pSIREN-RetroQ-TetP

Linearized retroviral vector with a puromycin resistance marker for inserting an shRNA cassette that will be expressed from a Tet-responsive promoter.

Sequence Author: Clontech (TaKaRa)

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<BamHI> (6536) BglII (6181) U6 forward sequencing primer (6104 .. 6131) AfeI (5693) ApoI (5642) PshAI (5281) AscI (5147) SnaBI (4889) BtgZI (4883) CMV enhancerSspI (4259) XmnI (4054) ScaI (3935) PvuI (3825) FspI (3677) <EcoRI> (0) AgeI (120) BlpI (283) BsmI (441) BfuAI - BspMI (519) BsiWI (586) RsrII (646) SacII (744) DraIII (1101) PaeR7I - XhoI (1191) EcoRV (1198) PvuII (1241) NheI (1340) BmtI (1344) SalI (1433) AccI (1434) SfiI (2268) AvrII (2315) BspQI - SapI (2446) AflIII - PciI (2562) pSIREN-RetroQ-TetP 6536 bp
End  (6536)
0 sites
BglII  (6181)
1 site
A G A T C T T C T A G A
AfeI  (5693)
1 site
A G C G C T T C G C G A
ApoI  (5642)
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.
PshAI  (5281)
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.
AscI  (5147)
1 site
G G C G C G C C C C G C G C G G
SnaBI  (4889)
1 site
T A C G T A A T G C A T
BtgZI  (4883)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SspI  (4259)
1 site
A A T A T T T T A T A A
XmnI  (4054)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3935)
1 site
A G T A C T T C A T G A
PvuI  (3825)
1 site
C G A T C G G C T A G C
FspI  (3677)
1 site
T G C G C A A C G C G T
Start  (0)
0 sites
AgeI  (120)
1 site
A C C G G T T G G C C A
BlpI  (283)
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.
BsmI  (441)
1 site
G A A T G C