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FX cloning System
(Kit # 1000000039 )

Depositing Labs:   Raimund Dutzler, Eric Geertsma

The 35 plasmid FX cloning kit can be used to insert a sequence of interest into a cloning vector and easily swap into expression vectors for several organisms with a variety of N- and C-terminal tags.

This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80°C immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs

Add to Cart
$165 USD + shipping

Available to academics and nonprofits only.

Original Publication

A versatile and efficient high-throughput cloning tool for structural biology. Geertsma ER and Dutzler R. Biochemistry 2011 Apr 19;50(15):3272-8. doi: 10.1021/bi200178z. Epub 2011 Mar 25 PubMed

Description

The FX cloning system (fragment exchange cloning) is an efficient, inexpensive cloning method that utilizes a Type IIS restriction enzyme and negative selection markers. This system is suitable for both prokaryotic and eukaryotic expression systems and combines the advantages of established recombination- and ligation-independent cloning methods, while reducing undesired cloning-related sequences added in the process.

The FX cloning system uses a set of three initial cloning vectors (pINIT_cat, pINIT_kan and pINIT_tet) for the initial cloning of a PCR product into one of the vectors, which differ by bacterial resistances. The subcloning of the ORF from the pINITIAL vector into an expression vector utilizes the same cloning procedure. PCR products can also be directly cloned into expression vectors.

The FX cloning Kit contains 3 initial cloning vectors, 12 E. coli expression vectors, 3 L. lactis expression vectors, 4 P. pastoris expression vectors, 4 S. cerevisiae expression vectors, 4 mammalian expression vectors, 2 Xenopus oocyte expression vectors and 3 insect expression vectors. A variety of expression vectors are available containing several combinations of 3C protease-cleavable N-terminal and C-terminal tags , including decaHis, Myc, Streptavidin Binding Peptide and Avi tags plus GFP.

FX cloning Method Overview

As described in the original publication, a PCR product containing a sequence of interest can be cloned into either an FX sequencing vector or an FX expression vector.

Illustration of FX cloning process for a PCR product into a cloning or expression vector

FX Subcloning Overview

Once a sequence of interest has been cloned into an FX sequencing vector, it can easily be transferred into an expression vector by digestion and ligation.

Illustration of subcloning process from cloning to expression vector

Kit Documentation

When receiving kit as frozen glycerol stocks on dry ice:

Icon FX cloning kit (262.3 KB)

Resources

More information on these plasmids and their use can be found in the original publication.

The FX Cloning website has an automated primer design function to assist with the FX Cloning method.

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:
"The plasmid kit used for fragment exchange cloning was a gift from Raimund Dutzler and Eric Geertsma (Addgene kit # 1000000039)"

For your Reference section:
A versatile and efficient high-throughput cloning tool for structural biology. Geertsma ER and Dutzler R. Biochemistry 2011 Apr 19;50(15):3272-8. doi: 10.1021/bi200178z. Epub 2011 Mar 25 PubMed

FX cloning Kit - #1000000039

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Chloramphenicol and Ampicillin
Chloramphenicol and Bleocin (Zeocin)
Chloramphenicol and Kanamycin
Tetracycline
Chloramphenicol

Inventory

Well Plasmid Resistance
A / 1 pBXNH3
Chloramphenicol and Ampicillin
A / 2 pBXC3H
Chloramphenicol and Ampicillin
A / 3 pBXNH3CA
Chloramphenicol and Ampicillin
A / 4 pBXC3GH
Chloramphenicol and Ampicillin
A / 5 pBXCA3GH
Chloramphenicol and Ampicillin
A / 6 pBXCA3H
Chloramphenicol and Ampicillin
A / 7 pBXNAC3GH
Chloramphenicol and Ampicillin
A / 8 pBXNAC3H
Chloramphenicol and Ampicillin
A / 9 pBXNH3A
Chloramphenicol and Ampicillin
A / 10 pREXC3GH
Chloramphenicol and Ampicillin
A / 11 pREXC3H
Chloramphenicol and Ampicillin
A / 12 pREXNH3
Chloramphenicol and Ampicillin
B / 1 pYEXNH3
Chloramphenicol and Ampicillin
B / 2 pYEXNHG3
Chloramphenicol and Ampicillin
B / 3 pYEXC3H
Chloramphenicol and Ampicillin
B / 4 pYEXC3GH
Chloramphenicol and Ampicillin
B / 5 pcDXNSM3
Chloramphenicol and Ampicillin
B / 6 pcDXNSMG3
Chloramphenicol and Ampicillin
B / 7 pcDXC3MS
Chloramphenicol and Ampicillin
B / 8 pcDXC3GMS
Chloramphenicol and Ampicillin
B / 9 pTLNX
Chloramphenicol and Ampicillin
B / 10 pTLNXA7
Chloramphenicol and Ampicillin
B / 11 pFBXNH3
Chloramphenicol and Ampicillin
B / 12 pFBXC3H
Chloramphenicol and Ampicillin
C / 1 pFBXC3GH
Chloramphenicol and Ampicillin
C / 2 pICXNH3
Chloramphenicol and Bleocin (Zeocin)
C / 3 pICXNHG3
Chloramphenicol and Bleocin (Zeocin)
C / 4 pICXC3H
Chloramphenicol and Bleocin (Zeocin)
C / 5 pICXC3GH
Chloramphenicol and Bleocin (Zeocin)
C / 6 p7XNH3
Chloramphenicol and Kanamycin
C / 7 p7XC3H
Chloramphenicol and Kanamycin
C / 8 p7XC3GH
Chloramphenicol and Kanamycin
C / 9 pINIT_kan
Chloramphenicol and Kanamycin
C / 10 pINIT_tet
Tetracycline
C / 11 pINIT_cat
Chloramphenicol
Data calculated @ 2017-08-22

Kit Plate Map - #1000000039

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