Resources
Plasmid Files

pET-61-DEST™

Gateway® Nova bacterial destination vector for expressing proteins tagged at the N-terminus with 6xHis and Strep-Tag II.

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

pET-61-DEST Sequence and MappET-61-DEST.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
Download Free Trial Get SnapGene Viewer

 AvrII (6745) SalI (6594) PstI (6592) BfuAI - BspMI (6581) BmgBI (6371) SmaI - SrfI (6337) BmeT110I (6336) AvaI - BsoBI - TspMI - XmaI (6335) BbvCI (6191) PasI (5698) EcoRI (5466) EagI - NotI (5146) BstBI (5003) Strep-Tag II ATG NcoI (4955) XbaI (4916) lac operator T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) HpaI (3622) BlpI (6763) DraIII (245) PsiI (370) AhdI (655) BsaI (716) BglI (775) FspI (877) PvuI (1025) PciI (2260) BspQI - SapI (2377) PflFI - Tth111I (2518) PpuMI (3255) pET-61-DEST™ 6846 bp
AvrII  (6745)
1 site
C C T A G G G G A T C C
SalI  (6594)
1 site
G T C G A C C A G C T G
PstI  (6592)
1 site
C T G C A G G A C G T C
BfuAI  (6581)
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  (6581)
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.
BmgBI  (6371)
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.
SmaI  (6337)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (6337)
1 site
G C C C G G G C C G G G C C C G
BmeT110I  (6336)
1 site
C Y C G R G G R G C Y C
AvaI  (6335)
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  (6335)
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.
TspMI  (6335)
1 site
C C C G G G G G G C C C
XmaI  (6335)
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.
BbvCI  (6191)
1 site
C C T C A G C G G A G T C G
PasI  (5698)
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.
EcoRI  (5466)
1 site
G A A T T C C T T A A G
EagI  (5146)
1 site
C G G C C G G C C G G C
NotI  (5146)
1 site
G C G G C C G C C G C C G G C G
BstBI  (5003)
1 site
T T C G A A A A G C T T
NcoI  (4955)
1 site
C C A T G G G G T A C C
XbaI  (4916)
1 site
T C T A G A A G A T C T
BspDI  (4849)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4849)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (4805)
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  (4657)
1 site
G C A T G C C G T A C G
EcoNI  (4592)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BstAPI  (4448)
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.
BclI  (4110)
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  (3942)
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  (3921)
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  (3917)
1 site
G G G C C C C C C G G G
HpaI  (3622)
1 site
G T T A A C C A A T T G
BlpI  (6763)
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.
DraIII  (245)
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  (370)
1 site
T T A T A A A A T A T T
AhdI  (655)
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.
BsaI  (716)
1 site
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.
BglI  (775)
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  (877)
1 site
T G C G C A A C G C G T
PvuI  (1025)
1 site
C G A T C G G C T A G C
PciI  (2260)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2377)
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  (2377)
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.
PflFI  (2518)
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  (2518)
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.
PpuMI  (3255)
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.
lacI
3397 .. 4479  =  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
3397 .. 4479  =  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
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   582 .. 1373  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1374 .. 1442  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
5253 .. 5912  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5253 .. 5912  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
ccdB
6254 .. 6559  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
6254 .. 6559  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
rop
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
attR1
5020 .. 5144  =  125 bp
recombination site for the Gateway® LR reaction
attR1
5020 .. 5144  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6600 .. 6724  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6600 .. 6724  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
1443 .. 1535  =  93 bp
AmpR promoter
1443 .. 1535  =  93 bp
lacI promoter
4480 .. 4557  =  78 bp
lacI promoter
4480 .. 4557  =  78 bp
T7 terminator
6774 .. 6821  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
6774 .. 6821  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac UV5 promoter
5169 .. 5199  =  31 bp
   Segment 1:  -35  
   5169 .. 5174  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5169 .. 5199  =  31 bp
   Segment 2:  
   5175 .. 5192  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5169 .. 5199  =  31 bp
   Segment 3:  -10  
   5193 .. 5199  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5169 .. 5199  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
4889 .. 4913  =  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
4889 .. 4913  =  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).
Strep-Tag II
4987 .. 5010  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an
engineered form of streptavidin
Strep-Tag II
4987 .. 5010  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an
engineered form of streptavidin
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
4963 .. 4980  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4963 .. 4980  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
4944 .. 4949  =  6 bp
ribosome binding site
RBS
4944 .. 4949  =  6 bp
ribosome binding site
ATG
4957 .. 4959  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4957 .. 4959  =  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