pCBG99-Basic

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

Sequence Author: Promega

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HomePlasmidsLuciferase VectorspCBG99-Basic
BsgI (4784) BfuAI - BspMI - PaqCI (4756) RVprimer3 (4735 .. 4754) poly(A) signal NotI (4626) BsaAI (4277) XmnI (3727) AseI (3300) NmeAIII (3276) AhdI (3128) AlwNI (2651) PspFI (2543) BseYI (2539) Acc65I (1) KpnI (5) Eco53kI (9) SacI (11) NheI (21) BmtI (25) TspMI - XmaI (26) SmaI - SrfI (28) PaeR7I - XhoI (32) BglII (36) HindIII (53) BtgI - NcoI (86) PflMI (138) BstXI (292) ZraI (312) AatII (314) PfoI * (491) PspOMI (1224) ApaI (1228) EcoO109I (1258) BseRI (1456) NsiI (1468) XbaI (1717) FseI (1736) HpaI (1877) BsaBI * (1978) BamHI (1979) SalI (1985) PshAI (2050) RVprimer4 (2036 .. 2055) BspQI - SapI (2119) PciI (2235) NspI (2239) pCBG99-Basic 4793 bp
BsgI  (4784)
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).
BfuAI  (4756)
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  (4756)
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.
PaqCI  (4756)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
NotI  (4626)
1 site		 G C G G C C G C C G C C G G C G
BsaAI  (4277)
1 site		 Y A C G T R R T G C A Y
XmnI  (3727)
1 site		 G A A N N N N T T C C T T N N N N A A G
AseI  (3300)
1 site		 A T T A A T T A A T T A
NmeAIII  (3276)
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).
AhdI  (3128)
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.
AlwNI  (2651)
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.
PspFI  (2543)
1 site		 C C C A G C G G G T C G
BseYI  (2539)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
Acc65I  (1)
1 site		 G G T A C C C C A T G G
KpnI  (5)
1 site		 G G T A C C C C A T G G
Eco53kI  (9)
1 site		 G A G C T C C T C G A G
SacI  (11)
1 site		 G A G C T C C T C G A G
NheI  (21)
1 site		 G C T A G C C G A T C G
BmtI  (25)
1 site		 G C T A G C C G A T C G
TspMI  (26)
1 site		 C C C G G G G G G C C C
XmaI  (26)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (28)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SrfI  (28)
1 site		 G C C C G G G C C G G G C C C G
PaeR7I  (32)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (32)
1 site		 C T C G A G G A G C T C
BglII  (36)
1 site		 A G A T C T T C T A G A
HindIII  (53)
1 site		 A A G C T T T T C G A A
BtgI  (86)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
NcoI  (86)
1 site		 C C A T G G G G T A C C
PflMI  (138)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
BstXI  (292)
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.
ZraI  (312)
1 site		 G A C G T C C T G C A G
AatII  (314)
1 site		 G A C G T C C T G C A G
PfoI  (491)
1 site		 T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
PspOMI  (1224)
1 site		 G G G C C C C C C G G G
ApaI  (1228)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
EcoO109I  (1258)
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.
BseRI  (1456)
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.
NsiI  (1468)
1 site		 A T G C A T T A C G T A
XbaI  (1717)
1 site		 T C T A G A A G A T C T
FseI  (1736)
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.
HpaI  (1877)
1 site		 G T T A A C C A A T T G
BsaBI  (1978)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BamHI  (1979)
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.
SalI  (1985)
1 site		 G T C G A C C A G C T G
PshAI  (2050)
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.
BspQI  (2119)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (2119)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PciI  (2235)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
NspI  (2239)
1 site		 R C A T G Y Y G T A C R
RVprimer3
20-mer  /  50% GC
1 binding site
4735 .. 4754  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
2036 .. 2055  =  20 annealed bases
Tm  =  62°C
CBG99luc
88 .. 1716  =  1629 bp
542 amino acids  =  60.1 kDa
Product: green-emitting variant of click beetle luciferase
human codon-optimized
CBG99luc
88 .. 1716  =  1629 bp
542 amino acids  =  60.1 kDa
Product: green-emitting variant of click beetle luciferase
human codon-optimized
AmpR
3055 .. 3915  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3055 .. 3846  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3055 .. 3915  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3847 .. 3915  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3055 .. 3915  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2296 .. 2884  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2296 .. 2884  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4047 .. 4502  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4047 .. 4502  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 poly(A) signal
1756 .. 1877  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1756 .. 1877  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3916 .. 4020  =  105 bp
AmpR promoter
3916 .. 4020  =  105 bp
pause site
4695 .. 4786  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
4695 .. 4786  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
MCS
1 .. 58  =  58 bp
multiple cloning site
MCS
1 .. 58  =  58 bp
multiple cloning site
poly(A) signal
4633 .. 4681  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4633 .. 4681  =  49 bp
synthetic polyadenylation signal
ORF:  88 .. 1716  =  1629 bp
ORF:  542 amino acids  =  60.1 kDa
ORF:  3185 .. 3451  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4580 .. 113  =  327 bp
ORF:  108 amino acids  =  12.9 kDa
ORF:  1842 .. 2084  =  243 bp
ORF:  80 amino acids  =  9.0 kDa
ORF:  1000 .. 1602  =  603 bp
ORF:  200 amino acids  =  22.4 kDa
ORF:  3055 .. 3915  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

