Contents > Transposomics™ > EZ-Tn5™ Transposon Construction Vectors
EZ-Tn5™ Transposon Construction Vectors
Supplier: EPICENTRE® Biotechnologies
EZ-Tn5™ pMOD™-2<MCS> Transposon Construction Vector
| Catalogue number | Pack size |
| MOD0602 | 20µg |
EZ-Tn5™ pMOD™-3<R6Kγori /MCS>Transposon Construction Vector
| Catalogue number | Pack size |
| MOD1503 | 20µg |
EZ-Tn5™ pMOD™-4 <MCS> Transposon Construction Vector
| Catalogue number | Pack size |
| MOD4804 | 20µg |
EZ-Tn5™pMOD™-5 <R6K γori / MCS> Transposon Construction Vector
| Catalogue number | Pack size |
| MOD4805 | 20µg |
pMOD™<MCS> Forward PCR Primer
| Catalogue number | Pack size |
| MODFP931 | 1nmole |
pMOD™<MCS> Forward Sequencing Primer
| Catalogue number | Pack size |
| MODFSP201 | 1nmole |
pMOD™<MCS> Reverse PCR Primer
| Catalogue number | Pack size |
| MODRP941 | 1nmole |
pMOD™<MCS> Reverse Sequencing Primer
| Catalogue number | Pack size |
| MODRSP202 | 1nmole |
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Last modified: 2008-07-15 14:57:15
Epicentre® offers four different EZ-Tn5™ pMOD™ Transposon Construction Vectors for the preparation of custom EZ-Tn5™ Transposons (Table 1). Each vector contains a multiple cloning site (MCS) flanked by the hyperactive 19-bp Mosaic Ends (ME, denoted by < >) that are specifically and uniquely recognized by EZ-Tn5™ Transposase. To prepare your transposon, clone any DNA sequence of interest (e.g., selectable marker, control element, reporter gene) into the MCS and then generate the transposon either by PCR amplification or restriction enzyme digestion. Any of the four Transposon Construction Vectors can be used to generate an EZ-Tn5 Transposon, but they offer different features.
The Transposon Construction Vectors pMOD-2<MCS> and pMOD-3 <R6Kγori/MCS> are pUC-based vectors. They consist of ME sequences that flank an MCS in a vector with a colE1 origin of replication (Figure 1). EZ-Tn5 pMOD-3<R6Kγori/MCS> also contains an R6Kγori within the ME sequences, which is useful for a variety of rescue cloning applications. These vectors work well for constructing transposons in most cases. However, if the transposon is prepared by restriction enzyme digestion, there is a chance that the uncut pMOD vector will contaminate the transposon constructed.
To solve this problem, the colE1 ori was eliminated in pMOD-4<MCS> and pMOD-5<R6Kγori/MCS>, which only have the R6Kγori (Figure 2). Replication from the R6Kγ origin in these two new vectors is dependent on the pir gene product produced by TransforMAX™ EC100D™ pir+ and pir-116 E.coli cells. Since most bacterial strains do not contain a pir gene, the uncut plasmid DNA that contaminates these transposon preparations can't replicate and background problems are eliminated.
| EZ-Tn5™ Transposon Construction Vectors | ori that is located on vector outside of the ME sequences | ori that is located within the ME sequences |
|---|---|---|
| pMOD-2<MCS> | colE1 | None |
| pMOD-3<R6Kγori/MCS> | colE1 | R6Kγori |
| pMOD-4<MCS> | R6Kγori | None |
| pMOD-5<R6Kγori/MCS> | None | R6Kγori |
 | Figure 1. EZ-Tn5™ Transposon Construction Vectors pMOD™-2<MCS> and pMOD™-3<R6Kγori/MCS> replicate in standard E. coli strains using a colE1 origin of replication. Transposons made with the pMOD™-3<R6Kγori/MCS> vector also have an R6Kγori within the transposon, for rescue cloning applications. |
 | Figure 2. EZ-Tn5™ Transposon Construction Vectors pMOD™-4<MCS> and pMOD™-5<R6Kγori/MCS> lack a colE1 origin of replication and require E. coli strains that produce a pir gene product for replication. This results in less background from uncut plasmid DNA when an artificial transposon is prepared by restriction endonuclease digestion. |
Last modified: 2008-05-14 16:46:46
Benefits
- The multiple cloning site (MCS) enables easy cloning of any DNA of interest
- Hyperactive 19-bp Mosaic End sequences flanking the MCS for high efficiency transposition using EZ::TN Transposase
- Unique primer binding sites at each end of the transposon for bidirectional sequencing of the insertion site using the Forward and Reverse Sequencing Primers (available separately). No need to design your own primers
- Three options can be used to prepare an EZ-Tn5™ Transposon™ digestion with Pvu II, digestion with PshA I, or PCR amplification using the PCR amplification primers provided with the vector
- EZ-Tn5™ Transposons made with pMOD-3<R6Kγori /MCS> can be used for a variety of rescue cloning applications
Last modified: 2008-05-14 16:46:46
Protocols for: EZ-Tn5™ pMOD™ <MCS> Transposon Construction Vectors
EZ-Tn5™ pMOD™ <MCS> Transposon Construction Vectors Protocol
(catalogue number MOD0602 / MOD4804)
EZ-Tn5™ pMOD™ <R6Kγori/MCS> Transposon Construction Vectors Protocol
