From 2007.igem.org

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Contents 1 Preliminaries 1.1 SopII 1.2 HtrII 1.3 SopII/HtrII fusion 1.4 ComP 1.5 ComA 2 Chimera Design 2.1 Overview 2.2 SopII Analysis 2.3 HtrII Analysis 2.4 ComP Analysis 2.5 Chosen Fusion

Preliminaries

SopII

sensory rhodopsin II

DNA, AA, [http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&cmd=search&term=sopII genbank]

• No IGEM restriction sites
• HaeII (@168) present

HtrII

sensory rhodopsin II transducer

DNA, AA, [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=3702851&ordinalpos=1&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum genbank]

• No IGEM restriction sites
• No useful restriction site

SopII/HtrII fusion

DNA, AA (frame 2 in Translate - only SopII + HtrII + Linker included in link), linker = TSASA SNGASA,

[http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&cmd=search&term=sopII genbank]

• No IGEM restriction sites
• Use site from SopII (this is the one that will be used)

ComP

two-component sensor histidine kinase

DNA (includes IGEM forward and reverse primers (VF2 + VR) and IGEM prefix and suffix, AA, [http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&cmd=search&term=sopII genbank]

SpeI (@927) deleted HaeII (@1902) inserted

ComA

sensory rhodopsin II transducer

DNA, AA, [http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&cmd=search&term=sopII genbank]

SpeI (@12) deleted HaeII (@433) inserted

Chimera Design

Overview

Refer to 4 PCR primer method here, 2001 paper

SopII Analysis

transmembrane photoreceptor, requires all-trans retinal as substrate... linked to HtrII insert TM analysis here. reference the three papers cited in background

HtrII Analysis

Yeh!!!! sequence analysis.

ComP Analysis

Chosen Fusion

File:Seq align.jpg Media:Example.ogg