pHELLSGATE

High-throughput binary Gateway® vector for silencing genes in plants using intron-containing hairpin RNA.
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HomePlasmidsGateway® Cloning VectorspHELLSGATE
AgeI (18,257) XbaI (17,913) XbaI (16,296) HindIII (16,284) BspDI - ClaI (16,279) PacI (16,168) KpnI (14,653) Acc65I (14,649) AbsI - XhoI (14,637) XhoI (13,020) StuI (12,234) SacI (11,646) Eco53kI (11,644) PmeI (11,220) BsiWI (11,055) FseI (10,595) Bsu36I (330) RsrII (1119) AflII (1923) MauBI (2118) LB T-DNA repeat AleI (5445) SexAI * (5533) FspAI (6101) NruI (6598) pHELLSGATE 18,691 bp
AgeI  (18,257)
1 site		 A C C G G T T G G C C A
XbaI  (17,913)
2 sites		 T C T A G A A G A T C T
XbaI  (16,296)
2 sites		 T C T A G A A G A T C T
HindIII  (16,284)
1 site		 A A G C T T T T C G A A
BspDI  (16,279)
1 site		 A T C G A T T A G C T A
ClaI  (16,279)
1 site		 A T C G A T T A G C T A
PacI  (16,168)
1 site		 T T A A T T A A A A T T A A T T
KpnI  (14,653)
1 site		 G G T A C C C C A T G G
Acc65I  (14,649)
1 site		 G G T A C C C C A T G G
AbsI  (14,637)
1 site		 C C T C G A G G G G A G C T C C
XhoI  (14,637)
2 sites		 C T C G A G G A G C T C
XhoI  (13,020)
2 sites		 C T C G A G G A G C T C
StuI  (12,234)
1 site		 A G G C C T T C C G G A
SacI  (11,646)
1 site		 G A G C T C C T C G A G
Eco53kI  (11,644)
1 site		 G A G C T C C T C G A G
PmeI  (11,220)
1 site		 G T T T A A A C C A A A T T T G
BsiWI  (11,055)
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.
FseI  (10,595)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
Bsu36I  (330)
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.
RsrII  (1119)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
AflII  (1923)
1 site		 C T T A A G G A A T T C
MauBI  (2118)
1 site		 C G C G C G C G G C G C G C G C
AleI  (5445)
1 site		 C A C N N N N G T G G T G N N N N C A C
SexAI  (5533)
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.
FspAI  (6101)
1 site		 R T G C G C A Y Y A C G C G T R
NruI  (6598)
1 site		 T C G C G A A G C G C T
PDK intron
14,662 .. 16,258  =  1597 bp
modified pyruvate orthophosphate dikinase intron from Flaveria trinervia
PDK intron
14,662 .. 16,258  =  1597 bp
modified pyruvate orthophosphate dikinase intron from Flaveria trinervia
trfA
4407 .. 5555  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
trfA
4407 .. 5555  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
KanR
448 .. 1269  =  822 bp
273 amino acids  =  29.9 kDa
3 segments
   Segment 1:  nos  
   448 .. 489  =  42 bp
   14 amino acids  =  1.4 kDa
Product: fusion between an N-terminal peptide of nopaline synthase and Tn5 aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
448 .. 1269  =  822 bp
273 amino acids  =  29.9 kDa
3 segments
   Segment 2:  
   490 .. 498  =  9 bp
   3 amino acids  =  443.5 Da
Product: fusion between an N-terminal peptide of nopaline synthase and Tn5 aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
448 .. 1269  =  822 bp
273 amino acids  =  29.9 kDa
3 segments
   Segment 3:  nptII  
   499 .. 1269  =  771 bp
   256 amino acids  =  28.1 kDa
Product: fusion between an N-terminal peptide of nopaline synthase and Tn5 aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
448 .. 1269  =  822 bp
273 amino acids  =  29.9 kDa
3 segments
Product: fusion between an N-terminal peptide of nopaline synthase and Tn5 aminoglycoside phosphotransferase
confers resistance to kanamycin
SmR
8600 .. 9388  =  789 bp
262 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
SmR
8600 .. 9388  =  789 bp
262 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
IS1
3015 .. 3782  =  768 bp
prokaryotic transposable element
IS1
3015 .. 3782  =  768 bp
prokaryotic transposable element
OCS terminator
17,922 .. 18,628  =  707 bp
octopine synthase terminator
OCS terminator
17,922 .. 18,628  =  707 bp
octopine synthase terminator
oriV
10,071 .. 10,684  =  614 bp
origin of replication for the bacterial F plasmid
oriV
10,071 .. 10,684  =  614 bp
origin of replication for the bacterial F plasmid
ori
6915 .. 7503  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6915 .. 7503  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CaMV 35S promoter
12,672 .. 13,017  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
12,672 .. 13,017  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
ccdB
13,675 .. 13,980  =  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
13,675 .. 13,980  =  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
16,855 .. 17,160  =  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
16,855 .. 17,160  =  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.
