGI  (3410)
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.
PflFI  (3228)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3228)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BseRI  (2961)
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.
EcoRV  (2825)
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.
BstXI  (2717)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
NaeI  (129)
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.
BanII  (161)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BsaHI  (836)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
PvuI  (1007)
1 site
C G A T C G G C T A G C
FspI  (1153)
1 site
T G C G C A A C G C G T
BpmI  (1307)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AlwNI  (1855)
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.
BseYI  (1960)
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.
PspFI  (1964)
1 site
C C C A G C G G G T C G
BspQI  (2381)
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  (2381)
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.
MscI  (2709)
1 site
T G G C C A A C C G G T
AmpR
589 .. 1449  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   589 .. 657  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
589 .. 1449  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   658 .. 1449  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
589 .. 1449  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1620 .. 2208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1620 .. 2208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
2802 .. 3335  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
2802 .. 3335  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
2 .. 457  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2 .. 457  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SUMO
4198 .. 4485  =  288 bp
96 amino acids  =  11.0 kDa
Product: cleavable ubiquitin-like protein tag
SUMO
4198 .. 4485  =  288 bp
96 amino acids  =  11.0 kDa
Product: cleavable ubiquitin-like protein tag
Tn7R
2511 .. 2735  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
2511 .. 2735  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
4817 .. 4982  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
4817 .. 4982  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
4654 .. 4788  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4654 .. 4788  =  135 bp
SV40 polyadenylation signal
AmpR promoter
484 .. 588  =  105 bp
AmpR promoter
484 .. 588  =  105 bp
polyhedrin promoter
3904 .. 3995  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
3904 .. 3995  =  92 bp
promoter for the baculovirus polyhedrin gene