pCOLA-3-DEST

Gateway® Nova bacterial destination vector with a ColA origin for expressing proteins tagged at the N-terminus with Strep-Tag II.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pCOLA-3-DEST
EcoO109I (5097) BlpI (5070) AvrII (5052) DraIII (5043) 6xHis SalI (4891) PstI (4889) BfuAI - BspMI (4878) BsaBI * (4647) SrfI (4634) AlwNI (4542) BbvCI (4488) BstZ17I (4298) ScaI (4180) PasI (3995) EcoRI (3763) BspEI (3759) EagI - NotI (3443) BstBI (3300) Strep-Tag II BtgI - NcoI (3273) RBS T7 promoter PfoI (3141) BstAPI (3033) BclI * (2695) BstEII (2527) Bsu36I (2) AgeI (51) DrdI - PflFI - Tth111I (122) BsaI (130) XmnI (218) BspDI - ClaI (914) NruI (950) NheI - NspI - SphI (1139) BmtI (1143) EcoRV (1565) CsiI - SexAI * (1648) PvuI (1740) XbaI (1789) AcuI (1849) KasI (2070) NarI * (2071) SfoI (2072) PluTI (2074) HpaI (2207) PspOMI (2502) ApaI (2506) NmeAIII (2509) pCOLA-3-DEST™ 5134 bp
EcoO109I  (5097)
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.
BlpI  (5070)
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.
AvrII  (5052)
1 site		 C C T A G G G G A T C C
DraIII  (5043)
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.
SalI  (4891)
1 site		 G T C G A C C A G C T G
PstI  (4889)
1 site		 C T G C A G G A C G T C
BfuAI  (4878)
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  (4878)
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.
BsaBI  (4647)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
SrfI  (4634)
1 site		 G C C C G G G C C G G G C C C G
AlwNI  (4542)
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.
BbvCI  (4488)
1 site		 C C T C A G C G G A G T C G
BstZ17I  (4298)
1 site		 G T A T A C C A T A T G
ScaI  (4180)
1 site		 A G T A C T T C A T G A
PasI  (3995)
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  (3763)
1 site		 G A A T T C C T T A A G
BspEI  (3759)
1 site		 T C C G G A A G G C C T
EagI  (3443)
1 site		 C G G C C G G C C G G C
NotI  (3443)
1 site		 G C G G C C G C C G C C G G C G
BstBI  (3300)
1 site		 T T C G A A A A G C T T
BtgI  (3273)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
NcoI  (3273)
1 site		 C C A T G G G G T A C C
PfoI  (3141)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
BstAPI  (3033)
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  (2695)
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  (2527)
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.
Bsu36I  (2)
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.
AgeI  (51)
1 site		 A C C G G T T G G C C A
DrdI  (122)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
PflFI  (122)
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  (122)
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.
BsaI  (130)
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.
XmnI  (218)
1 site		 G A A N N N N T T C C T T N N N N A A G
BspDI  (914)
1 site		 A T C G A T T A G C T A
ClaI  (914)
1 site		 A T C G A T T A G C T A
NruI  (950)
1 site		 T C G C G A A G C G C T
NheI  (1139)
1 site		 G C T A G C C G A T C G
NspI  (1139)
1 site		 R C A T G Y Y G T A C R
SphI  (1139)
1 site		 G C A T G C C G T A C G
BmtI  (1143)
1 site		 G C T A G C C G A T C G
EcoRV  (1565)
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.
CsiI  (1648)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (1648)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
PvuI  (1740)
1 site		 C G A T C G G C T A G C
XbaI  (1789)
1 site		 T C T A G A A G A T C T
AcuI  (1849)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
KasI  (2070)
1 site		 G G C G C C C C G C G G
NarI  (2071)
1 site		 G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2072)
1 site		 G G C G C C C C G C G G
PluTI  (2074)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
HpaI  (2207)
1 site		 G T T A A C C A A T T G
PspOMI  (2502)
1 site		 G G G C C C C C C G G G
ApaI  (2506)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
NmeAIII  (2509)
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).
lacI
1982 .. 3064  =  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
1982 .. 3064  =  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
224 .. 1039  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
224 .. 1039  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
CmR
3550 .. 4209  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3550 .. 4209  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ColA ori
1149 .. 1784  =  636 bp
Plasmids containing the ColA origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
ColA ori
1149 .. 1784  =  636 bp
Plasmids containing the ColA origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
ccdB
4551 .. 4856  =  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
4551 .. 4856  =  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.
attR1
3317 .. 3441  =  125 bp
recombination site for the Gateway® LR reaction
attR1
3317 .. 3441  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4897 .. 5021  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4897 .. 5021  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
1040 .. 1131  =  92 bp
AmpR promoter
1040 .. 1131  =  92 bp
lacI promoter
3065 .. 3142  =  78 bp
lacI promoter
3065 .. 3142  =  78 bp
T7 terminator
5081 .. 5128  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
5081 .. 5128  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ATG
3275 .. 3277  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3275 .. 3277  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Strep-Tag II
3284 .. 3307  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
Strep-Tag II
3284 .. 3307  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
lac UV5 promoter
3466 .. 3496  =  31 bp
3 segments
   Segment 1:  -35  
   3466 .. 3471  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3466 .. 3496  =  31 bp
3 segments
   Segment 2:  
   3472 .. 3489  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3466 .. 3496  =  31 bp
3 segments
   Segment 3:  -10  
   3490 .. 3496  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3466 .. 3496  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
3207 .. 3231  =  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
3207 .. 3231  =  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).
T7 promoter
3188 .. 3206  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3188 .. 3206  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
5023 .. 5040  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5023 .. 5040  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
3262 .. 3267  =  6 bp
ribosome binding site
RBS
3262 .. 3267  =  6 bp
ribosome binding site
ORF:  1318 .. 1554  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  1948 .. 2199  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3550 .. 4209  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4551 .. 4856  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  224 .. 1039  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  1982 .. 2941  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1962 .. 2225  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2814 .. 3179  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
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Download pCOLA-3-DEST.dna file

