Plasmid

Part:BBa_K4083000:Design

Designed by: Arsen Orazbek   Group: iGEM21_NU_Kazakhstan   (2021-09-08)
Revision as of 12:10, 19 October 2021 by Arsenorazbek (Talk | contribs) (→‎Source)


pRGPDuo2 plasmid for P. putida


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3853
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3880
    Illegal BamHI site found at 31
    Illegal XhoI site found at 2175
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 6480
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BBa_K4083000-RPGDuo2_plasmid.png

Figure 1. Illustration of RPGDuo2 plasmid.

As illustrated in Figure 1, this plasmid contains two multiple cloning sites (MCS 1 and MCS2) with two transcriptional terminators. The MCS1 and MCS2 are regulated by IPTG-inducible Ptac and aTc-inducible PtetR/tetA promoters respectively. Moreover, there are lacI and tetR dependent repression systems that are regulated by Ptac and PtetR/tetA respectively. They were integrated for metabolic engineering purposes. There is a kanamycin resistance gene (KanR). There are two origins of replication to allow replication in E. coli (colE1) and P. putida (pRO1600).

Source

Modified from pRG_Duet for Corynebacterium glutamicum Gauttam et al.

References