pCOLA-2-DEST

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

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsGateway® Cloning VectorspCOLA-2-DEST
EcoO109I (5094) BlpI (5067) AvrII (5049) BstBI (5030) SalI (4876) PstI (4874) BfuAI - BspMI (4863) BsaBI * (4632) SrfI (4619) AlwNI (4527) BbvCI (4473) BstZ17I (4283) ScaI (4165) PasI (3980) EcoRI (3748) BspEI (3744) EagI - NotI (3428) 6xHis 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-2-DEST™ 5131 bp
EcoO109I  (5094)
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  (5067)
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  (5049)
1 site		 C C T A G G G G A T C C
BstBI  (5030)
1 site		 T T C G A A A A G C T T
SalI  (4876)
1 site		 G T C G A C C A G C T G
PstI  (4874)
1 site		 C T G C A G G A C G T C
BfuAI  (4863)
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  (4863)
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  (4632)
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  (4619)
1 site		 G C C C G G G C C G G G C C C G
AlwNI  (4527)
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  (4473)
1 site		 C C T C A G C G G A G T C G
BstZ17I  (4283)
1 site		 G T A T A C C A T A T G
ScaI  (4165)
1 site		 A G T A C T T C A T G A
PasI  (3980)
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  (3748)
1 site		 G A A T T C C T T A A G
BspEI  (3744)
1 site		 T C C G G A A G G C C T
EagI  (3428)
1 site		 C G G C C G G C C G G C
NotI  (3428)
1 site		 G C G G C C G C C G C C G G C G
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
3535 .. 4194  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3535 .. 4194  =  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
4536 .. 4841  =  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
4536 .. 4841  =  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
3302 .. 3426  =  125 bp
recombination site for the Gateway® LR reaction
attR1
3302 .. 3426  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4882 .. 5006  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4882 .. 5006  =  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
5078 .. 5125  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
5078 .. 5125  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
3451 .. 3481  =  31 bp
3 segments
   Segment 1:  -35  
   3451 .. 3456  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3451 .. 3481  =  31 bp
3 segments
   Segment 2:  
   3457 .. 3474  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3451 .. 3481  =  31 bp
3 segments
   Segment 3:  -10  
   3475 .. 3481  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3451 .. 3481  =  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).
Strep-Tag II
5014 .. 5037  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
Strep-Tag II
5014 .. 5037  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
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
6xHis
3281 .. 3298  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3281 .. 3298  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 promoter
3188 .. 3206  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3188 .. 3206  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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:  3535 .. 4194  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4536 .. 4841  =  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-2-DEST.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

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