Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jama Hagi-Yusuf
Date created: 2013-09-12 11:00:00
Date modified: 2015-05-08 01:08:50
PhiC31 integrase and PhiC31 RDF under lock and key control with lacI Expression Cassette
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1039022_sequence (Version 1) |
Description
PhiC31 integrase (Bba_K1039012) and lacI repressor (BBa_Q04121) are under the constitutive strong promoter BBa_J23119. Key (BBa_I714036) and PhiC31 RDF (Bba_K1039013) are under the lac inducible promoter BBa_R0011. The transcriptional terminator BBa_B0015 was placed after each coding section (the integrase, the key, the RDF and the lacI cassette)PhiC31 integrase (Bba_K1039012), some binding sites associated with the cis-repressive RNA ???lock??? (Bba_K1039013), so translation is inhibited. The trans-activating RNA ???key??? (BBa_I714036) can displace the lock and allows translation.
Together, PhiC31 integrase and RDF (Bba_K1039012 and Bba_K1039013) catalyze site-specific recombination between PhiC31 attR and attL sites (Bba_K1039037 and Bba_K1039038)
They will also allow flipping of PhiC31 invertible promoter switches (Bba_K1039010 and Bba_K1039011) from RL state to PB state.
Lock and key regulate PhiC31 integrase and PhiC31 RDF expression. PhiC31 integrase and PhiC31 RDF are regulated by lock. Key expression is controlled by a lacI repressor, upon IPTG addition to the media, key will be expressed, which then unlocks PhiC31 integrase and PhiC31 RDF.
Notes
For PhiC31 RDF, a base was added after the stop codon to prevent creation of an SpeI site due to addition of suffix. For PhiC31 integrase, there was a silent mutation on base 52; changed from T to C.Source
The parts to this sequenced were either synthesized by BioBasic Inc, or from the iGEM distribution kit.| Access | Instance | Definition |
| public public public public public public public public public public public | BBa_J23119 BBa_Q04121 BBa_J01010 BBa_K1039012 BBa_B0015 BBa_R0011 BBa_I714036 BBa_B0015 BBa_R0011 BBa_J01010 BBa_K1039013 | BBa_J23119 lacI Lock 1 BBa_K1039012 BBa_B0015 lacI+pL BBa_I714036 BBa_B0015 lacI+pL Lock 1 BBa_K1039013 |