pcDNA3.1 NT-GFP

Vector for expressing N-terminally GFP-tagged proteins in mammalian cells.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (6158) SspI (6041) ScaI (5717) PvuI (5607) AhdI (5237) PciI (4344) BsmI (3913) PfoI (3768) BssHII (3390) PflFI - Tth111I (3111) PluTI (2996) BglII (12) Bpu10I (180) NruI (208) SnaBI (590) HindIII (1028) BsrGI (1183) AgeI (1633) NheI (1639) BmtI (1643) Acc65I (1645) KpnI (1649) EcoRI (1680) EcoRV (1692) NotI (1707) XbaI (1719) EcoO109I - PspOMI (1725) ApaI (1729) PmeI (1734) BbsI (1945) DraIII (2259) CsiI - SexAI * (2550) BseRI (2779) StuI (2782) AvrII (2783) TspMI - XmaI (2804) SmaI (2806) BsaBI * (2852) KasI (2992) NarI (2993) SfoI (2994) pcDNA™3.1/NT-GFP 6160 bp
SgrDI  (6158)
1 site
C G T C G A C G G C A G C T G C
SspI  (6041)
1 site
A A T A T T T T A T A A
ScaI  (5717)
1 site
A G T A C T T C A T G A
PvuI  (5607)
1 site
C G A T C G G C T A G C
AhdI  (5237)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PciI  (4344)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BsmI  (3913)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
PfoI  (3768)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BssHII  (3390)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
PflFI  (3111)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3111)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PluTI  (2996)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BglII  (12)
1 site
A G A T C T T C T A G A
Bpu10I  (180)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
NruI  (208)
1 site
T C G C G A A G C G C T
SnaBI  (590)
1 site
T A C G T A A T G C A T
HindIII  (1028)
1 site
A A G C T T T T C G A A
BsrGI  (1183)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AgeI  (1633)
1 site
A C C G G T T G G C C A
NheI  (1639)
1 site
G C T A G C C G A T C G
BmtI  (1643)
1 site
G C T A G C C G A T C G
Acc65I  (1645)
1 site
G G T A C C C C A T G G
KpnI  (1649)
1 site
G G T A C C C C A T G G
EcoRI  (1680)
1 site
G A A T T C C T T A A G
EcoRV  (1692)
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.
NotI  (1707)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1719)
1 site
T C T A G A A G A T C T
EcoO109I  (1725)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (1725)
1 site
G G G C C C C C C G G G
ApaI  (1729)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmeI  (1734)
1 site
G T T T A A A C C A A A T T T G
BbsI  (1945)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
DraIII  (2259)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
CsiI  (2550)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2550)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BseRI  (2779)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
StuI  (2782)
1 site
A G G C C T T C C G G A
AvrII  (2783)
1 site
C C T A G G G G A T C C
TspMI  (2804)
1 site
C C C G G G G G G C C C
XmaI  (2804)
1 site
C C C G G G