pSCREEN-iT lacZ-DEST

Gateway® destination vector that allows assessment of the potency of an RNAi knockdown reagent using a β-gal reporter assay.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspSCREEN-iT lacZ-DEST
SgrDI (8700) AhdI (7779) BspQI - SapI (6770) PsiI (6363) BstAPI (5959) EcoO109I (5956) PmeI (5693) PstI (5541) BmgBI (5320) SmaI - SrfI (5286) TspMI - XmaI (5284) BbvCI (5140) BspEI (4411) MfeI (161) NruI (208) SpeI (249) SnaBI (590) Bsu36I (1114) BspDI - ClaI (1714) EcoRV (2003) BclI * (2236) MauBI (2388) AfeI (2725) BsiWI (3661) PfoI * (3870) BlpI (3903) PaeR7I - PspXI - XhoI (3955) pSCREEN-iT™/lacZ-DEST 8702 bp
SgrDI  (8700)
1 site		 C G T C G A C G G C A G C T G C
AhdI  (7779)
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.
BspQI  (6770)
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  (6770)
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.
PsiI  (6363)
1 site		 T T A T A A A A T A T T
BstAPI  (5959)
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.
EcoO109I  (5956)
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.
PmeI  (5693)
1 site		 G T T T A A A C C A A A T T T G
PstI  (5541)
1 site		 C T G C A G G A C G T C
BmgBI  (5320)
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  (5286)
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  (5286)
1 site		 G C C C G G G C C G G G C C C G
TspMI  (5284)
1 site		 C C C G G G G G G C C C
XmaI  (5284)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BbvCI  (5140)
1 site		 C C T C A G C G G A G T C G
BspEI  (4411)
1 site		 T C C G G A A G G C C T
MfeI  (161)
1 site		 C A A T T G G T T A A C
NruI  (208)
1 site		 T C G C G A A G C G C T
SpeI  (249)
1 site		 A C T A G T T G A T C A
SnaBI  (590)
1 site		 T A C G T A A T G C A T
Bsu36I  (1114)
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.
BspDI  (1714)
1 site		 A T C G A T T A G C T A
ClaI  (1714)
1 site		 A T C G A T T A G C T A
EcoRV  (2003)
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.
BclI  (2236)
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.
MauBI  (2388)
1 site		 C G C G C G C G G C G C G C G C
AfeI  (2725)
1 site		 A G C G C T T C G C G A
BsiWI  (3661)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
PfoI  (3870)
1 site		 T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BlpI  (3903)
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.
PaeR7I  (3955)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3955)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (3955)
1 site		 C T C G A G G A G C T C
lacZ
903 .. 3947  =  3045 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
903 .. 3947  =  3045 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
AmpR
7706 .. 8566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7706 .. 8497  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7706 .. 8566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8498 .. 8566  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7706 .. 8566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
4202 .. 4861  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
4202 .. 4861  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
6947 .. 7535  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6947 .. 7535  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6005 .. 6433  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6005 .. 6433  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
ccdB
5203 .. 5508  =  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
5203 .. 5508  =  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
3969 .. 4093  =  125 bp
recombination site for the Gateway® LR reaction
attR1
3969 .. 4093  =  125 bp
recombination site for the Gateway® LR reaction
attR2
5549 .. 5673  =  125 bp
recombination site for the Gateway® LR reaction
attR2
5549 .. 5673  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
8567 .. 8671  =  105 bp
AmpR promoter
8567 .. 8671  =  105 bp
HSV TK poly(A) signal
5755 .. 5803  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
5755 .. 5803  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
lac UV5 promoter
4118 .. 4148  =  31 bp
3 segments
   Segment 1:  -35  
   4118 .. 4123  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
4118 .. 4148  =  31 bp
3 segments
   Segment 2:  
   4124 .. 4141  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
4118 .. 4148  =  31 bp
3 segments
   Segment 3:  -10  
   4142 .. 4148  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
4118 .. 4148  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  4202 .. 4861  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  774 .. 4004  =  3231 bp
ORF:  1076 amino acids  =  122.7 kDa
ORF:  7706 .. 8566  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pSCREEN-iT lacZ-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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