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Plasmid Files

pLATE11 (linearized)

Vector for ligation independent cloning (LIC) and tightly regulated bacterial expression of an untagged protein.

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pLATE11 (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher
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AatII (4004) ZraI (4002) SspI (3886) ScaI (3562) PvuI (3452) FspI (3304) BglI (3202) AhdI (3082) AlwNI (2605) PciI (2189) BspQI - SapI (2073) NdeI (2012) BstZ17I (1962) AccI (1961) BsaAI (1943) PflFI - Tth111I (1936) PfoI (1833) AfeI (1445) Eco53kI (4315) SacI (4317) Acc65I (4319) KpnI (4323) MauBI (4330) AarI - BfuAI - BspMI (4335) SbfI (4346) AscI (4351) LIC forward sequencing primer (4360 .. 4379) XbaI (4407) SwaI (4432) End (4438) Start (0) PmeI (18) StuI (25) SphI (33) PacI (55) LIC reverse sequencing primer (62 .. 85) HindIII (123) BspDI - ClaI (130) BamHI (157) StyI (173) PflMI (228) MluI (646) BclI * (660) BstEII (827) PspOMI (853) ApaI (857) ApoI (921) EcoRV (1096) HpaI (1152) KasI (1285) NarI * (1286) SfoI (1287) PluTI (1289) BspEI * (1380) BsaBI * (1388) pLATE11 4438 bp
AatII  (4004)
1 site
G A C G T C C T G C A G
ZraI  (4002)
1 site
G A C G T C C T G C A G
SspI  (3886)
1 site
A A T A T T T T A T A A
ScaI  (3562)
1 site
A G T A C T T C A T G A
PvuI  (3452)
1 site
C G A T C G G C T A G C
FspI  (3304)
1 site
T G C G C A A C G C G T
BglI  (3202)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
AhdI  (3082)
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.
AlwNI  (2605)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PciI  (2189)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2073)
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  (2073)
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.
NdeI  (2012)
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.
BstZ17I  (1962)
1 site
G T A T A C C A T A T G
AccI  (1961)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BsaAI  (1943)
1 site
Y A C G T R R T G C A Y
PflFI  (1936)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1936)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PfoI  (1833)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AfeI  (1445)
1 site
A G C G C T T C G C G A
Eco53kI  (4315)
1 site
G A G C T C C T C G A G
SacI  (4317)
1 site
G A G C T C C T C G A G
Acc65I  (4319)
1 site
G G T A C C C C A T G G
KpnI  (4323)
1 site
G G T A C C C C A T G G
MauBI  (4330)
1 site
C G C G C G C G G C G C G C G C
AarI  (4335)
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 AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (4335)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (4335)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SbfI  (4346)
1 site
C C T G C A G G G G A C G T C C
AscI  (4351)
1 site
G G C G C G C C C C G C G C G G
XbaI  (4407)
1 site
T C T A G A A G A T C T
SwaI  (4432)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
End  (4438)
0 sites
Start  (0)
0 sites
PmeI  (18)
1 site
G T T T A A A C C A A A T T T G
StuI  (25)
1 site
A G G C C T T C C G G A
SphI  (33)
1 site
G C A T G C C G T A C G
PacI  (55)
1 site
T T A A T T A A A A T T A A T T
HindIII  (123)
1 site
A A G C T T T T C G A A
BspDI  (130)
1 site
A T C G A T T A G C T A
ClaI  (130)
1 site
A T C G A T T A G C T A
BamHI  (157)
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.
StyI  (173)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PflMI  (228)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
MluI  (646)
1 site
A C G C G T T G C G C A
BclI  (660)
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.
BstEII  (827)
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.
PspOMI  (853)
1 site
G G G C C C C C C G G G
ApaI  (857)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
ApoI  (921)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRV  (1096)
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.
HpaI  (1152)
1 site
G T T A A C C A A T T G
KasI  (1285)
1 site
G G C G C C C C G C G G
NarI  (1286)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1287)
1 site
G G C G C C C C G C G G
PluTI  (1289)
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.
BspEI  (1380)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
BsaBI  (1388)
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.
LIC forward sequencing primer
20-mer  /  40% GC
1 binding site
4360 .. 4379  =  20 annealed bases
Tm  =  48°C
LIC reverse sequencing primer
24-mer  /  46% GC
1 binding site
62 .. 85  =  24 annealed bases
Tm  =  58°C
lacI
296 .. 1378  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
296 .. 1378  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3009 .. 3800  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3801 .. 3869  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2250 .. 2838  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2250 .. 2838  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
1631 .. 1822  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1631 .. 1822  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
3870 .. 3974  =  105 bp
AmpR promoter
3870 .. 3974  =  105 bp
rrnB T1 terminator
4075 .. 4156  =  82 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4075 .. 4156  =  82 bp
transcription terminator T1 from the E. coli rrnB gene
lacI promoter
218 .. 295  =  78 bp
lacI promoter
218 .. 295  =  78 bp
T7 terminator
162 .. 209  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
162 .. 209  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tet promoter
119 .. 147  =  29 bp
   Segment 3:  -10  
   119 .. 124  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
   Segment 2:  
   125 .. 141  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
   Segment 1:  -35  
   142 .. 147  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
rrnB T2 terminator
4248 .. 4275  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4248 .. 4275  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
4379 .. 4403  =  25 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
4379 .. 4403  =  25 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).
T7 promoter
4360 .. 4378  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4360 .. 4378  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
4293 .. 4309  =  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
4293 .. 4309  =  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).
RBS
4440 .. 4446  =  7 bp
ribosome binding site
RBS
4440 .. 4446  =  7 bp
ribosome binding site
ORF:  229 .. 546  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1135 .. 1527  =  393 bp
ORF:  130 amino acids  =  14.0 kDa
ORF:  3139 .. 3405  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  419 .. 1378  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1598 .. 1822  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  1243 .. 1599  =  357 bp
ORF:  118 amino acids  =  13.4 kDa
ORF:  3009 .. 3869  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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