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Plasmid Files

pSP-luc+NF

Enhanced firefly luciferase vector for creating N-terminal fusions and for in vitro transcription.

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pSP-luc+NF.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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 Acc65I (11) NdeI (4020) AatII (3771) ZraI (3769) SspI (3653) XmnI (3448) PvuI (3219) FspI (3071) NmeAIII (2997) BglI (2969) AlwNI (2372) KpnI (15) NheI (17) BmtI (21) BglII (24) AvrII (30) HindIII (36) NcoI (43) BstEII (47) KasI (80) NarI (81) SfoI (82) PluTI (84) BstBI (217) PsiI (373) BsrGI (538) BclI * (628) SphI (711) XcmI (783) BanII (1072) PpuMI (1227) AarI (1437) SgrAI (1476) XbaI (1702) EcoRI (1708) EcoRV (1716) BspDI * - ClaI * (1721) PaeR7I - XhoI (1726) HpaI (1777) PciI (1956) pSP-luc+NF 4103 bp
Acc65I  (11)
1 site
G G T A C C C C A T G G
NdeI  (4020)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
AatII  (3771)
1 site
G A C G T C C T G C A G
ZraI  (3769)
1 site
G A C G T C C T G C A G
SspI  (3653)
1 site
A A T A T T T T A T A A
XmnI  (3448)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3219)
1 site
C G A T C G G C T A G C
FspI  (3071)
1 site
T G C G C A A C G C G T
NmeAIII  (2997)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (2969)
1 site
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.
AlwNI  (2372)
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.
KpnI  (15)
1 site
G G T A C C C C A T G G
NheI  (17)
1 site
G C T A G C C G A T C G
BmtI  (21)
1 site
G C T A G C C G A T C G
BglII  (24)
1 site
A G A T C T T C T A G A
AvrII  (30)
1 site
C C T A G G G G A T C C
HindIII  (36)
1 site
A A G C T T T T C G A A
NcoI  (43)
1 site
C C A T G G G G T A C C
BstEII  (47)
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.
KasI  (80)
1 site
G G C G C C C C G C G G
NarI  (81)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (82)
1 site
G G C G C C C C G C G G
PluTI  (84)
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.
BstBI  (217)
1 site
T T C G A A A A G C T T
PsiI  (373)
1 site
T T A T A A A A T A T T
BsrGI  (538)
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.
BclI  (628)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
SphI  (711)
1 site
G C A T G C C G T A C G
XcmI  (783)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BanII  (1072)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PpuMI  (1227)
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.
AarI  (1437)
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.
SgrAI  (1476)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
XbaI  (1702)
1 site
T C T A G A A G A T C T
EcoRI  (1708)
1 site
G A A T T C C T T A A G
EcoRV  (1716)
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.
BspDI  (1721)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1721)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PaeR7I  (1726)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1726)
1 site
C T C G A G G A G C T C
HpaI  (1777)
1 site
G T T A A C C A A T T G
PciI  (1956)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
luciferase
45 .. 1700  =  1656 bp
551 amino acids  =  60.7 kDa
Product: firefly luciferase
enhanced luc+NF version of the luciferase gene
luciferase
45 .. 1700  =  1656 bp
551 amino acids  =  60.7 kDa
Product: firefly luciferase
enhanced luc+NF version of the luciferase gene
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2776 .. 3567  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3568 .. 3636  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2017 .. 2605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2017 .. 2605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
3637 .. 3741  =  105 bp
AmpR promoter
3637 .. 3741  =  105 bp
downstream MCS
1702 .. 1731  =  30 bp
downstream multiple cloning site
downstream MCS
1702 .. 1731  =  30 bp
downstream multiple cloning site
T7 promoter
1741 .. 1759  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1741 .. 1759  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
4087 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
4087 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
upstream MCS
11 .. 53  =  43 bp
upstream multiple cloning site
upstream MCS
11 .. 53  =  43 bp
upstream multiple cloning site
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