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pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS

Vector encoding a single-chain anti-GCN4 peptide antibody fused to superfolder GFP, the VP64 activation domain, and the GB1 solubilization domain.

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pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Vale Lab / Addgene #60904
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NotI (9846) SpeI (9776) BstBI (9287) BstZ17I (9115) PspXI (9086) MscI (8836) BamHI (8644) BlpI (8270) AarI (7892) SmaI (7768) TspMI - XmaI (7766) PflFI - Tth111I (7759) BspDI * - ClaI * (7501) KflI (7230) BbvCI (6720) NruI * (6134) BssHII (6012) PmlI (5591) XcmI (5205) NheI (357) BmtI (361) NgoMIV (390) NaeI (392) AfeI (485) EcoNI (615) PfoI (779) PvuI (1449) AhdI (1818) SfiI (3163) PshAI (4040) PflMI (4434) HpaI (4998) pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS 10,074 bp
NotI  (9846)
1 site
G C G G C C G C C G C C G G C G
SpeI  (9776)
1 site
A C T A G T T G A T C A
BstBI  (9287)
1 site
T T C G A A A A G C T T
BstZ17I  (9115)
1 site
G T A T A C C A T A T G
PspXI  (9086)
1 site
V C T C G A G B B G A G C T C V
MscI  (8836)
1 site
T G G C C A A C C G G T
BamHI  (8644)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BlpI  (8270)
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.
AarI  (7892)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
SmaI  (7768)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (7766)
1 site
C C C G G G G G G C C C
XmaI  (7766)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
PflFI  (7759)
1 site
G A C