pRetroX-Tight-Hyg

Retroviral vector for expressing a gene with the Tet-On® Advanced or Tet-Off® Advanced system.

Sequence Author: Clontech (TaKaRa)

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SspI (7074) XmnI (6869) FspI (6492) PciI (5377) BspQI - SapI (5261) AvrII (5130) SfiI (5083) PvuII (4056) EcoRV (4013) BspDI - ClaI (3828) AleI (3741) CMV enhancer SnaBI (284) AscI (542) AfeI (1088) BstEII (1252) BglII (1576) sequencing primer (1578 .. 1601) PaeR7I - PspXI - XhoI (1600) BamHI (1925) NotI (1932) NgoMIV (1939) NaeI (1941) XbaI * (1945) NruI * (1953) MluI (1957) EcoRI (1963) AgeI (2083) BsmI (2404) AsiSI (2854) RsrII (2898) BbvCI (3555) pRetroX-Tight-Hyg 7419 bp
SspI  (7074)
1 site
A A T A T T T T A T A A
XmnI  (6869)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (6492)
1 site
T G C G C A A C G C G T
PciI  (5377)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5261)
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  (5261)
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.
AvrII  (5130)
1 site
C C T A G G G G A T C C
SfiI  (5083)
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.
PvuII  (4056)
1 site
C A G C T G G T C G A C
EcoRV  (4013)
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.
BspDI  (3828)
1 site
A T C G A T T A G C T A
ClaI  (3828)
1 site
A T C G A T T A G C T A
AleI  (3741)
1 site
C A C N N N N G T G G T G N N N N C A C
SnaBI  (284)
1 site
T A C G T A A T G C A T
AscI  (542)
1 site
G G C G C G C C C C G C G C G G
AfeI  (1088)
1 site
A G C G C T T C G C G A
BstEII  (1252)
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.
BglII  (1576)
1 site
A G A T C T T C T A G A
PaeR7I  (1600)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1600)
1 site
V C T C G A G B B