pEarleyGate 204

Gateway®-compatible plant transformation vector with an N-terminal AcV5 tag.

Sequence Author: Pikaard Lab

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HomePlasmidsPlant VectorspEarleyGate 204
LB T-DNA repeat BclI * (11,543) PsiI (11,412) BspHI (11,086) EcoRV (10,663) PluTI (9404) SfoI (9402) NarI (9401) KasI (9400) EcoNI (8928) BmtI (8619) NheI (8615) BsiWI (8499) PmeI (5487) AseI (92) Acc65I (407) KpnI (411) FspAI (521) PaqCI (585) AfeI (758) BstBI (1770) StuI (1868) PshAI (2467) PaeR7I - PspXI - XhoI (2654) TMV Ω ATG AcV5 tag attR1 lac UV5 promoter BspEI (3212) NcoI (3517) BbvCI (3941) SrfI (4087) BstXI (4204) AvrII (4476) PacI (4523) SpeI (4535) MfeI (4839) BstEII (5111) AhdI (5241) SbfI (5266) HindIII (5274) PvuI (5400) pEarleyGate 204 11,780 bp
BclI  (11,543)
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.
PsiI  (11,412)
1 site		 T T A T A A A A T A T T
BspHI  (11,086)
1 site		 T C A T G A A G T A C T
EcoRV  (10,663)
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.
PluTI  (9404)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (9402)
1 site		 G G C G C C C C G C G G
NarI  (9401)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (9400)
1 site		 G G C G C C C C G C G G
EcoNI  (8928)
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.
BmtI  (8619)
1 site		 G C T A G C C G A T C G
NheI  (8615)
1 site		 G C T A G C C G A T C G
BsiWI  (8499)
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.
PmeI  (5487)
1 site		 G T T T A A A C C A A A T T T G
AseI  (92)
1 site		 A T T A A T T A A T T A
Acc65I  (407)
1 site		 G G T A C C C C A T G G
KpnI  (411)
1 site		 G G T A C C C C A T G G
FspAI  (521)
1 site		 R T G C G C A Y Y A C G C G T R
PaqCI  (585)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
AfeI  (758)
1 site		 A G C G C T T C G C G A
BstBI  (1770)
1 site		 T T C G A A A A G C T T
StuI  (1868)
1 site		 A G G C C T T C C G G A
PshAI  (2467)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PaeR7I  (2654)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2654)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2654)
1 site		 C T C G A G G A G C T C
BspEI  (3212)
1 site		 T C C G G A A G G C C T
NcoI  (3517)
1 site		 C C A T G G G G T A C C
BbvCI  (3941)
1 site		 C C T C A G C G G A G T C G
SrfI  (4087)
1 site		 G C C C G G G C C G G G C C C G
BstXI  (4204)
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.
AvrII  (4476)
1 site		 C C T A G G G G A T C C
PacI  (4523)
1 site		 T T A A T T A A A A T T A A T T
SpeI  (4535)
1 site		 A C T A G T T G A T C A
MfeI  (4839)
1 site		 C A A T T G G T T A A C
BstEII  (5111)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
AhdI  (5241)
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.
SbfI  (5266)
1 site		 C C T G C A G G G G A C G T C C
HindIII  (5274)
1 site		 A A G C T T T T C G A A
PvuI  (5400)
1 site		 C G A T C G G C T A G C
pVS1 RepA
7884 .. 8957  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
pVS1 RepA
7884 .. 8957  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
KanR
10,562 .. 11,356  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
10,562 .. 11,356  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
OCS terminator
4551 .. 5258  =  708 bp
octopine synthase terminator
OCS terminator
4551 .. 5258  =  708 bp
octopine synthase terminator
CmR
3003 .. 3662  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3003 .. 3662  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
pVS1 StaA
6826 .. 7455  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
pVS1 StaA
6826 .. 7455  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
ori
9887 .. 10,475  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9887 .. 10,475  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BlpR
373 .. 924  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
373 .. 924  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
MAS promoter
930 .. 1310  =  381 bp
mannopine synthase promoter (Velten et al., 1984)
MAS promoter
930 .. 1310  =  381 bp
mannopine synthase promoter (Velten et al., 1984)
CaMV 35S promoter
2306 .. 2651  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
2306 .. 2651  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
ccdB
4004 .. 4309  =  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
4004 .. 4309  =  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.
MAS terminator
111 .. 363  =  253 bp
mannopine synthase terminator
MAS terminator
111 .. 363  =  253 bp
mannopine synthase terminator
pVS1 oriV
9023 .. 9217  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 oriV
9023 .. 9217  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
bom
9561 .. 9701  =  141 bp
basis of mobility region from pBR322
bom
9561 .. 9701  =  141 bp
basis of mobility region from pBR322
attR2
4350 .. 4474  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4350 .. 4474  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2770 .. 2889  =  120 bp
recombination site for the Gateway® LR reaction
attR1
2770 .. 2889  =  120 bp
recombination site for the Gateway® LR reaction
TMV Ω
2681 .. 2735  =  55 bp
translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)
TMV Ω
2681 .. 2735  =  55 bp
translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)
lac UV5 promoter
2919 .. 2949  =  31 bp
3 segments
   Segment 1:  -35  
   2919 .. 2924  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2919 .. 2949  =  31 bp
3 segments
   Segment 2:  
   2925 .. 2942  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2919 .. 2949  =  31 bp
3 segments
   Segment 3:  -10  
   2943 .. 2949  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2919 .. 2949  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ATG
2737 .. 2739  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2737 .. 2739  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
AcV5 tag
2740 .. 2766  =  27 bp
9 amino acids  =  1.0 kDa
Product: epitope tag from the baculovirus Autographa californica GP64 envelope fusion protein
AcV5 tag
2740 .. 2766  =  27 bp
9 amino acids  =  1.0 kDa
Product: epitope tag from the baculovirus Autographa californica GP64 envelope fusion protein
LB T-DNA repeat
1 .. 25  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
1 .. 25  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
5502 .. 5526  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
5502 .. 5526  =  25 bp
right border repeat from nopaline C58 T-DNA
ORF:  448 .. 957  =  510 bp
ORF:  169 amino acids  =  18.0 kDa
ORF:  6826 .. 7455  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  275 .. 607  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  1730 .. 2134  =  405 bp
ORF:  134 amino acids  =  15.5 kDa
ORF:  4004 .. 4309  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  5138 .. 5428  =  291 bp
ORF:  96 amino acids  =  10.1 kDa
ORF:  1809 .. 2072  =  264 bp
ORF:  87 amino acids  =  10.4 kDa
ORF:  2124 .. 2552  =  429 bp
ORF:  142 amino acids  =  15.9 kDa
ORF:  3003 .. 3662  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  5841 .. 6527  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  7782 .. 8957  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  10,695 .. 10,925  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  6564 .. 6878  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  7752 .. 8324  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  1757 .. 2125  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5873 .. 6223  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  7730 .. 7987  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  10,562 .. 11,356  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  11,601 .. 321  =  501 bp
ORF:  166 amino acids  =  18.5 kDa
ORF:  373 .. 924  =  552 bp
ORF:  183 amino acids  =  20.6 kDa
ORF:  6292 .. 6564  =  273 bp
ORF:  90 amino acids  =  9.9 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

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