pENTR GeneBLAzer

Gateway® entry clone to be used as a control for fluorescence detection of β-lactamase with the GeneBLAzer® system.

Sequence Author: Thermo Fisher (Invitrogen)

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PciI (3333) DrdI (3231) BciVI (3135) PspFI (3033) BseYI (3029) AlwNI (2924) PflMI (2421) Bpu10I (2175) BsmBI - Esp3I (2174) AsiSI (2158) SspI (2083) EcoNI (2070) NruI (1815) NspI (3337) BspQI - SapI (111) rrnB T2 terminator BbsI (437) HincII - HpaI (501) AflII (554) AvaI - BsoBI (560) PspOMI (563) EcoO109I (564) ApaI (567) BtgI - NcoI - StyI (669) XmnI (792) DraI (814) ScaI (911) FspI (1169) NmeAIII (1245) BglI (1274) BpmI (1323) BsaI (1326) AccI (1463) EcoRV (1576) pENTR™/GeneBLAzer® 3339 bp
PciI  (3333)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (3231)
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.
BciVI  (3135)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PspFI  (3033)
1 site		 C C C A G C G G G T C G
BseYI  (3029)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2924)
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.
PflMI  (2421)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
Bpu10I  (2175)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsmBI  (2174)
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  (2174)
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.
AsiSI  (2158)
1 site		 G C G A T C G C C G C T A G C G
SspI  (2083)
1 site		 A A T A T T T T A T A A
EcoNI  (2070)
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.
NruI  (1815)
1 site		 T C G C G A A G C G C T
NspI  (3337)
1 site		 R C A T G Y Y G T A C R
BspQI  (111)
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  (111)
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.
BbsI  (437)
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.
HincII  (501)
1 site		 G T Y R A C C A R Y T G
HpaI  (501)
1 site		 G T T A A C C A A T T G
AflII  (554)
1 site		 C T T A A G G A A T T C
AvaI  (560)
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  (560)
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.
PspOMI  (563)
1 site		 G G G C C C C C C G G G
EcoO109I  (564)
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.
ApaI  (567)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BtgI  (669)
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  (669)
1 site		 C C A T G G G G T A C C
StyI  (669)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
XmnI  (792)
1 site		 G A A N N N N T T C C T T N N N N A A G
DraI  (814)
1 site		 T T T A A A A A A T T T
ScaI  (911)
1 site		 A G T A C T T C A T G A
FspI  (1169)
1 site		 T G C G C A A C G C G T
NmeAIII  (1245)
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).
BglI  (1274)
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.
BpmI  (1323)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsaI  (1326)
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.
AccI  (1463)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
EcoRV  (1576)
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.
KanR
1733 .. 2542  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1733 .. 2542  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
bla(M)
671 .. 1465  =  795 bp
264 amino acids  =  29.1 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
bla(M)
671 .. 1465  =  795 bp
264 amino acids  =  29.1 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
ori
2689 .. 3277  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2689 .. 3277  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
attL1
569 .. 667  =  99 bp
recombination site for the Gateway® LR reaction
attL1
569 .. 667  =  99 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
attL2
1464 .. 1563  =  100 bp
recombination site for the Gateway® LR reaction
attL2
1464 .. 1563  =  100 bp
recombination site for the Gateway® LR reaction
Kozak sequence
668 .. 674  =  7 bp
translation initiation signal for mammalian cells
Kozak sequence
668 .. 674  =  7 bp
translation initiation signal for mammalian cells
ORF:  641 .. 1465  =  825 bp
ORF:  274 amino acids  =  30.1 kDa
ORF:  1733 .. 2542  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
ORF:  394 .. 672  =  279 bp
ORF:  92 amino acids  =  10.2 kDa
ORF:  1069 .. 1335  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pENTR GeneBLAzer.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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