pGLuc Mini-TK 2

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

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) BglII (12) EcoRI (20) EcoRV (32) PaeR7I - PspXI - XhoI (36) HindIII (45) Acc65I (51) KpnI (55) Eco53kI (59) SacI (61) BamHI (63) MluI (85) BbsI (222) NruI (251) SacII (277) NotI (705) XbaI (761) DraIII (1127) CsiI - 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) BstBI (2543) 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.
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.
CsiI  (1418)
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  (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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
BstBI  (2543)
1 site		 T T C G A A A A G C T T
AmpR
4032 .. 4892  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   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
2 segments
   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
ORF:  4162 .. 4428  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  146 .. 703  =  558 bp
ORF:  185 amino acids  =  19.9 kDa
ORF:  845 .. 1111  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1733 .. 2527  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1905 .. 2291  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4789 .. 414  =  654 bp
ORF:  217 amino acids  =  24.3 kDa
ORF:  4032 .. 4892  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  128 .. 823  =  696 bp
ORF:  231 amino acids  =  25.0 kDa
ORF:  2042 .. 2578  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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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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