pE-SUMOpro3 Kan

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

Sequence Author: LifeSensors

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T7 promoter BglII (1) SgrAI (5570) SphI (5422) BstAPI (5213) MluI (4889) BclI * (4875) BstEII (4707) NmeAIII (4689) ApaI (4686) PspOMI (4682) BssHII (4478) EcoRV (4443) HpaI (4387) PshAI (4048) BglI (3832) FspI - FspAI (3811) PflFI - Tth111I (3046) BstZ17I (3021) TatI (2984) BspQI - SapI (2905) XbaI (67) RBS NcoI (106) ATG 6xHis KflI (133) PstI - SbfI (142) BseRI (159) StuI (270) BmgBI (389) BsaI (409) SpeI (421) Acc65I (427) AgeI (430) KpnI (431) BsaI (439) BamHI (445) EcoRI (451) MCS Eco53kI (459) SacI (461) SalI (464) HindIII (470) EagI - NotI (477) PaeR7I - PspXI - XhoI (485) DraIII (892) PsiI (1017) AsiSI - PvuI (1592) TspMI - XmaI (1714) SmaI (1716) BspDI - ClaI (1897) NruI (1933) AlwNI (2379) BssSαI (2615) PciI (2788) pE-SUMOpro3 Kan 5610 bp
BglII  (1)
1 site
A G A T C T T C T A G A
SgrAI  (5570)
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  (5422)
1 site
G C A T G C C G T A C G
BstAPI  (5213)
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  (4889)
1 site
A C G C G T T G C G C A
BclI  (4875)
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  (4707)
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.
NmeAIII  (4689)
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).
ApaI  (4686)
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  (4682)
1 site
G G G C C C C C C G G G
BssHII  (4478)
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  (4443)
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  (4387)
1 site
G T T A A C C A A T T G
PshAI  (4048)
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.
BglI  (3832)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
FspI  (3811)
1 site
T G C G C A A C G C G T
FspAI  (3811)
1 site
R T G C G C A Y Y A C G C G T R
PflFI  (3046)
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  (3046)
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  (3021)
1 site
G T A T A C C A T A T G
TatI  (2984)
1 site
W G T A C W W C A T G W
BspQI  (2905)
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  (2905)
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.
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.
PstI  (142)
1 site
C T G C A G G A C G T C
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
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
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
AsiSI  (1592)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1592)
1 site
C G A T C G G C T A G C
TspMI  (1714)
1 site
C C C G G G G G G C C C
XmaI  (1714)
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  (1716)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BspDI  (1897)
1 site
A T C G A T T A G C T A
ClaI  (1897)
1 site
A T C G A T T A G C T A
NruI  (1933)
1 site
T C G C G A A G C G C T
AlwNI  (2379)
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.
BssSαI  (2615)
1 site
C A C G A G G T G C T C
PciI  (2788)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
lacI
4162 .. 5244  =  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
4162 .. 5244  =  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).
KanR
1207 .. 2022  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1207 .. 2022  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
2144 .. 2732  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2144 .. 2732  =  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
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
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
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
3162 .. 3353  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
3162 .. 3353  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
lacI promoter
5245 .. 5322  =  78 bp
lacI promoter
5245 .. 5322  =  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
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
ORF:  3385 .. 3753  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5420 .. 49  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  108 .. 416  =  309 bp
ORF:  102 amino acids  =  11.7 kDa
ORF:  4128 .. 4379  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  5283 .. 5546  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  1207 .. 2022  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  4162 .. 5121  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3162 .. 3386  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  3750 .. 4106  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  4142 .. 4405  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4994 .. 5494  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
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