pcDNA6.2 nGeneBLAzer-DEST

Gateway® destination vector for in vivo or in vitro detection of mammalian gene expression using an N-terminal β-lactamase reporter.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspcDNA6.2 nGeneBLAzer-DEST
SgrDI (7562) PciI (5751) BspQI - SapI (5635) PfoI (5175) BsgI (4871) PpuMI (4843) BbsI (4769) BclI * (4606) AvrII (4555) StuI (4554) CsiI - SexAI * (4322) MfeI (161) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) NotI (1838) BspEI (2154) EcoRI (2158) BsmBI - Esp3I (2382) PflMI * (2390) BssHII (2652) BbvCI (2883) SrfI (3029) BmgBI (3063) BstXI (3146) BfuAI - BspMI (3273) PstI (3284) AgeI (3469) PmeI (3487) pcDNA™6.2/nGeneBLAzer™-DEST 7564 bp
SgrDI  (7562)
1 site		 C G T C G A C G G C A G C T G C
PciI  (5751)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (5635)
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  (5635)
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.
PfoI  (5175)
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.
BsgI  (4871)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PpuMI  (4843)
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.
BbsI  (4769)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BclI  (4606)
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.
AvrII  (4555)
1 site		 C C T A G G G G A T C C
StuI  (4554)
1 site		 A G G C C T T C C G G A
CsiI  (4322)
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  (4322)
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.
MfeI  (161)
1 site		 C A A T T G G T T A A C
SpeI  (249)
1 site		 A C T A G T T G A T C A
NdeI  (484)
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.
SnaBI  (590)
1 site		 T A C G T A A T G C A T
Eco53kI  (816)
1 site		 G A G C T C C T C G A G
SacI  (818)
1 site		 G A G C T C C T C G A G
NotI  (1838)
1 site		 G C G G C C G C C G C C G G C G
BspEI  (2154)
1 site		 T C C G G A A G G C C T
EcoRI  (2158)
1 site		 G A A T T C C T T A A G
BsmBI  (2382)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2382)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PflMI  (2390)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BssHII  (2652)
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.
BbvCI  (2883)
1 site		 C C T C A G C G G A G T C G
SrfI  (3029)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (3063)
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.
BstXI  (3146)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BfuAI  (3273)
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  (3273)
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.
PstI  (3284)
1 site		 C T G C A G G A C G T C
AgeI  (3469)
1 site		 A C C G G T T G G C C A
PmeI  (3487)
1 site		 G T T T A A A C C A A A T T T G
AmpR
6568 .. 7428  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6568 .. 7359  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6568 .. 7428  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7360 .. 7428  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6568 .. 7428  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
bla(M)
911 .. 1702  =  792 bp
264 amino acids  =  29.1 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
bla(M)
911 .. 1702  =  792 bp
264 amino acids  =  29.1 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
CmR
1945 .. 2604  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1945 .. 2604  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5812 .. 6397  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5812 .. 6397  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3799 .. 4227  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3799 .. 4227  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4684 .. 5082  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4684 .. 5082  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4241 .. 4570  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4241 .. 4570  =  330 bp
SV40 enhancer and early promoter
ccdB
2946 .. 3251  =  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
2946 .. 3251  =  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.
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
attR1
1712 .. 1836  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1712 .. 1836  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3292 .. 3416  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3292 .. 3416  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5240 .. 5361  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5240 .. 5361  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7429 .. 7533  =  105 bp
AmpR promoter
7429 .. 7533  =  105 bp
HSV TK poly(A) signal
3549 .. 3597  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3549 .. 3597  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
4618 .. 4665  =  48 bp
synthetic bacterial promoter
EM7 promoter
4618 .. 4665  =  48 bp
synthetic bacterial promoter
V5 tag
3424 .. 3465  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3424 .. 3465  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
1861 .. 1891  =  31 bp
3 segments
   Segment 1:  -35  
   1861 .. 1866  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1861 .. 1891  =  31 bp
3 segments
   Segment 2:  
   1867 .. 1884  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1861 .. 1891  =  31 bp
3 segments
   Segment 3:  -10  
   1885 .. 1891  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1861 .. 1891  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4421 .. 4556  =  136 bp
SV40 origin of replication
SV40 ori
4421 .. 4556  =  136 bp
SV40 origin of replication
ORF:  1945 .. 2604  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4684 .. 5082  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  911 .. 1747  =  837 bp
ORF:  278 amino acids  =  30.7 kDa
ORF:  6698 .. 6964  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2946 .. 3251  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  1309 .. 1575  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4630 .. 4986  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  6568 .. 7428  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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