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pGL4.26[luc2 minP Hygro]
pGLuc Mini-TK 2
pSF-CMV-EMCV-FLuc
pCBG99-Basic
pGL4.27[luc2P minP Hygro]
pGLuc-Basic 2
pSF-CMV-EMCV-RLuc
pCBG99-Control
pGL4.28[luc2CP minP Hygro]
phRG-B
pSF-CMV-FLuc
pCBR-Basic
pGL4.29[luc2P CRE Hygro]
phRG-TK
pSF-CMV-FMDV-FLuc
pCBR-Control
pGL4.30[luc2P NFAT-RE Hygro]
phRL-CMV
pSF-CMV-FMDV-RLuc
pCLuc Mini-TK 2
pGL4.31[luc2P GAL4UAS Hygro]
phRL-null
pSF-CMV-PGK-FLuc
pCLuc-Basic 2
pGL4.32[luc2P NF-kB-RE Hygro]
phRL-SV40
pSF-CMV-PGK-RLuc
pCMV-CLuc 2
pGL4.33[luc2P SRE Hygro]
phRL-TK
pSF-CMV-RLuc
pCMV-Cypridina Luc
pGL4.34[luc2P SRF-RE Hygro]
phRL-TK(Int-)
pSF-CMV-RSV-FLuc AscI
pCMV-Gaussia Luc
pGL4.35[luc2P 9XGAL4UAS Hygro]
pLightSwitch_Prom
pSF-CMV-RSV-RLuc AscI
pCMV-Gaussia-Dura Luc
pGL4.36[luc2P MMTV Hygro]
pLVX-MetLuc Control
pSF-CMV-Ub-FLuc AscI
pCMV-GLuc 2
pGL4.37[luc2P ARE Hygro]
pLVX-MetLuc Reporter
pSF-CMV-Ub-RLuc AscI
pCMV-Green Renilla Luc
pGL4.38[luc2P p53 RE Hygro]
pMCS-Cypridina Luc
pSF-CMVe-FLuc
pCMV-Red Firefly Luc
pGL4.39[luc2P ATF6 RE Hygro]
pMCS-Gaussia Luc
pSF-CMVe-RLuc
pCRE-MetLuc2-Reporter
pGL4.40[luc2P MRE Hygro]
pMCS-Gaussia-Dura Luc
pSF-MinProm-FLuc
pFN26A (BIND) hRluc-neo Flexi Vector
pGL4.41[luc2P HSE Hygro]
pMCS-Green Renilla Luc
pSF-MinProm-RLuc
pGEM-luc
pGL4.42[luc2P HRE Hygro]
pMCS-Red Firefly Luc
pSF-pA-CMVe-FLuc
pGL2-Basic
pGL4.43[luc2P XRE Hygro]
pMetLuc-Mem Control
pSF-pA-CMVe-RLuc
pGL2-Control
pGL4.44[luc2P AP1 RE Hygro]
pMetLuc-Mem Reporter
pSF-pA-MinProm-FLuc
pGL2-Enhancer
pGL4.45[luc2P ISRE Hygro]
pMetLuc2-Control
pSF-pA-MinProm-RLuc
pGL2-Promoter
pGL4.47[luc2P SIE Hygro]
pMetLuc2-Reporter
pSF-pA-PromMCS-FLuc
pGL3-Basic
pGL4.48[luc2P SBE Hygro]
pmirGLO
pSF-pA-PromMCS-RLuc
pGL3-Control
pGL4.49[luc2P TCF-LEF RE Hygro]
pMirTarget
pSF-PromMCS-FLuc
pGL3-Enhancer
pGL4.50[luc2 CMV Hygro]
pNFkB-MetLuc2-Reporter
pSF-PromMCS-RLuc
pGL3-Promoter
pGL4.51[luc2 CMV Neo]
pNL1.1.CMV[Nluc CMV]
pSF-RecA-FLuc
pGL4.10[luc2]
pGL4.52[luc2P STAT5 RE Hygro]
pNL1.1[Nluc]
pSP-luc+NF
pGL4.11[luc2P]
pGL4.70[hRluc]
pNL1.2[NlucP]
pSV40-CLuc
pGL4.12[luc2CP]
pGL4.71[hRlucP]
pNL1.3.CMV[secNluc CMV]
pSV40-GLuc
pGL4.13[luc2 SV40]
pGL4.72[hRlucCP]
pNL1.3[secNluc]
pTK-CLuc
pGL4.14[luc2 Hygro]
pGL4.73[hRluc SV40]
pNL2.1[Nluc Hygro]
pTK-Cypridina Luc
pGL4.15[luc2P Hygro]
pGL4.74[hRluc TK]
pNL2.2[NlucP Hygro]
pTK-Gaussia Luc
pGL4.16[luc2CP Hygro]
pGL4.75[hRluc CMV]
pNL2.3[secNluc Hygro]
pTK-Gaussia-Dura Luc
pGL4.17[luc2 Neo]
pGL4.76[hRluc Hygro]
pNL3.1[Nluc minP]
pTK-GLuc
pGL4.18[luc2P Neo]
pGL4.77[hRlucP Hygro]
pNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]
pTK-Green Renilla Luc
pGL4.19[luc2CP Neo]
pGL4.78[hRlucCP Hygro]
pNL3.2[NlucP minP]
pTK-Red Firefly Luc
pGL4.20[luc2 Puro]
pGL4.79[hRluc Neo]
pNL3.3[secNluc minP]
pXPG
pGL4.21[luc2P Puro]
pGL4.80[hRlucP Neo]
pRL-CMV

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