Pgps3 Unique Restriction Sites =link= Jun 2026

Happy cloning!

The vector is a specialized 4,293 bp plasmid primarily utilized as a "Transprimer" donor within the GPS-M Mutagenesis System . Developed by New England Biolabs (NEB) , this vector is critical for in vitro random mutagenesis, where a mobile genetic element (the Transprimer) is moved from the donor plasmid into a target DNA molecule via TnsABC transposase.

However, successful cloning depends entirely on one thing: knowing which restriction enzymes cut once —and only once—in your plasmid.

: The vector contains an ampicillin resistance gene for maintenance in E. coli . The ScaI site is located within this gene; while unique, using it for cloning will disrupt ampicillin selection. Cloning Considerations pgps3 unique restriction sites

For seamless assembly (no restriction ligation), cut PGPS3 with a like PstI or NotI . Then:

Avoid using these enzymes for cloning, as they cut the plasmid multiple times:

Just remember: no SalI, no EcoRV, and always verify your map. When in doubt, a quick virtual digest takes 30 seconds and saves three days of failed ligations. Happy cloning

A means the recognition sequence appears only once in the entire plasmid backbone. This is gold for molecular cloning because:

If "pgps3" refers to the NEB vector (used for Tn7 transposition), the unique sites are centered around the transposon arms, specifically NotI , PmeI , and SpeI . However, successful cloning depends entirely on one thing:

: The Transprimer-1 element in pGPS3 contains a kanamycin resistance ( KnRcap K n to the cap R-th power

| Enzyme | Problem | |--------|---------| | | Cuts twice (once in MCS, once in the pSa origin of replication). | | EcoRV | Cuts three times (unexpectedly in the LacZ alpha fragment and the kanamycin resistance gene). | | ClaI | Cuts twice (in the MCS and the ColE1 origin). |

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