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Plasmid Files

pGLuc Mini-TK 2

Mammalian vector with a minimal promoter for measuring promoter or enhancer activity using secreted Gaussia luciferase.

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pGLuc Mini-TK 2 Sequence and MappGLuc Mini-TK 2.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 AatII (5027) SgrDI (5026) ZraI (5025) SspI (4909) ScaI (4585) TatI (4583) BglI (4225) BsaI (4166) PciI (3212) BstZ17I (2833) BsmI (2781) PfoI (2636) BstBI (2543) BglII (12) EcoRI (20) EcoRV (32) PaeR7I - PspXI - XhoI (36) HindIII (45) Acc65I (51) KpnI (55) Eco53kI (59) SacI (61) BamHI (63) MluI (85) Mini-TK promoter BbsI (222) NruI (251) SacII (277) NotI (705) XbaI (761) DraIII (1127) SexAI * (1418) BseRI (1647) StuI (1650) AvrII (1651) TspMI - XmaI (1672) SmaI (1674) BclI * (1702) KasI (1860) NarI (1861) SfoI (1862) PluTI (1864) PflFI - Tth111I (1979) BssHII (2258) RsrII (2377) pGLuc Mini-TK 2 5028 bp
AatII  (5027)
1 site
G A C G T C C T G C A G
SgrDI  (5026)
1 site
C G T C G A C G G C A G C T G C
ZraI  (5025)
1 site
G A C G T C C T G C A G
SspI  (4909)
1 site
A A T A T T T T A T A A
ScaI  (4585)
1 site
A G T A C T T C A T G A
TatI  (4583)
1 site
W G T A C W W C A T G W
BglI  (4225)
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.
BsaI  (4166)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
PciI  (3212)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (2833)
1 site
G T A T A C C A T A T G
BsmI  (2781)
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  (2636)
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.
BstBI  (2543)
1 site
T T C G A A A A G C T T
BglII  (12)
1 site
A G A T C T T C T A G A
EcoRI  (20)
1 site
G A A T T C C T T A A G
EcoRV  (32)
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.
PaeR7I  (36)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (36)
1 site
V C T C G A G B B G A G C T C V
XhoI  (36)
1 site
C T C G A G G A G C T C
HindIII  (45)
1 site
A A G C T T T T C G A A
Acc65I  (51)
1 site
G G T A C C C C A T G G
KpnI  (55)
1 site
G G T A C C C C A T G G
Eco53kI  (59)
1 site
G A G C T C C T C G A G
SacI  (61)
1 site
G A G C T C C T C G A G
BamHI  (63)
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.
MluI  (85)
1 site
A C G C G T T G C G C A
BbsI  (222)
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.
NruI  (251)
1 site
T C G C G A A G C G C T
SacII  (277)
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.
NotI  (705)
1 site
G C G G C C G C C G C C G G C G
XbaI  (761)
1 site
T C T A G A A G A T C T
DraIII  (1127)
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.
SexAI  (1418)
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  (1647)
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  (1650)
1 site
A G G C C T T C C G G A
AvrII  (1651)
1 site
C C T A G G G G A T C C
TspMI  (1672)
1 site
C C C G G G G G G C C C
XmaI  (1672)
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.
SmaI  (1674)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BclI  (1702)
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.
KasI  (1860)
1 site
G G C G C C C C G C G G
NarI  (1861)
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  (1862)
1 site
G G C G C C C C G C G G
PluTI  (1864)
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.
PflFI  (1979)
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  (1979)
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.
BssHII  (2258)
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.
RsrII  (2377)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
AmpR
4032 .. 4892  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4032 .. 4823  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4032 .. 4892  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4824 .. 4892  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4032 .. 4892  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1733 .. 2527  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
1733 .. 2527  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
3273 .. 3861  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3273 .. 3861  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hGLuc
146 .. 703  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
hGLuc
146 .. 703  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
f1 ori
894 .. 1322  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
894 .. 1322  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1336 .. 1666  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1336 .. 1666  =  331 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2701 .. 2822  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2701 .. 2822  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4893 .. 4997  =  105 bp
AmpR promoter
4893 .. 4997  =  105 bp
Mini-TK promoter
69 .. 131  =  63 bp
minimal herpes simplex virus (HSV) thymidine
kinase promoter
Mini-TK promoter
69 .. 131  =  63 bp
minimal herpes simplex virus (HSV) thymidine
kinase promoter
poly(A) signal
712 .. 760  =  49 bp
synthetic polyadenylation signal
poly(A) signal
712 .. 760  =  49 bp
synthetic polyadenylation signal
SV40 ori
1517 .. 1652  =  136 bp
SV40 origin of replication
SV40 ori
1517 .. 1652  =  136 bp
SV40 origin of replication
Kozak sequence
140 .. 149  =  10 bp
Kozak sequence
140 .. 149  =  10 bp
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