pGLuc-Basic 2

Mammalian promoter reporter vector encoding secreted Gaussia luciferase.

Sequence Author: New England Biolabs

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AatII (4957) SgrDI (4956) ZraI (4955) SspI (4839) ScaI (4515) TatI (4513) BglI (4155) BsaI (4096) AflIII - PciI (3142) BstZ17I (2763) BsmI (2711) PfoI (2566) BglII (12) EcoRI (20) EcoRV (32) PaeR7I - PspXI - XhoI (36) HindIII (45) Acc65I (51) KpnI (55) Eco53kI (59) SacI (61) BamHI (63) BbsI (152) NruI (181) SacII (207) NotI (635) XbaI (691) DraIII (1057) CsiI - SexAI * (1348) BseRI (1577) StuI (1580) AvrII (1581) TspMI - XmaI (1602) SmaI (1604) BclI * (1632) KasI (1790) NarI (1791) SfoI (1792) PluTI (1794) PflFI - Tth111I (1909) BssHII (2188) RsrII (2307) BstBI (2473) pGLuc-Basic 2 4958 bp
AatII  (4957)
1 site		 G A C G T C C T G C A G
SgrDI  (4956)
1 site		 C G T C G A C G G C A G C T G C
ZraI  (4955)
1 site		 G A C G T C C T G C A G
SspI  (4839)
1 site		 A A T A T T T T A T A A
ScaI  (4515)
1 site		 A G T A C T T C A T G A
TatI  (4513)
1 site		 W G T A C W W C A T G W
BglI  (4155)
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  (4096)
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.
AflIII  (3142)
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  (3142)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BstZ17I  (2763)
1 site		 G T A T A C C A T A T G
BsmI  (2711)
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  (2566)
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.
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.
BbsI  (152)
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  (181)
1 site		 T C G C G A A G C G C T
SacII  (207)
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  (635)
1 site		 G C G G C C G C C G C C G G C G
XbaI  (691)
1 site		 T C T A G A A G A T C T
DraIII  (1057)
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.
CsiI  (1348)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (1348)
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  (1577)
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  (1580)
1 site		 A G G C C T T C C G G A
AvrII  (1581)
1 site		 C C T A G G G G A T C C
TspMI  (1602)
1 site		 C C C G G G G G G C C C
XmaI  (1602)
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  (1604)
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  (1632)
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  (1790)
1 site		 G G C G C C C C G C G G
NarI  (1791)
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  (1792)
1 site		 G G C G C C C C G C G G
PluTI  (1794)
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  (1909)
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  (1909)
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  (2188)
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  (2307)
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.
BstBI  (2473)
1 site		 T T C G A A A A G C T T
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3962 .. 4753  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4754 .. 4822  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
1663 .. 2457  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
1663 .. 2457  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
3203 .. 3791  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3203 .. 3791  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hGLuc
76 .. 633  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
hGLuc
76 .. 633  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
f1 ori
824 .. 1252  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
824 .. 1252  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1266 .. 1596  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1266 .. 1596  =  331 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2631 .. 2752  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2631 .. 2752  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4823 .. 4927  =  105 bp
AmpR promoter
4823 .. 4927  =  105 bp
MCS
12 .. 68  =  57 bp
multiple cloning site
MCS
12 .. 68  =  57 bp
multiple cloning site
poly(A) signal
642 .. 690  =  49 bp
synthetic polyadenylation signal
poly(A) signal
642 .. 690  =  49 bp
synthetic polyadenylation signal
SV40 ori
1447 .. 1582  =  136 bp
SV40 origin of replication
SV40 ori
1447 .. 1582  =  136 bp
SV40 origin of replication
ORF:  76 .. 633  =  558 bp
ORF:  185 amino acids  =  19.9 kDa
ORF:  775 .. 1041  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1663 .. 2457  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1835 .. 2221  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4092 .. 4358  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4898 .. 344  =  405 bp
ORF:  134 amino acids  =  14.3 kDa
ORF:  3962 .. 4822  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4719 .. 753  =  993 bp
ORF:  330 amino acids  =  36.5 kDa
ORF:  1972 .. 2508  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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Individual Sequences & Maps

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pRL-TK
pCBG68-Basic
pGL4.25[luc2CP minP]
pGL4.84[hRlucCP Puro]
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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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