pSB11

Superbinary intermediate vector for plant cell transformation, with a spectinomycin-resistance gene.
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HindIII (1) PmeI (6185) EagI (5799) SspI (5472) ScaI (5148) SbfI (4913) BmgBI (4778) PpuMI (4714) FspI (4453) AflIII - PciI (3338) BspQI - SapI (3222) PaeR7I - PspXI - XhoI (13) XbaI (19) BamHI (25) TspMI - XmaI (30) SmaI (32) Acc65I (34) KpnI (38) Eco53kI (42) SacI (44) EcoRI (46) LB T-DNA repeat BsiWI (568) PvuII (585) SpeI (669) StuI (706) MfeI (1150) BssHII (1368) BlpI (1781) BstEII (1928) streptothricin acetyltransferase PaqCI (2655) HpaI (2770) BstZ17I (3111) pSB11 6323 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
PmeI  (6185)
1 site
G T T T A A A C C A A A T T T G
EagI  (5799)
1 site
C G G C C G G C C G G C
SspI  (5472)
1 site
A A T A T T T T A T A A
ScaI  (5148)
1 site
A G T A C T T C A T G A
SbfI  (4913)
1 site
C C T G C A G G G G A C G T C C
BmgBI  (4778)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
PpuMI  (4714)
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.
FspI  (4453)
1 site
T G C G C A A C G C G T
AflIII  (3338)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3338)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3222)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3222)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PaeR7I  (13)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (13)
1 site
V C T C G A G B B G A G C T C V
XhoI  (13)
1 site
C T C G A G G A G C T C
XbaI  (19)
1 site
T C T A G A A G A T C T
BamHI  (25)
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.
TspMI  (30)
1 site
C C C G G G G G G C C C
XmaI  (30)
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  (32)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (34)
1 site
G G T A C C C C A T G G
KpnI  (38)
1 site
G G T A C C C C A T G G
Eco53kI  (42)
1 site
G A G C T C C T C G A G
SacI  (44)
1 site
G A G C T C C T C G A G
EcoRI  (46)
1 site
G A A T T C C T T A A G
BsiWI  (568)
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.
PvuII  (585)
1 site
C A G C T G G T C G A C
SpeI  (669)
1 site
A C T A G T T G A T C A
StuI  (706)
1 site
A G G C C T T C C G G A
MfeI  (1150)
1 site
C A A T T G G T T A A C
BssHII  (1368)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BlpI  (1781)
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.
BstEII  (1928)
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.
PaqCI  (2655)
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.
HpaI  (2770)
1 site
G T T A A C C A A T T G
BstZ17I  (3111)
1 site
G T A T A C C A T A T G
SmR
1304 .. 2092  =  789 bp
262 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
SmR
1304 .. 2092  =  789 bp
262 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
ori
3399 .. 3987  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3399 .. 3987  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
streptothricin acetyltransferase
2150 .. 2674  =  525 bp
174 amino acids  =  19.7 kDa
Product: streptothricin acetyltransferase
streptothricin acetyltransferase
2150 .. 2674  =  525 bp
174 amino acids  =  19.7 kDa
Product: streptothricin acetyltransferase
cos
4495 .. 4893  =  399 bp
lambda cos site; allows packaging into phage lambda particles
cos
4495 .. 4893  =  399 bp
lambda cos site; allows packaging into phage lambda particles
bom
3073 .. 3213  =  141 bp
basis of mobility region from pBR322
bom
3073 .. 3213  =  141 bp
basis of mobility region from pBR322
MCS
1 .. 51  =  51 bp
multiple cloning site
MCS
1 .. 51  =  51 bp
multiple cloning site
LB T-DNA repeat
104 .. 128  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
104 .. 128  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6147 .. 6171  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6147 .. 6171  =  25 bp
right border repeat from nopaline C58 T-DNA
ORF:  1003 .. 1230  =  228 bp
ORF:  75 amino acids  =  8.7 kDa
ORF:  4288 .. 4599  =  312 bp
ORF:  103 amino acids  =  11.0 kDa
ORF:  1670 .. 2053  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  1515 .. 1793  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  1803 .. 2060  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  2241 .. 2492  =  252 bp
ORF:  83 amino acids  =  9.3 kDa
ORF:  4158 .. 4469  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  731 .. 1228  =  498 bp
ORF:  165 amino acids  =  18.0 kDa
ORF:  1304 .. 2092  =  789 bp
ORF:  262 amino acids  =  29.2 kDa
ORF:  2150 .. 2674  =  525 bp
ORF:  174 amino acids  =  19.7 kDa
ORF:  4847 .. 5455  =  609 bp
ORF:  202 amino acids  =  22.2 kDa
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