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Individual Sequences & Maps

pACYCDuet-1
pET-24a(+)
pET-3a
pET-51b(+) Ek_LIC (linearized)
pCDF-1b
pET-24b(+)
pET-3b
pET-52b(+)
pCDF-2 Ek_LIC
pET-24c(+)
pET-3c
pET-52b(+) 3C_LIC
pCDF-2 Ek_LIC (linearized)
pET-24d(+)
pET-3d
pET-52b(+) 3C_LIC (linearized)
pCDFDuet-1
pET-25b(+)
pET-40b(+)
pET-53-DEST
pCOLA-2-DEST
pET-26b(+)
pET-41 Ek_LIC
pET-54-DEST
pCOLA-3-DEST
pET-27b(+)
pET-41 Ek_LIC (linearized)
pET-55-DEST
pCOLADuet-1
pET-28a(+)
pET-41a(+)
pET-56-DEST
pET-11a
pET-28b(+)
pET-41b(+)
pET-57-DEST
pET-11b
pET-28c(+)
pET-41c(+)
pET-58-DEST
pET-11c
pET-29a(+)
pET-42a(+)
pET-59-DEST
pET-11d
pET-29b(+)
pET-42b(+)
pET-60-DEST
pET-14b
pET-29c(+)
pET-42c(+)
pET-61-DEST
pET-15b
pET-30 Ek_LIC
pET-43.1 Ek_LIC
pET-62-DEST
pET-16b
pET-30 Ek_LIC (linearized)
pET-43.1 Ek_LIC (linearized)
pET-9a
pET-17b
pET-30 Xa_LIC
pET-43.1a(+)
pET-9b
pET-19b
pET-30 Xa_LIC (linearized)
pET-43.1b(+)
pET-9c
pET-20b(+)
pET-30a(+)
pET-43.1c(+)
pET-9d
pET-21(+)
pET-30b(+)
pET-44a(+)
pETBlue-1
pET-21a(+)
pET-30c(+)
pET-44b(+)
pETBlue-2
pET-21b(+)
pET-31b(+)
pET-44c(+)
pETcoco-1
pET-21c(+)
pET-32 Ek_LIC
pET-45b(+)
pETcoco-2
pET-21d(+)
pET-32 Ek_LIC (linearized)
pET-46 Ek_LIC
pETDuet-1
pET-22b(+)
pET-32 Xa_LIC
pET-46 Ek_LIC (linearized)
pLacI
pET-23(+)
pET-32 Xa_LIC (linearized)
pET-47b(+)
pLysE
pET-23a(+)
pET-32a(+)
pET-48b(+)
pLysS
pET-23b(+)
pET-32b(+)
pET-49b(+)
pRSF-1b
pET-23c(+)
pET-32c(+)
pET-50b(+)
pRSF-2 Ek_LIC
pET-23d(+)
pET-33b(+)
pET-51b(+)
pRSF-2 Ek_LIC (linearized)
pET-24(+)
pET-39b(+)
pET-51b(+) Ek_LIC
pRSFDuet-1

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