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pCLuc Mini-TK 2

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

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pCLuc Mini-TK 2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  New England Biolabs
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AatII (6130) SgrDI (6129) ZraI (6128) SspI (6012) ScaI (5688) TatI (5686) PvuI (5578) BglI (5328) BsaI (5269) AhdI (5208) BstZ17I (3936) BstBI (3646) RsrII (3480) BssHII (3361) MscI (3046) EcoRI (20) EcoRV (32) PaeR7I - PspXI - XhoI (36) HindIII (45) Acc65I (51) KpnI (55) Eco53kI (59) SacI (61) BamHI (63) MluI (85) BsgI (276) AarI (298) XcmI (543) BstEII (654) PshAI (887) PmlI (1041) BmgBI (1064) EcoO109I - KflI - PpuMI (1153) Bsu36I (1705) NotI (1808) XbaI (1864) DraIII (2230) StuI (2753) AvrII (2754) TspMI - XmaI (2775) SmaI (2777) KasI (2963) NarI (2964) SfoI (2965) PluTI (2967) pCLuc Mini-TK 2 6131 bp
AatII  (6130)
1 site
G A C G T C C T G C A G
SgrDI  (6129)
1 site
C G T C G A C G G C A G C T G C
ZraI  (6128)
1 site
G A C G T C C T G C A G
SspI  (6012)
1 site
A A T A T T T T A T A A
ScaI  (5688)
1 site
A G T A C T T C A T G A
TatI  (5686)
1 site
W G T A C W W C A T G W
PvuI  (5578)
1 site
C G A T C G G C T A G C
BglI  (5328)
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  (5269)
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.
AhdI  (5208)
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.
BstZ17I  (3936)
1 site
G T A T A C C A T A T G
BstBI  (3646)
1 site
T T C G A A A A G C T T
RsrII  (3480)
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.
BssHII  (3361)
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.
MscI  (3046)
1 site
T G G C C A A C C G G T
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
BsgI  (276)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AarI  (298)
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.
XcmI  (543)
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.
BstEII  (654)
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.
PshAI  (887)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PmlI  (1041)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BmgBI  (1064)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
EcoO109I  (1153)
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.
KflI  (1153)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1153)
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.
Bsu36I  (1705)
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.
NotI  (1808)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1864)
1 site
T C T A G A A G A T C T
DraIII  (2230)
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.
StuI  (2753)
1 site
A G G C C T T C C G G A
AvrII  (2754)
1 site
C C