Entry Clones

An Entry clone contains your gene of interest flanked by attL sequences, which are then used to recombine with attR sequences to create your desired expression clone. There are three methods you can use to produce an Entry clone: BP cloning, restriction enzyme and ligase cloning, and TOPO® cloning into a Gateway® Entry vector, which is the most common method. 

Selection Guide for Entry Vectors

Featured Products

Gateway® Entry Options

  • RNAi Expression - Efficient delivery and long-term stable or transient shRNA expression in any mammalian cell types with Lentiviral and Adenoviral Gateway® vectors
  • Viral Expression - High level, stable and transient gene expression in any mammalian cell type
  • In Vitro Protein Synthesis - Directly synthesize high yields of recombinant protein in a single reaction tube in just 2 hours without special equipment. Scale the reaction to achieve microgram to milligram levels of protein
  • In Vivo Biotinylation - Easy efficient method for expressing, purifying and detecting biotinylated recombinant proteins.
  • Tag-On-Demand ™ Technology - Produce native or tagged proteins from one vector.

Ordering Information

Catalog # Name Size List Price (USD) Qty
12535029 PCR Cloning System with Gateway® Technology with pDONR™221 & OmniMAX™2 Competent Cells 20 reactions 394.00
12535037 PCR Cloning System with Gateway® Technology with pDONR™/Zeo & OmniMAX™2 Competent Cells 20 reactions 476.00
12536017 Gateway® pDONR™221 Vector 6 µg 204.00
12535035 Gateway® pDONR™/Zeo Vector 6 µg 339.00
12537023 MultiSite Gateway® Three-Fragment Vector Construction Kit 20 reactions 2,165.00
12537102 MultiSite Gateway® Pro 2.0, for cloning two DNA fragments into a Gateway® destination vector 20 reactions 1,834.00
12537103 MultiSite Gateway® Pro 3.0, for cloning three DNA fragments into a Gateway® destination vector 20 reactions 2,165.00
12537104 MultiSite Gateway® Pro 4.0, for cloning four DNA fragments into a Gateway® destination vector 20 reactions 2,455.00
12537100 MultiSite Gateway® Pro Plus, for flexible cloning of up to four DNA fragments into a Gateway® destination vector 20 reactions 2,655.00
12537162 pcDNA™6.2/V5-PL-DEST Mammalian Expression Vector 6 µg 416.00
K16001 Champion™ pET160 Directional TOPO® Expression Kit with Lumio™ Technology 20 reactions 784.00
K16101 Champion™ pET161 Directional TOPO® Expression Kit with Lumio™ Technology 20 reactions 794.00
K244020 pcDNA™3.2/GW/D-TOPO® Expression Kit 20 reactions 652.00
K246020 pcDNA™6.2/GW/D-TOPO® Expression Kit 20 reactions 654.00
12578084 GeneBLAzer® C-terminal TOPO® Fusion Kit, for in vivo detection 1 kit 800.00
12578076 GeneBLAzer® C-terminal TOPO® Fusion Kit, for in vitro detection 1 kit 800.00
12578100 GeneBLAzer® N-terminal TOPO® Fusion Kit, for in vivo detection 1 kit 800.00
K35920 Vivid Colors™ pcDNA™6.2/C-EmGFP-GW/TOPO® Mammalian Expression Vector 20 reactions 1,440.00
K36120 Vivid Colors™ pcDNA™6.2/C-YFP-GW/TOPO® Mammalian Expression Vector 20 reactions 1,458.00
K36020 Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector 20 reactions 1,434.00

Building an Entry Clone

The three possible methods that lead to the Entry clone are depicted below: A. BP cloning, B. TOPO® cloning and C. restriction enzyme and ligase cloning. Red arrows represent the fragment of interest. Adapted from Katzen, F. 2007) Expert Opin Drug Discov 2(4): 571–589.

pENTR™/D-TOPO® Vector Kits

pENTR/D-TOPO® vectors take advantage of fast, efficient Directional TOPO® cloning that delivers your insert in the correct orientation for expression. These vectors contain the necessary attL sequences for recombination into any Destination vector and certain versions carry a TEV protease cleavage site for producing native proteins after expression (Figure 1).

Figure 1 - pENTR/D-TOPO

Figure 1. Several pENTR™ vectors are available for Directional TOPO® cloning and direct access to the multitude of Gateway® expression vectors.

pCR®8/GW/TOPO® Vector Kits

The pCR®8/GW/TOPO® vector enables efficient TOPO®-TA cloning. These vectors easily facilitate multipurpose use: rapid recombination into a variety of Gateway® Destination vectors, convenient sequencing, robust selection in E. coli with spectinomycin resistance, and easy exicision of insert DNA with flanking EcoR I sites (Figure 2).

pCR8/GW/TOPO


Figure 2. The pCR®8/GW/TOPO® Entry vector allows TOPO® TA Cloning® for multiple downstream applications.

