Regulatory

Part:BBa_R0187

Designed by: Bartholomew Canton   Group: iGEM2006_MIT   (2008-04-08)


T7 promoter (lacI repressible)

A lac repressible T7 promoter. This promoter is similar to those found in the pET vectors with T7lac promoters. I mutated the -9 base to an G, which should make it less than 3% of the strength of consensus. The base numbering is as used by Studier and coworkers (JMB 1991, 219, 45-59). See BBa_R0183 for more information. Note: This part has been improved by Edinburgh_UG 2017. See their improved part here: BBa_K2406020

SHSBNU_China 2021

In this project, we used combined part of T7 and LacI binding site (BBa_R0187) in our gene circuit. This part is the regulatory region in our circuit that is responsible for the controlled expression of our proteins (BBa_K3798015, BBa_K3798033) that we have experimented with different induction intensity to obtain the best IPTG usage. To express sufficient amount of CBD-eGFP protein for our use, we applied different concentrations of IPTG to the induction process of e.coli incubation.

T--SHSBNU China--Contribution 1.png

Fig.1 The relationship between fluorescent value and IPTG concentration

As shown in the chart, the fluorescent value increases with the increased IPTG concentration. Although in the diagram the leakage of this T7 promoter seems significant, this is actually caused by the use of LB medium in our culture that interfered with the fluorescent value.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//rnap/bacteriophage/T7
//direction/forward
//chassis/prokaryote/ecoli
//promoter
//regulation/negative
//chassis/bacteriophage/T7
Parameters
negative_regulators1
positive_regulators