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pNL3.3[secNluc/minP]

Vector with a minimal promoter for measuring the activity of transcriptional response elements using secreted NanoLuc® luciferase.

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pNL3.3[secNluc minP].dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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AarI - BfuAI - BspMI (3205) RVprimer3 (3184 .. 3203) BsmBI (3032) SpeI (3019) BsaHI (2727) BstZ17I (2700) SacII (2584) BtgI (2581) PvuI (2560) Bsu36I (2546) MscI (2424) AhdI (2190) BstEII (2115) BstXI - PstI (2112) AleI (2110) NotI (2088) BspHI (1988) AlwNI (1684) BaeGI - Bme1580I (1586) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) BanII - SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) EcoRV (41) BglII (46) BglI - SfiI (59) HindIII (65) PasI (185) PflFI - Tth111I (288) EcoNI (297) BtgZI (563) PpuMI - SanDI (616) XbaI (750) FseI (769) AanI - AanI - PsiI (890) HpaI (910) MfeI (919) BamHI (1012) SalI (1018) PshAI (1083) RVprimer4 (1069 .. 1088) AfeI (1144) BspQI - SapI (1152) AflIII - PciI (1268) NspI (1272) BciVI (1471) ApaLI (1582) pNL3.3[secNluc/minP] 3235 bp
AarI  (3205)
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.
BfuAI  (3205)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3205)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BsmBI  (3032)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
SpeI  (3019)
1 site
A C T A G T T G A T C A
BsaHI  (2727)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
BstZ17I  (2700)
1 site
G T A T A C C A T A T G
SacII  (2584)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BtgI  (2581)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
PvuI  (2560)
1 site
C G A T C G G C T A G C
Bsu36I  (2546)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
MscI  (2424)
1 site
T G G C C A A C C G G T
AhdI  (2190)
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.
BstEII  (2115)
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.
BstXI  (2112)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
PstI  (2112)
1 site
C T G C A G G A C G T C
AleI  (2110)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (2088)
1 site
G C G G C C G C C G C C G G C G
BspHI  (1988)
1 site
T C A T G A A G T A C T
AlwNI  (1684)
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.
BaeGI  (1586)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (1586)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
BglI  (8)
2 sites
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SfiI  (8)
2 sites
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.
Acc65I  (14)
1 site
G G T A C C C C A T G G
KpnI  (18)
1 site
G G T A C C C C A T G G
Eco53kI  (23)
1 site
G A G C T C C T C G A G
BanII  (25)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (25)
1 site
G A G C T C C T C G A G
NheI  (27)
1 site
G C T A G C C G A T C G
BmtI  (31)
1 site
G C T A G C C G A T C G
AbsI  (33)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (33)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (33)
1 site
V C T C G A G B B G A G C T C V
XhoI  (33)
1 site
C T C G A G G A G C T C
EcoRV  (41)
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.
BglII  (46)
1 site
A G A T C T T C T A G A
BglI  (59)
2 sites
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SfiI  (59)
2 sites
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.
HindIII  (65)
1 site
A A G C T T T T C G A A
PasI  (185)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflFI  (288)
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  (288)
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.
EcoNI  (297)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BtgZI  (563)
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.
PpuMI  (616)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
SanDI  (616)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI sites may not be compatible.
XbaI  (750)
1 site
T C T A G A A G A T C T
FseI  (769)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
AanI  (890)
1 site
T T A T A A A A T A T T
AanI  (890)
1 site
T T A T A A A A T A T T
PsiI  (890)
1 site
T T A T A A A A T A T T
HpaI  (910)
1 site
G T T A A C C A A T T G
MfeI  (919)
1 site
C A A T T G G T T A A C
BamHI  (1012)
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.
SalI  (1018)
1 site
G T C G A C C A G C T G
PshAI  (1083)
1 site
G A C N N N N G T C C T