pGlow-TOPO (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products to measure promoter activity using GFP.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspGlow-TOPO (linearized)
AatII (5216) SgrDI (5215) ZraI (5214) SspI (5098) ScaI (4774) PvuI (4664) BglI (4414) AhdI (4294) AlwNI (3817) PciI (3401) BsmI (2970) PfoI (2825) RsrII (2566) BssHII (2447) AflII (5279) Acc65I (5288) KpnI (5292) SpeI (5306) End (5333) Start (1) BspEI * (248) NdeI (256) BsrGI (302) MluI (348) MfeI (586) NotI (764) EcoO109I - PspOMI (782) ApaI (786) BbsI (1002) DraIII (1316) CsiI - SexAI * (1607) BseRI (1836) StuI (1839) AvrII (1840) TspMI - XmaI (1861) SmaI (1863) BsaBI * (1909) KasI (2049) NarI (2050) SfoI (2051) PluTI (2053) PflFI - Tth111I (2168) pGlow-TOPO® 5332 bp
AatII  (5216)
1 site		 G A C G T C C T G C A G
SgrDI  (5215)
1 site		 C G T C G A C G G C A G C T G C
ZraI  (5214)
1 site		 G A C G T C C T G C A G
SspI  (5098)
1 site		 A A T A T T T T A T A A
ScaI  (4774)
1 site		 A G T A C T T C A T G A
PvuI  (4664)
1 site		 C G A T C G G C T A G C
BglI  (4414)
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.
AhdI  (4294)
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  (3817)
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.
PciI  (3401)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BsmI  (2970)
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  (2825)
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.
RsrII  (2566)
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  (2447)
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.
AflII  (5279)
1 site		 C T T A A G G A A T T C
Acc65I  (5288)
1 site		 G G T A C C C C A T G G
KpnI  (5292)
1 site		 G G T A C C C C A T G G
SpeI  (5306)
1 site		 A C T A G T T G A T C A
End  (5333)
0 sites
Start  (1)
0 sites
BspEI  (248)
1 site		 T C C G G A A G G C C T
* Blocked by Dam methylation.
NdeI  (256)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BsrGI  (302)
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.
MluI  (348)
1 site		 A C G C G T T G C G C A
MfeI  (586)
1 site		 C A A T T G G T T A A C
NotI  (764)
1 site		 G C G G C C G C C G C C G G C G
EcoO109I  (782)
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.
PspOMI  (782)
1 site		 G G G C C C C C C G G G
ApaI  (786)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BbsI  (1002)
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.
DraIII  (1316)
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  (1607)
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  (1607)
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  (1836)
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  (1839)
1 site		 A G G C C T T C C G G A
AvrII  (1840)
1 site		 C C T A G G G G A T C C
TspMI  (1861)
1 site		 C C C G G G G G G C C C
XmaI  (1861)
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  (1863)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BsaBI  (1909)
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.
KasI  (2049)
1 site		 G G C G C C C C G C G G
NarI  (2050)
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  (2051)
1 site		 G G C G C C C C G C G G
PluTI  (2053)
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  (2168)
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  (2168)
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.
AmpR
4221 .. 5081  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4221 .. 5012  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4221 .. 5081  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5013 .. 5081  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4221 .. 5081  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
1922 .. 2716  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1922 .. 2716  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
Cycle 3 GFP
24 .. 743  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   24 .. 26  =  3 bp
   1 amino acid  =  149.2 Da
Product: Cycle 3 GFP (Crameri et al., 1996)
mammalian codon-optimized
Cycle 3 GFP
24 .. 743  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   27 .. 29  =  3 bp
   1 amino acid  =  89.1 Da
Product: Cycle 3 GFP (Crameri et al., 1996)
mammalian codon-optimized
Cycle 3 GFP
24 .. 743  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   30 .. 743  =  714 bp
   237 amino acids  =  26.7 kDa
Product: Cycle 3 GFP (Crameri et al., 1996)
mammalian codon-optimized
Cycle 3 GFP
24 .. 743  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Cycle 3 GFP (Crameri et al., 1996)
mammalian codon-optimized
ori
3462 .. 4050  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3462 .. 4050  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1083 .. 1511  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1083 .. 1511  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1525 .. 1855  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1525 .. 1855  =  331 bp
SV40 enhancer and early promoter
bGH poly(A) signal
813 .. 1037  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
813 .. 1037  =  225 bp
bovine growth hormone polyadenylation signal
SV40 poly(A) signal
2890 .. 3011  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2890 .. 3011  =  122 bp
SV40 polyadenylation signal
AmpR promoter
5082 .. 5186  =  105 bp
AmpR promoter
5082 .. 5186  =  105 bp
T7 promoter
5234 .. 5252  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5234 .. 5252  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
1706 .. 1841  =  136 bp
SV40 origin of replication
SV40 ori
1706 .. 1841  =  136 bp
SV40 origin of replication
ORF:  4351 .. 4617  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1034 .. 1300  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1922 .. 2716  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3 .. 743  =  741 bp
ORF:  246 amino acids  =  27.7 kDa  (no start codon)
ORF:  2094 .. 2480  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4221 .. 5081  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2231 .. 2767  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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Individual Sequences & Maps

Allele TA Vector (linearized)
pCR2.1-TOPO (linearized)
pJAZZ-OK GC (left arm)
pSpark TA
pBAD-TOPO (linearized)
pCR4-TOPO (linearized)
pJAZZ-OK GC (right arm)
pSpark TA Done
pBAD Thio-TOPO (linearized)
pCR8 GW TOPO (linearized)
pLenti6.3 V5-TOPO (linearized)
pSTBlue-1 AccepTor
pBlue-TOPO (linearized)
pCRII-TOPO (linearized)
pLenti7.3 V5-TOPO (linearized)
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pcDNA3.1 CT-GFP-TOPO (linearized)
pDrive (linearized)
pLPS-T (linearized)
pTA Plus (linearized)
pcDNA3.1 NT-GFP-TOPO (linearized)
pEF6 V5-His-TOPO (linearized)
pLUG (linearized)
pTargeT (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pENTR5'-TOPO (linearized)
pLUG-Multi (linearized)
pTOP TA V2 (linearized)
pcDNA3.3-TOPO (linearized)
pET SUMO (linearized)
pMT V5-His-TOPO (linearized)
pTrcHis-TOPO (linearized)
pcDNA3.4-TOPO (linearized)
pETBlue-1 AccepTor
pOptiVEC-TOPO (linearized)
pTrcHis2-TOPO (linearized)
pcDNA4 HisMax-TOPO (linearized)
pEXP5-CT TOPO (linearized)
pPrime (linearized)
pTZ57R_T
pcDNA5 FRT TO-TOPO (linearized)
pEXP5-NT TOPO (linearized)
pQE-30 UA (linearized)
pUC57-T
pcDNA5 FRT V5-His-TOPO (linearized)
pGC Blue (linearized)
pSC-A-amp kan (linearized)
pYES2.1 V5-His-TOPO (linearized)
pcDNA6.2 C-EmGFP-GW TOPO (linearized)
pGEM-T (linearized)
pSecTag FRT V5-His-TOPO (linearized)
RBC T&A Cloning Vector (linearized)
pcDNA6.2 C-YFP-GW TOPO (linearized)
pGEM-T Easy (linearized)
pSMART GC HK (linearized)
T-Vector pMD19 (Simple)
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pGlow-TOPO (linearized)
pSMART GC LK (linearized)
T-Vector pMD20
pCR-XL-TOPO (linearized)
pIB V5-His-TOPO (linearized)

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
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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