pCLuc Mini-TK 2

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

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) PaqCI (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).
PaqCI  (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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
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
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 T A G G G G A T C C
TspMI  (2775)
1 site		 C C C G G G G G G C C C
XmaI  (2775)
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  (2777)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
KasI  (2963)
1 site		 G G C G C C C C G C G G
NarI  (2964)
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  (2965)
1 site		 G G C G C C C C G C G G
PluTI  (2967)
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.
CLuc
145 .. 1806  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
contains 30 codon substitutions for efficient translation in mammalian cells
CLuc
145 .. 1806  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
contains 30 codon substitutions for efficient translation in mammalian cells
AmpR
5135 .. 5995  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5135 .. 5926  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5135 .. 5995  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5927 .. 5995  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5135 .. 5995  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2836 .. 3630  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2836 .. 3630  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
4376 .. 4964  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4376 .. 4964  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1997 .. 2425  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1997 .. 2425  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2439 .. 2769  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
2439 .. 2769  =  331 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
3804 .. 3925  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3804 .. 3925  =  122 bp
SV40 polyadenylation signal
AmpR promoter
5996 .. 6100  =  105 bp
AmpR promoter
5996 .. 6100  =  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
MCS
20 .. 68  =  49 bp
multiple cloning site
MCS
20 .. 68  =  49 bp
multiple cloning site
poly(A) signal
1815 .. 1863  =  49 bp
synthetic polyadenylation signal
poly(A) signal
1815 .. 1863  =  49 bp
synthetic polyadenylation signal
SV40 ori
2620 .. 2755  =  136 bp
SV40 origin of replication
SV40 ori
2620 .. 2755  =  136 bp
SV40 origin of replication
Kozak sequence
139 .. 148  =  10 bp
Kozak sequence
139 .. 148  =  10 bp
ORF:  145 .. 1806  =  1662 bp
ORF:  553 amino acids  =  61.5 kDa
ORF:  1948 .. 2214  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2836 .. 3630  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  899 .. 1210  =  312 bp
ORF:  103 amino acids  =  11.3 kDa
ORF:  3008 .. 3394  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  5265 .. 5531  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1514 .. 1738  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  5135 .. 5995  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5892 .. 75  =  315 bp
ORF:  104 amino acids  =  12.2 kDa
ORF:  1402 .. 1770  =  369 bp
ORF:  122 amino acids  =  13.8 kDa
ORF:  3145 .. 3681  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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Individual Sequences & Maps

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pGL4.81[hRlucCP Neo]
pRL-null
pBIND-ER(alpha)
pGL4.23[luc2 minP]
pGL4.82[hRluc Puro]
pRL-SV40
pBIND-GR
pGL4.24[luc2P minP]
pGL4.83[hRlucP Puro]
pRL-TK
pCBG68-Basic
pGL4.25[luc2CP minP]
pGL4.84[hRlucCP Puro]
pSELECT-zeo-Lucia
pCBG68-Control
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

Plasmid Sets

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I.M.A.G.E. Consortium Plasmids
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Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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