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pGL4.22[luc2CP/Puro]

Promoterless vector encoding highly destabilized luciferase for measuring the activity of promoter and enhancer sequences.

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pGL4.22[luc2CP Puro].dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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AarI - BfuAI - BspMI (5553) RVprimer3 (5532 .. 5551) BsmBI (5380) SpeI (5367) BstZ17I (5048) SacII (4932) PvuI (4908) Bsu36I (4894) AhdI (4538) BstEII (4463) BstXI (4460) NotI (4436) ApaLI (3930) BciVI (3819) AflIII - PciI (3616) RVprimer4 (3417 .. 3436) SalI (3366) BstBI (3352) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) EcoRV (41) BglII (46) BglI - SfiI (59) HindIII (65) PspOMI (126) ApaI (130) MreI (163) BsrGI (590) BbvCI (811) BpmI (1480) EcoRI (1749) XmnI (1768) AflII (1779) FseI (1955) PsiI (2076) MfeI (2105) BamHI (2198) StuI (2631) NruI (2715) pGL4.22[luc2CP/Puro] 5583 bp
AarI  (5553)
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  (5553)
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  (5553)
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  (5380)
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  (5367)
1 site
A C T A G T T G A T C A
BstZ17I  (5048)
1 site
G T A T A C C A T A T G
SacII  (4932)
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.
PvuI  (4908)
1 site
C G A T C G G C T A G C
Bsu36I  (4894)
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.
AhdI  (4538)
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  (4463)
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  (4460)
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.
NotI  (4436)
1 site
G C G G C C G C C G C C G G C G
ApaLI  (3930)
1 site
G T G C A C C A C G T G
BciVI  (3819)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
AflIII  (3616)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3616)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (3366)
1 site
G T C G A C C A G C T G
BstBI  (3352)
1 site
T T C G A A A A G C T T
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
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
PspOMI  (126)
1 site
G G G C C C C C C G G G
ApaI  (130)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
MreI  (163)
1 site
C G C C G G C G G C G G C C G C
BsrGI  (590)
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.
BbvCI  (811)
1 site
C C T C A G C G G A G T C G
BpmI  (1480)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
EcoRI  (1749)
1 site
G A A T T C C T T A A G
XmnI  (1768)
1 site
G A A N N N N T T C C T T N N N N A A G
AflII  (1779)
1 site
C T T A A G G A A T T C
FseI  (1955)
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.
PsiI  (2076)
1 site
T T A T A A A A T A T T
MfeI  (2105)
1 site
C A A T T G G T T A A C
BamHI  (2198)
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.
StuI  (2631)
1 site
A G G C C T T C C G G A
NruI  (2715)
1 site
T C G C G A A G C G C T
RVprimer3
20-mer  /  50% GC
1 binding site
5532 .. 5551  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
3417 .. 3436  =  20 annealed bases
Tm  =  61°C
luciferase
100 .. 1749  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
100 .. 1749  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4465 .. 5256  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5257 .. 5325  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
2678 .. 3277  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2678 .. 3277  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
3677 .. 4265  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3677 .. 4265  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
2290 .. 2647  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2290 .. 2647  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
1975 .. 2096  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1975 .. 2096  =  122 bp
SV40 polyadenylation signal
hPEST
1807 .. 1926  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
hPEST
1807 .. 1926  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
pause site
5492 .. 5583  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5492 .. 5583  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
MCS
1 .. 70  =  70 bp
multiple cloning site
MCS
1 .. 70  =  70 bp
multiple cloning site
poly(A) signal
3302 .. 3350  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3302 .. 3350  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5430 .. 5478  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5430 .. 5478  =  49 bp
synthetic polyadenylation signal
hCL1
1756 .. 1803  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring ubiquitin-dependent degradation
human codon-optimized
hCL1
1756 .. 1803  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring ubiquitin-dependent degradation
human codon-optimized
ORF:  100 .. 1929  =  1830 bp
ORF:  609 amino acids  =  67.0 kDa
ORF:  2678 .. 3277  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  4595 .. 4861  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  43 .. 327  =  285 bp
ORF:  94 amino acids  =  10.0 kDa
ORF:  4465 .. 5325  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1599 .. 1883  =  285 bp
ORF:  94 amino acids  =  10.4 kDa
ORF:  2907 .. 3185  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  2633 .. 3295  =  663 bp
ORF:  220 amino acids  =  21.9 kDa
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