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pLATE52 (linearized)

Vector for ligation independent cloning (LIC) and tightly regulated bacterial expression of a protein with a WELQut-cleavable N-terminal 6xHis tag.

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pLATE52 (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) NheI (4491) BmtI (4495) End (4526) 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) pLATE52 4526 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