pHELLSGATE 12

High-throughput binary Gateway® destination vector with two introns, for silencing genes in plants using intron-containing hairpin RNA.
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HomePlasmidsPlant VectorspHELLSGATE 12
AgeI (17,247) XbaI (16,903) XbaI (15,484) BspDI - ClaI (15,262) PacI (15,151) PDK intron KpnI (14,465) Acc65I (14,461) EcoRI (14,455) XhoI (14,449) XhoI (13,020) ScaI (12,386) 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 BspEI (3460) AleI (5445) SexAI * (5533) FspAI (6101) NruI (6598) pHELLSGATE 12 17,681 bp
AgeI  (17,247)
1 site		 A C C G G T T G G C C A
XbaI  (16,903)
2 sites		 T C T A G A A G A T C T
XbaI  (15,484)
2 sites		 T C T A G A A G A T C T
BspDI  (15,262)
1 site		 A T C G A T T A G C T A
ClaI  (15,262)
1 site		 A T C G A T T A G C T A
PacI  (15,151)
1 site		 T T A A T T A A A A T T A A T T
KpnI  (14,465)
1 site		 G G T A C C C C A T G G
Acc65I  (14,461)
1 site		 G G T A C C C C A T G G
EcoRI  (14,455)
1 site		 G A A T T C C T T A A G
XhoI  (14,449)
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
ScaI  (12,386)
1 site		 A G T A C T T C A T G A
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
BspEI  (3460)
1 site		 T C C G G A A G G C C T
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
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
PDK intron
14,475 .. 15,241  =  767 bp
pyruvate orthophosphate dikinase intron from Flaveria trinervia
PDK intron
14,475 .. 15,241  =  767 bp
pyruvate orthophosphate dikinase intron from Flaveria trinervia
OCS terminator
16,912 .. 17,618  =  707 bp
octopine synthase terminator
OCS terminator
16,912 .. 17,618  =  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
13,579 .. 13,884  =  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
13,579 .. 13,884  =  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
15,942 .. 16,247  =  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
15,942 .. 16,247  =  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
cat1 intron
15,276 .. 15,465  =  190 bp
castor bean catalase intron, modified
cat1 intron
15,276 .. 15,465  =  190 bp
castor bean catalase intron, modified
NOS promoter
264 .. 447  =  184 bp
Function: NOS promoter
nopaline synthase promoter
NOS promoter
264 .. 447  =  184 bp
Function: NOS promoter
nopaline synthase promoter
attR1
13,026 .. 13,150  =  125 bp
recombination site for the Gateway® LR reaction
attR1
13,026 .. 13,150  =  125 bp
recombination site for the Gateway® LR reaction
attR2
14,324 .. 14,448  =  125 bp
recombination site for the Gateway® LR reaction
attR2
14,324 .. 14,448  =  125 bp
recombination site for the Gateway® LR reaction
attR2
15,489 .. 15,613  =  125 bp
recombination site for the Gateway® LR reaction
attR2
15,489 .. 15,613  =  125 bp
recombination site for the Gateway® LR reaction
attR1
16,779 .. 16,903  =  125 bp
recombination site for the Gateway® LR reaction
attR1
16,779 .. 16,903  =  125 bp
recombination site for the Gateway® LR reaction
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
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:  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:  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
ORF:  15,942 .. 16,247  =  306 bp
ORF:  101 amino acids  =  11.7 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:  13,579 .. 13,884  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
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Individual Sequences & Maps

pBI121
pCAMBIA1381Xa
pEarleyGate 201
pPZP100
pBI221
pCAMBIA1381Xb
pEarleyGate 202
pPZP101
pBin19
pCAMBIA1381Xc
pEarleyGate 203
pPZP102
pBINPLUS
pCAMBIA1381Z
pEarleyGate 204
pPZP111
pCAMBIA0105.1R
pCAMBIA1390
pEarleyGate 205
pPZP112
pCAMBIA0305.1
pCAMBIA1391
pEarleyGate 301
pPZP121
pCAMBIA0305.2
pCAMBIA1391Xa
pEarleyGate 302
pPZP122
pCAMBIA0380
pCAMBIA1391Xb
pEarleyGate 303
pPZP200
pCAMBIA0390
pCAMBIA1391Xc
pEarleyGate 304
pPZP201
pCAMBIA1105.1
pCAMBIA1391Z
pFGC5941
pPZP202
pCAMBIA1105.1R
pCAMBIA2200
pGA643
pPZP211
pCAMBIA1200
pCAMBIA2201
pGreen
pPZP212
pCAMBIA1201
pCAMBIA2300
pGreen 0029
pPZP221
pCAMBIA1281Z
pCAMBIA2301
pGreenII
pPZP222
pCAMBIA1291Z
pCAMBIA3200
pGreenII 0049
pRI 101-AN
pCAMBIA1300
pCAMBIA3201
pGreenII 0179
pRI 101-ON
pCAMBIA1301
pCAMBIA3300
pGreenII 0229
pRI 201-AN
pCAMBIA1302
pCAMBIA3301
pGreenII 0579
pRI 201-ON
pCAMBIA1303
pCAMBIA5105
pHANNIBAL
pRI 909
pCAMBIA1304
pEarleyGate 100
pHELLSGATE
pRI 910
pCAMBIA1305.1
pEarleyGate 101
pHELLSGATE 12
pSB1
pCAMBIA1305.2
pEarleyGate 102
pHELLSGATE 4
pSB11
pCAMBIA1380
pEarleyGate 103
pHELLSGATE 8
pSoup
pCAMBIA1381
pEarleyGate 104
pKANNIBAL

Plasmid Sets

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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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