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Revision as of 01:27, 28 September 2013


PART DESCRIPTION:Constitutive promoter

This is a medium-strength Escherichia coli Ptrc* promoter. The asterisk denotes the lack of an operator downstream of the transcription start site, -35 to +1 of the promoter. The promoter's sequence comes from "Precise and reliable gene expression via standard transcription and translation initiation elements" by Vivek K. Mutalik and colleagues. Two inward pointing BpiI (BbsI) sites were added between the original promoter sequence and the BioBrick prefix and suffix to allow seamless assembly into the 2013 Purdue iGEM team's Bicistronic Design (BCD) Expression Operating Units, modified versions of BCDs 7, 18, 22 and 23 created by Mutalik et al. in the aforementioned work.


Contact Information


  • Author(s): James Nolan and Chris Thompson
  • Team: Purdue University 2013
  • Data Collection: Amanda Shanley, Chris Thompson, and James Nolan
  • Affiliation: Purdue University (Bindley Bioscience Center)
  • Contact: nolan6@purdue.edu
  • Related Parts: None
  • Date: 09/27/13

Standard Design Information


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]


  • Chassis: E.coli
  • Strain: BL21
  • Device Name: BBa_K1225000
  • Device Type: Promoter
  • Safety Level: Risk Group 1
  • Assembly: Golden Gate Assembly
  • Protocol: Freiburg Golden Gate Protocol
  • Scars: None
  • Insertion: Plasmid
  • Vector: pSB1C3

Growth Rate Assay


BASIC INFORMATION

Purpose: To assess what effect, if any, our genetic parts have on the growth rate of E.coli.

Chassis: E.coli

Strain: BL21

Protocols: Purdue iGEM Growth Curve Protocol

Date: 09/25/13


GROWTH CONDITIONS

Media Type: Luria Broth (LB)

Vessel: 10mL Culture Tube

Volume: 5mL

Incubation: 37 C, 250 rpm


MEASUREMENT INFORMATION

Data Type: Growth Curve (OD vs Time)

Location: Bindley Bioscience Center

Machine Name: N/A

Time Interval: 30min

Total Time: 270min


Proof of Functionality


BASIC INFORMATION

Purpose: To qualitatively and quantitatively prove that our promoter initiates transcription by placing mRFP after it.

Chassis: E.coli

Strain: BL21

Protocols: None

Date: 09/26/13


GROWTH CONDITIONS

Media Type: Luria Broth (LB)

Vessel: 10mL Culture Tube

Volume: 5mL

Incubation:37 C, 250 rpm


MEASUREMENT INFORMATION

Data Type: Fluorescence Intensity vs Wavelength

Location: Bindley Bioscience Center

Machine Name: N/A

Time Interval: 30min

Total Time: 270min