pRetroX-TetOne-Puro

All-in-one retroviral vector with a puromycin resistance marker, for strong inducible expression of genes using the Tet-On® system.

Sequence Author: Clontech (TaKaRa)

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BspQI - SapI (9222) PciI (9105) AlwNI (8696) ScaI (7736) SspI (7412) AleI (6074) EcoRV (5689) PflMI (5512) RsrII (4770) SnaBI (519) AfeI (717) Eco53kI (733) SacI (735) PshAI (1170) SrfI (1394) AflII (1416) Bsu36I (1428) Bpu10I (1700) PpuMI (2077) BamHI (2607) NgoMIV (2613) NaeI (2615) SgrAI (2619) MCS BstZ17I (2627) EcoRI (2633) HindIII (2752) PaeR7I - PspXI - XhoI (3004) hPGK promoter AscI (3368) BspEI (3473) NruI (3832) BfuAI - BspMI (4109) SfiI (4583) SalI (4762) pRetroX-TetOne™-Puro 9463 bp
BspQI  (9222)
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  (9222)
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.
PciI  (9105)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (8696)
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.
ScaI  (7736)
1 site
A G T A C T T C A T G A
SspI  (7412)
1 site
A A T A T T T T A T A A
AleI  (6074)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoRV  (5689)
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.
PflMI  (5512)
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.
RsrII  (4770)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
SnaBI  (519)
1 site
T A C G T A A T G C A T
AfeI  (717)
1 site
A G C G C T T C G C G A
Eco53kI  (733)
1 site
G A G C T C C T C G A G
SacI  (735)
1 site
G A G C T C C T C G A G
PshAI  (1170)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
SrfI  (1394)
1 site
G C C C G G G C C G G G C C C G
AflII  (1416)
1 site
C T