Resources
Plasmid Files

pBEN1-SGC

Bacterial vector encoding an N-terminal SET1-SBP-TEV cassette plus a SacB negative selection marker, for purification of recombinant proteins.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pBEN1-SGC Sequence and MappBEN1-SGC.dna
Map and Sequence File   
Sequence Author:  Structural Genomics Consortium
Download Free Trial Get SnapGene Viewer

 XbaI (7481) lac operator BglII (7415) SgrAI (7374) SphI (7226) BstAPI (7017) MluI (6693) BclI * (6679) BstEII (6511) NmeAIII (6493) ApaI (6490) PspOMI (6486) BssHII (6282) FspI - FspAI (5615) PpuMI (5587) PflFI - Tth111I (4850) BspQI - SapI (4709) AlwNI (4183) NruI (3737) NdeI (6) ATG SpeI (137) PmlI (175) Bsu36I (234) BsaI (302) NcoI (303) ZraI (316) AatII (318) BfuAI - BspMI (336) sacB promoter StuI * (1007) MscI (1029) AanI - AanI (1406) BsrGI (1446) SnaBI (1515) SacII (1845) BsaI (2233) BamHI (2249) EcoRI (2255) Eco53kI (2263) SacI (2265) SalI (2268) HindIII (2274) EagI - NotI (2281) PaeR7I - PspXI - XhoI (2289) BlpI (2368) AanI - AanI (2821) AsiSI - PvuI (3396) TspMI - XmaI (3518) SmaI (3520) pBEN1-SGC 7515 bp
XbaI  (7481)
1 site
T C T A G A A G A T C T
BglII  (7415)
1 site
A G A T C T T C T A G A
SgrAI  (7374)
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  (7226)
1 site
G C A T G C C G T A C G
BstAPI  (7017)
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  (6693)
1 site
A C G C G T T G C G C A
BclI  (6679)
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  (6511)
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  (6493)
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  (6490)
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  (6486)
1 site
G G G C C C C C C G G G
BssHII  (6282)
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.
FspI  (5615)
1 site
T G C G C A A C G C G T
FspAI  (5615)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (5587)
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.
PflFI  (4850)
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  (4850)
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.
BspQI  (4709)
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  (4709)
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.
AlwNI  (4183)
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.
NruI  (3737)
1 site
T C G C G A A G C G C T
NdeI  (6)
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.
SpeI  (137)
1 site
A C T A G T T G A T C A
PmlI  (175)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
Bsu36I  (234)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BsaI  (302)
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.
NcoI  (303)
1 site
C C A T G G G G T A C C
ZraI  (316)
1 site
G A C G T C C T G C A G
AatII  (318)
1 site
G A C G T C C T G C A G
BfuAI  (336)
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  (336)
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.
StuI  (1007)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
MscI  (1029)
1 site
T G G C C A A C C G G T
AanI  (1406)
2 sites
T T A T A A A A T A T T
AanI  (1406)
2 sites
T T A T A A A A T A T T
BsrGI  (1446)
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.
SnaBI  (1515)
1 site
T A C G T A A T G C A T
SacII  (1845)
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.
BsaI  (2233)
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  (2249)
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  (2255)
1 site
G A A T T C C T T A A G
Eco53kI  (2263)
1 site
G A G C T C C T C G A G
SacI  (2265)
1 site
G A G C T C C T C G A G
SalI  (2268)
1 site
G T C G A C C A G C T G
HindIII  (2274)
1 site
A A G C T T T T C G A A
EagI  (2281)
1 site
C G G C C G G C C G G C
NotI  (2281)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (2289)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2289)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2289)
1 site
C T C G A G G A G C T C
BlpI  (2368)
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.
AanI  (2821)
2 sites
T T A T A A A A T A T T
AanI  (2821)
2 sites
T T A T A A A A T A T T
AsiSI  (3396)
1 site
G C G A T C G C C G C T A G C G
PvuI  (3396)
1 site
C G A T C G G C T A G C
TspMI  (3518)
1 site
C C C G G G G G G C C C
XmaI  (3518)
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.
SmaI  (3520)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SacB
766 .. 2187  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 1:  signal peptide  
   766 .. 852  =  87 bp
   29 amino acids  =  3.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
766 .. 2187  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 2:  
   853 .. 2187  =  1335 bp
   444 amino acids  =  50.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
766 .. 2187  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
lacI
5966 .. 7048  =  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
5966 .. 7048  =  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
3011 .. 3826  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
3011 .. 3826  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
ori
3948 .. 4536  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3948 .. 4536  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2463 .. 2918  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2463 .. 2918  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
sacB promoter
320 .. 765  =  446 bp
sacB promoter and control region
sacB promoter
320 .. 765  =  446 bp
sacB promoter and control region
rop
4966 .. 5157  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
4966 .. 5157  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
SET1
11 .. 136  =  126 bp
42 amino acids  =  4.4 kDa
Product: solubility-enhancement tag derived from
T7 phage gene 10B
SET1
11 .. 136  =  126 bp
42 amino acids  =  4.4 kDa
Product: solubility-enhancement tag derived from
T7 phage gene 10B
SBP
143 .. 253  =  111 bp
37 amino acids  =  4.2 kDa
Product: streptavidin-binding peptide
selected from a peptide library; binds streptavidin
with nanomolar affinity
SBP
143 .. 253  =  111 bp
37 amino acids  =  4.2 kDa
Product: streptavidin-binding peptide
selected from a peptide library; binds streptavidin
with nanomolar affinity
lacI promoter
7049 .. 7126  =  78 bp
lacI promoter
7049 .. 7126  =  78 bp
T7 terminator
2379 .. 2426  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
2379 .. 2426  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
7454 .. 7478  =  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
7454 .. 7478  =  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).
TEV site
284 .. 304  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
284 .. 304  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
7435 .. 7453  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
7435 .. 7453  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
2295 .. 2312  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2295 .. 2312  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
8 .. 10  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
8 .. 10  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter