pCAMBIA3300

Agrobacterium binary vector for plant transformation, with bialophos/phosphinothricin resistance and kanamycin resistance genes.

Sequence Author: Cambia

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lac operator BstXI (8186) CaMV 35S promoter (enhanced) AfeI (7226) ApaI (7170) PspOMI (7166) PaqCI (7053) FspAI (6989) PspXI (6835) BclI * (6320) PsiI (6189) Bpu10I (6044) PpuMI (5905) BspHI (5863) BlpI (5713) SspI (5367) NsiI (5329) BstZ17I (4376) PluTI (4181) EcoRI (0) Eco53kI (8) SacI (10) TspMI - XmaI (16) SmaI (18) BamHI (21) XbaI (27) PstI - SbfI (43) HindIII (51) PvuI (177) PmeI (264) BsaBI * (1715) PasI (1722) AclI (2316) BsiWI (3276) NheI (3392) BmtI (3396) BspDI * - ClaI * (3472) EcoNI (3705) BsaI (3795) AgeI (3886) MreI - SgrAI (4174) KasI (4177) NarI (4178) SfoI (4179) pCAMBIA3300 8429 bp
BstXI  (8186)
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.
AfeI  (7226)
1 site		 A G C G C T T C G C G A
ApaI  (7170)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (7166)
1 site		 G G G C C C C C C G G G
PaqCI  (7053)
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.
FspAI  (6989)
1 site		 R T G C G C A Y Y A C G C G T R
PspXI  (6835)
1 site		 V C T C G A G B B G A G C T C V
BclI  (6320)
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  (6189)
1 site		 T T A T A A A A T A T T
Bpu10I  (6044)
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.
PpuMI  (5905)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
BspHI  (5863)
1 site		 T C A T G A A G T A C T
BlpI  (5713)
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.
SspI  (5367)
1 site		 A A T A T T T T A T A A
NsiI  (5329)
1 site		 A T G C A T T A C G T A
BstZ17I  (4376)
1 site		 G T A T A C C A T A T G
PluTI  (4181)
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.
EcoRI  (0)
1 site		 G A A T T C C T T A A G
Eco53kI  (8)
1 site		 G A G C T C C T C G A G
SacI  (10)
1 site		 G A G C T C C T C G A G
TspMI  (16)
1 site		 C C C G G G G G G C C C
XmaI  (16)
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.
SmaI  (18)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BamHI  (21)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
XbaI  (27)
1 site		 T C T A G A A G A T C T
PstI  (43)
1 site		 C T G C A G G A C G T C
SbfI  (43)
1 site		 C C T G C A G G G G A C G T C C
HindIII  (51)
1 site		 A A G C T T T T C G A A
PvuI  (177)
1 site		 C G A T C G G C T A G C
PmeI  (264)
1 site		 G T T T A A A C C A A A T T T G
BsaBI  (1715)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
PasI  (1722)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
AclI  (2316)
1 site		 A A C G T T T T G C A A
BsiWI  (3276)
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.
NheI  (3392)
1 site		 G C T A G C C G A T C G
BmtI  (3396)
1 site		 G C T A G C C G A T C G
BspDI  (3472)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3472)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoNI  (3705)
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.
BsaI  (3795)
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.
AgeI  (3886)
1 site		 A C C G G T T G G C C A
MreI  (4174)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (4174)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
KasI  (4177)
1 site		 G G C G C C C C G C G G
NarI  (4178)
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.
SfoI  (4179)
1 site		 G G C G C C C C G C G G
pVS1 RepA
2661 .. 3734  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
pVS1 RepA
2661 .. 3734  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
KanR
5339 .. 6133  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
5339 .. 6133  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
7437 .. 8114  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
7437 .. 8114  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
pVS1 StaA
1603 .. 2232  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
pVS1 StaA
1603 .. 2232  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
ori
4664 .. 5252  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4664 .. 5252  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BlpR
6841 .. 7392  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
6841 .. 7392  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
lacZα
8415 .. 219  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8415 .. 219  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
3800 .. 3994  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 oriV
3800 .. 3994  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6660 .. 6834  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6660 .. 6834  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
4338 .. 4478  =  141 bp
basis of mobility region from pBR322
bom
4338 .. 4478  =  141 bp
basis of mobility region from pBR322
lac promoter
8341 .. 8371  =  31 bp
3 segments
   Segment 1:  -35  
   8341 .. 8346  =  6 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
3 segments
   Segment 2:  
   8347 .. 8364  =  18 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
3 segments
   Segment 3:  -10  
   8365 .. 8371  =  7 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
279 .. 303  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
279 .. 303  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6558 .. 6582  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6558 .. 6582  =  25 bp
left border repeat from nopaline C58 T-DNA
lac operator
8379 .. 8395  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
8379 .. 8395  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
MCS
8429 .. 56  =  57 bp
pUC18/19 multiple cloning site
MCS
8429 .. 56  =  57 bp
pUC18/19 multiple cloning site
M13 fwd
60 .. 76  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
60 .. 76  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8403 .. 8419  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8403 .. 8419  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1603 .. 2232  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  6826 .. 7434  =  609 bp
ORF:  202 amino acids  =  21.8 kDa
ORF:  618 .. 1304  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  2559 .. 3734  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  5472 .. 5702  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  8415 .. 219  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  1341 .. 1655  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2529 .. 3101  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  650 .. 1000  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  2507 .. 2764  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  5339 .. 6133  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  7541 .. 7858  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  7868 .. 8326  =  459 bp
ORF:  152 amino acids  =  16.9 kDa
ORF:  1069 .. 1341  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  6841 .. 7515  =  675 bp
ORF:  224 amino acids  =  25.4 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