PTRIPLESCRIPT(TM) - A NOVEL CLONING VECTOR FOR GENERATING IN-VITRO TRANSCRIPTS FROM TANDEM PROMOTERS FOR SP6, T7 AND T3 RNA-POLYMERASES

We have constructed a family of novel in vitro transcription vectors i n which functional T3, T7 and SP6 RNA polymerase promoters are arrange d in tandem and directed towards a multiple cloning site. This prototy pe vector named pTRIPLEscript(TM), permits the transcription of one st rand of a DNA insert by any of the three commonly used bacteriophage R NA polymerases with no apparent cross talk, i.e., use of the wrong pro r,moter sequence. The vector has two main uses: (i) to clone probe seq uences that will be distributed to many laboratories, allowing the use of the most convenient RNA polymerase: and (ii) to circumvent the pro blem of RNA polymerase-dependent premature termination.