Selection Guide for Entry Vectors

Rating Kit SKU Advantages
Best: high efficiency, directional TOPO® Cloning Vector KitspENTR™/TEV/D-TOPO® Cloning KitK253520
  • Directional TOPO® Cloning
  • 5’ TEV sequence for N-terminal tag removal
  • Includes fast-growing competent E. coli that shortens the time for miniprep
 pENTR™/D-TOPO® Cloning KitK240020
  • Directional TOPO® Cloning
 pENTR™/SD/D-TOPO® Cloning KitK242020
  • Directional TOPO® Cloning
  • Vector includes the Shine-Dalgarno sequence for E. coli expression-ready Entry clone
 pENTR™/TEV/D-TOPO® Cloning KitK252520
  • Directional TOPO® Cloning
  • 5’ TEV sequence for N-terminal tag removal
Very good: high efficiency TOPO® TA Cloning Vector KitspCR®8/GW/TOPO® TA Cloning kitK252002
  • Efficient TOPO® TA cloning kitIncludes fast-growing competent E. coli that shortens the time for miniprep
  • Includes plasmid purification components for maximum convenience
 pCR®8/GW/TOPO® TA cloning kitK252020
  • Efficient TOPO® TA cloning with fast-growing competent E. coli that shortens
  • Includes fast-growing competent E. coli that shortens the time for miniprep
 pCR®8/GW/TOPO® TA Cloning KitK250020
  • Efficient TOPO® TA cloning simplifies Entry clone construction
Standard restriction cloning VectorspENTR™ 1A Dual Selection VectorA10462
  • Restriction enzyme cloning vector that produces in-frame (rf = 0), expression ready Entry clones, including both Shine-Dalgarno and Kozak sequences
 pENTR™ 2B Dual Selection VectorA10463
  • Restriction enzyme cloning vector that produce in-frame (rf = +1), expression ready Entry clone
  • Chloramphenical and Kanamycin selection
 pENTR™ 3C Dual Selection VectorA10464
  • Restriction enzyme cloning vector that produce in-frame (rf = +2), expression ready Entry clones
  • Chloramphenical and Kanamycin selection
 pENTR™ 4 Dual Selection VectorA10465
  • Same as pENTR™ 1A Vector except with Nco I instead of Dra I in MCS that produces in-frame (rf = 0), expression-ready Entry clones
  • Chloramphenical and Kanamycin selection
 pENTR™ 11 Dual Selection VectorA10467
  • Same as pENTR™ 1A Vector except with Nsp V instead of Dra I in MCS that produces in-frame (rf = 0), expression-ready Entry clones
  • Chloramphenical and Kanamycin selection

Ordering Information

Catalog # Name Size List Price (USD) Qty
K250020 pCR®8/GW/TOPO® TA Cloning Kit with One Shot® TOP10 E. coli 20 reactions 395.00
K252002 pCR®8/GW/TOPO® TA Cloning Kit with One Shot® Mach1™-T1R E. coli and PureLink® Kit 20 preps 485.00
K252020 pCR®8/GW/TOPO® TA Cloning Kit with One Shot® Mach1™-T1R E. coli 20 reactions 395.00
K252020SC pCR®8/GW/TOPO® TA Cloning Kit with One Shot® Mach1™-T1R Chemically Competent E. coli (Supply Center Only) 20 reactions 398.00
K250020SC pCR®8/GW/TOPO® TA Cloning Kit with One Shot® TOP10 Chemically Competent E. coli (Supply Center Only) 20 reactions 398.00
A10467 Gateway® pENTR™ 11 Dual Selection Vector 10 µg 183.00
A10462 Gateway® pENTR™ 1A Dual Selection Vector 10 µg 183.00
A10463 Gateway® pENTR™ 2B Dual Selection Vector 10 µg 183.00
A10464 Gateway® pENTR™ 3C Dual Selection Vector 10 µg 183.00
A10465 Gateway® pENTR™ 4 Dual Selection Vector 10 µg 183.00
K59120 pENTR™ 5'-TOPO® TA Cloning® Kit, with One Shot® TOP10 Chemically Competent E. coli 20 reactions 422.00
K243520 pENTR™/D-TOPO® Cloning Kit, with One Shot® Mach1™-T1R Chemically Competent E. coli 20 reactions 398.00
K240020 pENTR™/D-TOPO® Cloning Kit, with One Shot® TOP10 Chemically Competent E. coli 20 reactions 403.00
K263520 pENTR™/SD/D-TOPO® Cloning Kit, with One Shot® Mach1™-T1R Chemically Competent E. coli 20 reactions 408.00
K242020 pENTR™/SD/D-TOPO® Cloning Kit, with One Shot® TOP10 Chemically Competent E. coli 20 reactions 395.00
K253520 pENTR™/TEV/D-TOPO® Cloning Kit, with One Shot® Mach1™-T1R Chemically Competent E. coli 20 reactions 408.00
K252520 pENTR™/TEV/D-TOPO® Cloning Kit, with One Shot® TOP10 Chemically Competent E. coli 20 reactions 395.00