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pE-SUMOpro3 Amp

Bacterial vector with an ampicillin resistance marker, for regulated expression of recombinant proteins with a cleavable 6xHis-SUMO3 tag.

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pE-SUMOpro3 Amp.dna
Map and Sequence File:    Download    Open   
Sequence Author:  LifeSensors
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SgrAI (5703) SphI (5555) PflMI (5447) BstAPI (5346) MluI (5022) BclI * (5008) BstEII (4840) ApaI (4819) PspOMI (4815) BssHII (4611) EcoRV (4576) HpaI (4520) PshAI (4181) FspAI (3944) Bpu10I (3816) PflFI - Tth111I (3179) BstZ17I (3154) BspQI - SapI (3038) PciI (2921) BglII (1) XbaI (67) NcoI (106) KflI (133) SbfI (142) BseRI (159) StuI (270) BmgBI (389) BsaI (409) SpeI (421) Acc65I (427) AgeI (430) KpnI (431) BsaI (439) BamHI (445) EcoRI (451) Eco53kI (459) SacI (461) SalI (464) HindIII (470) EagI - NotI (477) AvaI - BsoBI - PaeR7I - PspXI - XhoI (485) BmeT110I (486) DraIII (892) PsiI (1017) ScaI (1552) PvuI (1664) AhdI (2033) AlwNI (2512) pE-SUMOpro3 Amp 5743 bp
SgrAI  (5703)
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.
SphI  (5555)
1 site
G C A T G C C G T A C G
PflMI  (5447)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BstAPI  (5346)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
MluI  (5022)
1 site
A C G C G T T G C G C A
BclI  (5008)
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.
BstEII  (4840)
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.
ApaI  (4819)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4815)
1 site
G G G C C C C C C G G G
BssHII  (4611)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
EcoRV  (4576)
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.
HpaI  (4520)
1 site
G T T A A C C A A T T G
PshAI  (4181)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
FspAI  (3944)
1 site
R T G C G C A Y Y A C G C G T R
Bpu10I  (3816)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
PflFI  (3179)
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  (3179)
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.
BstZ17I  (3154)
1 site
G T A T A C C A T A T G
BspQI  (3038)
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  (3038)
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.
PciI  (2921)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BglII  (1)
1 site
A G A T C T T C T A G A
XbaI  (67)
1 site
T C T A G A A G A T C T
NcoI  (106)
1 site
C C A T G G G G T A C C
KflI  (133)
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.
SbfI  (142)
1 site
C C T G C A G G G G A C G T C C
BseRI  (159)
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.
StuI  (270)
1 site
A G G C C T T C C G G A
BmgBI  (389)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BsaI  (409)
2 sites
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.
SpeI  (421)
1 site
A C T A G T T G A T C A
Acc65I  (427)
1 site
G G T A C C C C A T G G
AgeI  (430)
1 site
A C C G G T T G G C C A
KpnI  (431)
1 site
G G T A C C C C A T G G
BsaI  (439)
2 sites
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.
BamHI  (445)
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.
EcoRI  (451)
1 site
G A A T T C C T T A A G
Eco53kI  (459)
1 site
G A G C T C C T C G A G
SacI  (461)
1 site
G A G C T C C T C G A G
SalI  (464)
1 site
G T C G A C C A G C T G
HindIII  (470)
1 site
A A G C T T T T C G A A
EagI  (477)
1 site
C G G C C G G C C G G C
NotI  (477)
1 site
G C G G C C G C C G C C G G C G
AvaI  (485)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (485)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (485)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (485)
1 site
V C T C G A G B B G A G C T C V
XhoI  (485)
1 site
C T C G A G G A G C T C
BmeT110I  (486)
1 site
C Y C G R G G R G C Y C
DraIII  (892)
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.
PsiI  (1017)
1 site
T T A T A A A A T A T T
ScaI  (1552)
1 site
A G T A C T T C A T G A
PvuI  (1664)
1 site
C G A T C G G C T A G C
AhdI  (2033)
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  (2512)
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.
lacI
4295 .. 5377  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
4295 .. 5377  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
1246 .. 2106  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   1246 .. 1314  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1246 .. 2106  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1315 .. 2106  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1246 .. 2106  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2277 .. 2865  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2277 .. 2865  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
659 .. 1114  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
659 .. 1114  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SUMO3
144 .. 413  =  270 bp
90 amino acids  =  10.3 kDa
Product: human small ubiquitin-related modifier 3
SUMO3
144 .. 413  =  270 bp
90 amino acids  =  10.3 kDa
Product: human small ubiquitin-related modifier 3
rop
3295 .. 3486  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
3295 .. 3486  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
1141 .. 1245  =  105 bp
AmpR promoter
1141 .. 1245  =  105 bp
lacI promoter
5378 .. 5455  =  78 bp
lacI promoter
5378 .. 5455  =  78 bp
MCS
421 .. 490  =  70 bp
multiple cloning site
MCS
421 .. 490  =  70 bp
multiple cloning site
T7 terminator
575 .. 622  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
575 .. 622  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
40 .. 64  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
40 .. 64  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
RBS
79 .. 101  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
79 .. 101  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
114 .. 131  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
114 .. 131  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
491 .. 508  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
491 .. 508  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
108 .. 110  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
108 .. 110  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  1246 .. 2106  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  4261 .. 4512  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  5416 .. 5679  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  3518 .. 3886  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  108 .. 416  =  309 bp
ORF:  102 amino acids  =  11.7 kDa
ORF:  5553 .. 49  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  4295 .. 5254  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3295 .. 3519  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  3883 .. 4239  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  1710 .. 1976  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4275 .. 4538  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  5127 .. 5627  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
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