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pGL4.14[luc2/Hygro]

Promoterless vector for measuring the activity of promoter and enhancer sequences with a luciferase assay.

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pGL4.14[luc2 Hygro].dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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AarI - BfuAI - BspMI (5811) RVprimer3 (5790 .. 5809) BsmBI (5638) SpeI (5625) BstZ17I (5306) SacII (5190) PvuI (5166) Bsu36I (5152) BstEII (4721) BstXI - PstI (4718) NotI (4694) AlwNI (4290) ApaLI (4188) BciVI (4077) PciI (3874) RVprimer4 (3675 .. 3694) SalI (3624) BstBI (3610) PmeI (3534) BssHII (3509) 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) MreI - SgrAI (163) BsrGI (590) BbvCI (811) KasI (1047) NarI (1048) SfoI (1049) PluTI (1051) BlpI (1054) DraIII (1246) BpmI (1480) FseI (1776) PsiI (1897) MfeI (1926) BamHI (2019) StuI (2452) AvrII (2453) XmnI (2536) BsaAI (2684) pGL4.14[luc2/Hygro] 5841 bp
AarI  (5811)
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  (5811)
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  (5811)
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  (5638)
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  (5625)
1 site
A C T A G T T G A T C A
BstZ17I  (5306)
1 site
G T A T A C C A T A T G
SacII  (5190)
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  (5166)
1 site
C G A T C G G C T A G C
Bsu36I  (5152)
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.
BstEII  (4721)
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  (4718)
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  (4718)
1 site
C T G C A G G A C G T C
NotI  (4694)
1 site
G C G G C C G C C G C C G G C G
AlwNI  (4290)
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.
ApaLI  (4188)
1 site
G T G C A C C A C G T G
BciVI  (4077)
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.
PciI  (3874)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (3624)
1 site
G T C G A C C A G C T G
BstBI  (3610)
1 site
T T C G A A A A G C T T
PmeI  (3534)
1 site
G T T T A A A C C A A A T T T G
BssHII  (3509)
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.
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
MreI  (163)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (163)
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.
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
KasI  (1047)
1 site
G G C G C C C C G C G G
NarI  (1048)
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  (1049)
1 site
G G C G C C C C G C G G
PluTI  (1051)
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.
BlpI  (1054)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
DraIII  (1246)
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.
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.
FseI  (1776)
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  (1897)
1 site
T T A T A A A A T A T T
MfeI  (1926)
1 site
C A A T T G G T T A A C
BamHI  (2019)
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  (2452)
1 site
A G G C C T T C C G G A
AvrII  (2453)
1 site
C C T A G G G G A T C C
XmnI  (2536)
1 site