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pRetroX-Tet3G

Retroviral vector to be used in conjunction with a Tet-responsive vector for doxycycline-inducible expression of a gene.

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pRetroX-Tet3G.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SspI (7791) ScaI (7467) PvuI (7357) AlwNI (6510) SfiI (5800) BmtI (4876) NheI (4872) EcoRV (4730) BsgI (4639) PaeR7I - PspXI - XhoI (4589) RsrII (4275) NaeI (4261) AscI (542) PshAI (676) BstEII (1252) Bpu10I (1455) BglII (1576) SbfI (2230) NotI (2243) AgeI (2250) BsiWI (2262) PacI (2273) BamHI (2278) XcmI (2295) NruI (2570) MfeI (2807) EcoRI (3037) AarI (3384) PflMI (3498) BmgBI (3587) BstXI (3630) NgoMIV (4259) pRetroX-Tet3G 8136 bp
SspI  (7791)
1 site
A A T A T T T T A T A A
ScaI  (7467)
1 site
A G T A C T T C A T G A
PvuI  (7357)
1 site
C G A T C G G C T A G C
AlwNI  (6510)
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.
SfiI  (5800)
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.
BmtI  (4876)
1 site
G C T A G C C G A T C G
NheI  (4872)
1 site
G C T A G C C G A T C G
EcoRV  (4730)
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.
BsgI  (4639)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PaeR7I  (4589)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4589)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4589)
1 site
C T C G A G G A G C T C
RsrII  (4275)
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.
NaeI  (4261)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
AscI  (542)
1 site
G G C G C G C C C C G C G C G G
PshAI  (676)
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.
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.
Bpu10I  (1455)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
BglII  (1576)
1 site
A G A T C T T C T A G A
SbfI  (2230)
1 site
C C T G C A G G G G A C G T C C