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
Bxb1 integrase and Recombination Directionality Factor (RDF) Under ???Lock and Key??? Control with L
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1039021_sequence (Version 1) |
Description
Bxb1 integrase (BBa_K1039003) and lacI repressor (BBa_Q04121) are under the constitutive strong promoter BBa_J23119. Key (BBa_I714036) and Bxb1 RDF (BBa_K1039006) are under the lac inducible promoter BBa_R0011.Bxb1 integrase (BBa_K1039003) and recombination directionality factor (RDF) (BBa_K1039006) with ribosome binding sites associated with the cis-repressive RNA ???lock??? (BBa_J01010), so translation is inhibited. The trans-activating RNA ???key??? (BBa_I714036) can displace the lock and allows translation.
Together, Bxb1 integrase and RDF (BBa_K1039003 and BBa_K1039006) catalyze site-specific recombination between Bxb1 attR and attL sites (Bba_K1039033 and Bba_K1039034)
They will also allow flipping of Bxb1 invertible promoter switches BBa_K1039004 and BBa_K1039004 from RL state to PB state.
Lock and key regulate BXB1 integrase and BRDF expression. BXB1 integrase and BRDF???s expression is blocked by lock. Key expression is controlled by a lacI repressor, upon IPTG addition to the media, key will be expressed, which then unlocks BXB1 integrase and BRDF.
Notes
Bxb1 Integrase (BBa_K1039003) has the following three silent mutations, to remove forbidden restriction sites, base 273 G->A, base 1269 A->T and base 1440 G->ASource
BBa_I714036, BBa_R0011, BBa_J23119 and BBa_Q04121 were obtained from the iGEM 2013 distribution kit. The rest of the biobricks were synthesized by BioBasic Inc.| Access | Instance | Definition |
| public public public public public public public public public public public | BBa_J23119 BBa_Q04121 BBa_J01010 BBa_K1039003 BBa_B0015 BBa_R0011 BBa_I714036 BBa_B0015 BBa_R0011 BBa_J01010 BBa_K1039006 | BBa_J23119 lacI Lock 1 Bxb1-int BBa_B0015 lacI+pL BBa_I714036 BBa_B0015 lacI+pL Lock 1 BBa_K1039006 |