pLenti6.3 V5-TOPO (linearized)

Linearized lentiviral expression vector with a blasticidin resistance gene with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products.

Sequence Author: Thermo Fisher (Invitrogen)

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KasI (5543) PciI (4720) AhdI (3832) ScaI (3351) DraIII (2692) SfiI (2204) BlpI (1561) MscI (1169) PmlI (1097) NarI (5544) SfoI (5545) PluTI (5547) EcoNI (6072) MfeI (6091) BbvCI - Bpu10I (6326) AleI (6479) KflI (6835) HpaI (6929) BspDI - ClaI (7052) NdeI (7246) SnaBI (7352) BsmBI - Esp3I (7601) BamHI (7657) SpeI (7663) End (7691) Start (1) PstI (16) EcoRV (19) PaeR7I - PspXI - XhoI (40) PspOMI (52) ApaI (56) BstBI (63) MluI (107) PflMI (374) PfoI (406) CsiI - SexAI * (804) TspMI - XmaI (1058) SmaI (1060) BclI * (1088) pLenti6.3/V5-TOPO® 7690 bp
KasI  (5543)
1 site
G G C G C C C C G C G G
PciI  (4720)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (3832)
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.
ScaI  (3351)
1 site
A G T A C T T C A T G A
DraIII  (2692)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
SfiI  (2204)
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.
BlpI  (1561)
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.
MscI  (1169)
1 site
T G G C C A A C C G G T
PmlI  (1097)
1 site
C A C G T G G T G C A C
NarI  (5544)
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  (5545)
1 site
G G C G C C C C G C G G
PluTI  (5547)
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.
EcoNI  (6072)
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.
MfeI  (6091)
1 site
C A A T T G G T T A A C
BbvCI  (6326)
1 site
C C T C A G C G G A G T C G
Bpu10I  (6326)
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.
AleI  (6479)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (6835)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
HpaI  (6929)
1 site
G T T A A C C A A T T G
BspDI  (7052)
1 site
A T C G A T T A G C T A
ClaI  (7052)
1 site
A T C G A T T