NOS terminator
1894 .. 2146  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
1894 .. 2146  =  253 bp
nopaline synthase terminator and poly(A) signal
attP1
13,048 .. 13,279  =  232 bp
recombination site for the Gateway® BP reaction
attP1
13,048 .. 13,279  =  232 bp
recombination site for the Gateway® BP reaction
attP2
14,387 .. 14,618  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™221 version)
attP2
14,387 .. 14,618  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™221 version)
attP2
16,320 .. 16,551  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™221 version)
attP2
16,320 .. 16,551  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™221 version)
attP1
17,659 .. 17,890  =  232 bp
recombination site for the Gateway® BP reaction
attP1
17,659 .. 17,890  =  232 bp
recombination site for the Gateway® BP reaction
NOS promoter
264 .. 447  =  184 bp
Function: NOS promoter
nopaline synthase promoter
NOS promoter
264 .. 447  =  184 bp
Function: NOS promoter
nopaline synthase promoter
oriT
2846 .. 2955  =  110 bp
incP origin of transfer
oriT
2846 .. 2955  =  110 bp
incP origin of transfer
LB T-DNA repeat
2268 .. 2292  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
2268 .. 2292  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
11,182 .. 11,206  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
11,182 .. 11,206  =  25 bp
right border repeat from nopaline C58 T-DNA
CmR
15,002 .. 15,661  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
15,002 .. 15,661  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ORF:  448 .. 1269  =  822 bp
ORF:  273 amino acids  =  29.9 kDa
ORF:  4996 .. 5268  =  273 bp
ORF:  90 amino acids  =  10.2 kDa
ORF:  12,175 .. 12,438  =  264 bp
ORF:  87 amino acids  =  10.4 kDa
ORF:  12,490 .. 12,918  =  429 bp
ORF:  142 amino acids  =  15.9 kDa
ORF:  647 .. 1033  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1637 .. 1873  =  237 bp
ORF:  78 amino acids  =  8.5 kDa
ORF:  3869 .. 4135  =  267 bp
ORF:  88 amino acids  =  9.7 kDa
ORF:  5135 .. 5530  =  396 bp
ORF:  131 amino acids  =  14.1 kDa
ORF:  5810 .. 6709  =  900 bp
ORF:  299 amino acids  =  31.8 kDa
ORF:  8018 .. 8542  =  525 bp
ORF:  174 amino acids  =  19.7 kDa
ORF:  8600 .. 9388  =  789 bp
ORF:  262 amino acids  =  29.2 kDa
ORF:  9464 .. 9961  =  498 bp
ORF:  165 amino acids  =  18.0 kDa
ORF:  15,002 .. 15,661  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3264 .. 3767  =  504 bp
ORF:  167 amino acids  =  19.6 kDa
ORF:  4407 .. 5555  =  1149 bp
ORF:  382 amino acids  =  43.8 kDa
ORF:  12,096 .. 12,500  =  405 bp
ORF:  134 amino acids  =  15.5 kDa
ORF:  3443 .. 3733  =  291 bp
ORF:  96 amino acids  =  11.0 kDa
ORF:  4349 .. 4936  =  588 bp
ORF:  195 amino acids  =  20.7 kDa
ORF:  5705 .. 5971  =  267 bp
ORF:  88 amino acids  =  9.8 kDa
ORF:  8639 .. 9022  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  784 .. 1320  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  8200 .. 8451  =  252 bp
ORF:  83 amino acids  =  9.3 kDa
ORF:  8632 .. 8889  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  8899 .. 9177  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  10,540 .. 10,794  =  255 bp
ORF:  84 amino acids  =  10.0 kDa
ORF:  13,675 .. 13,980  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  16,855 .. 17,160  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2190 .. 2414  =  225 bp
ORF:  74 amino acids  =  8.2 kDa
ORF:  5148 .. 5543  =  396 bp
ORF:  131 amino acids  =  14.7 kDa
ORF:  7674 .. 7910  =  237 bp
ORF:  78 amino acids  =  8.2 kDa
ORF:  9462 .. 9689  =  228 bp
ORF:  75 amino acids  =  8.7 kDa
ORF:  12,123 .. 12,491  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
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Download pHELLSGATE.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

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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