Plasmid name: pUHC 13-6 Size: ca.6500 bp Resistance: Ampicillin constructed by: Manfred Gossen Host strain: E.coli "sure" Reference: Manfred Gossen unpublished
pUHC13-6 is a derivative of pT81 luc (ref.: Nordeen, S.K. 1988 BioTechniques 6: 454-457) which contains the luciferase gene under control of the Tk promoter (position -81 to +52). For details on structure and construction of pT81 luc, please check the above reference. To bring the luciferase gene under tTA control, a heptamerized tet operator sequence and a polylinker were inserted upstream of the Tk promoter (see attached sequence).
The plasmid consists of three main fragments:
SalI SmaI KpnI SstI XhoI GTCGACCCGGGTACCGAGCTC ( GACTTTCACTTTTCTCTATCACTGATAGGGAGTGGTAAACTC ) GAGAT 7 CCGGCGAATTCGAACACGCAGATGCAGTCGGGGCGGCGCGGTCCGAGGTCCACTTCGCATATTAAGGTG -81 -> ACGCGTGTGGCCTCGAACACCGAG +1
* the Tk minimal promoter region with the heptamerized tet operators is shown: