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Plasmid Files

pGA643

Binary Ti vector for expression of foreign genes in plant cells.

 
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 PmeI (39) NotI (11,010) PasI (10,458) AgeI (9831) ApaI (8048) PspOMI (8044) BstXI (7685) StuI (7446) AvrII (7396) FspAI (6736) Bts α I (6261) SexAI * (6168) AleI (6080) Bsu36I (247) BamHI (408) PstI (573) AflII (1840) HindIII (2824) XbaI (2829) HpaI (2841) MCS BglII (2857) gene 7 terminator BstZ17I (3092) EcoRI (3620) AhdI (4009) SgrDI (4112) SfiI (5396) NdeI (5954) pGA643 11,601 bp
PmeI  (39)
1 site
G T T T A A A C C A A A T T T G
NotI  (11,010)
1 site
G C G G C C G C C G C C G G C G
PasI  (10,458)
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.
AgeI  (9831)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
ApaI  (8048)
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  (8044)
1 site
G G G C C C C C C G G G
BstXI  (7685)
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.
StuI  (7446)
1 site
A G G C C T T C C G G A
AvrII  (7396)
1 site
C C T A G G G G A T C C
FspAI  (6736)
1 site
R T G C G C A Y Y A C G C G T R
BtsαI  (6261)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
SexAI  (6168)
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.
AleI  (6080)
1 site
C A C N N N N G T G G T G N N N N C A C
Bsu36I  (247)
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.
BamHI  (408)
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.
PstI  (573)
1 site
C T G C A G G A C G T C
AflII  (1840)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
HindIII  (2824)
1 site
A A G C T T T T C G A A
XbaI  (2829)
1 site
T C T A G A A G A T C T
HpaI  (2841)
1 site
G T T A A C C A A T T G
BglII  (2857)
1 site
A G A T C T T C T A G A
BstZ17I  (3092)
1 site
G T A T A C C A T A T G
EcoRI  (3620)
1 site
G A A T T C C T T A A G
AhdI  (4009)
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.
SgrDI  (4112)
1 site
C G T C G A C G G C A G C T G C
SfiI  (5396)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
NdeI  (5954)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
TcR
7463 .. 8662  =  1200 bp
399 amino acids  =  42.2 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
7463 .. 8662  =  1200 bp
399 amino acids  =  42.2 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
trfA
5042 .. 6190  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds
to and activates oriV
trfA
5042 .. 6190  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds
to and activates oriV
KanR
365 .. 1186  =  822 bp
273 amino acids  =  29.9 kDa
   Segment 1:  nos  
   365 .. 406  =  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
365 .. 1186  =  822 bp
273 amino acids  =  29.9 kDa
   Segment 2:  
   407 .. 415  =  9 bp
   3 amino acids  =  443.6 Da
Product: fusion between an N-terminal peptide of
nopaline synthase and Tn5 aminoglycoside
phosphotransferase
confers resistance to kanamycin
KanR
365 .. 1186  =  822 bp
273 amino acids  =  29.9 kDa
   Segment 3:  nptII  
   416 .. 1186  =  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
365 .. 1186  =  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
TetR
8768 .. 9418  =  651 bp
216 amino acids  =  23.3 kDa
Product: tetracycline resistance regulatory protein
TetR
8768 .. 9418  =  651 bp
216 amino acids  =  23.3 kDa
Product: tetracycline resistance regulatory protein
oriV
10,483 .. 11,114  =  632 bp
incP origin of replication
oriV
10,483 .. 11,114  =  632 bp
incP origin of replication
traJ
4399 .. 4770  =  372 bp
123 amino acids  =  13.5 kDa
Product: oriT-recognizing protein
traJ
4399 .. 4770  =  372 bp
123 amino acids  =  13.5 kDa
Product: oriT-recognizing protein
CaMV 35S promoter
2471 .. 2815  =  345 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
2471 .. 2815  =  345 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
1811 .. 2063  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
1811 .. 2063  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS promoter
149 .. 364  =  216 bp
nopaline synthase promoter
NOS promoter
149 .. 364  =  216 bp
nopaline synthase promoter
gene 7 terminator
2875 .. 3075  =  201 bp
transcription terminator for octopine-type Ti plasmid
gene 7
gene 7 terminator
2875 .. 3075  =  201 bp
transcription terminator for octopine-type Ti plasmid
gene 7
oriT
4257 .. 4366  =  110 bp
incP origin of transfer
oriT
4257 .. 4366  =  110 bp
incP origin of transfer
MCS
2824 .. 2862  =  39 bp
multiple cloning site
MCS
2824 .. 2862  =  39 bp
multiple cloning site
RB T-DNA repeat
1 .. 25  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
1 .. 25  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
3678 .. 3702  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
3678 .. 3702  =  25 bp
left border repeat from nopaline C58 T-DNA
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