Difference between revisions of "Genome Integration"

The parts in the '''miniTn7 BioBrick toolkit'''[http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Overview] are based on the excellent set of miniTn7 derivatives developed by ''Choi et al.'' (2005)[http://www.nature.com/nmeth/journal/v2/n6/abs/nmeth765.html]. All our derivatives contain the Tn7-L and Tn7-R ends, an antibiotic resistance gene flanked by FRTs and the prefix/suffix BioBrick-compatible multi-cloning site. Constructs will be available in derivatives of pUC18-Sfi, (high copy number, replicative in any E. coli strain), or derivatives of pUC18-R6KT (medium copy number, replicative only in E. coli strains expressing the pir gene). Specialized miniTn7 vectors containing promoters for gene expression or reporters for expression measurement will also be constructed. In addition, we will provide a portable attTn7 (Tn7 attachment site) to allow using the miniTn7 BioBrick toolkit in any organism as well as in plasmids.

==We have also submitted these new parts:==

* '''pUC18SfiI-miniTn7BB-Cm''' ([https://parts.igem.org/Part:BBa_K510001 BBa_K510001]): this construct is the result of replacing the gentamycin resistance cassette in pUC18SfiI-miniTn7BB-Gm by a chloramphenicol resistance cassette obtained from pBS1C3 by PCR amplification.

* '''pUC18SfiI-miniTn7BB-Km''' ([https://parts.igem.org/Part:BBa_K510002 BBa_K510002]): replace of the gentamycin resistance cassette of pUC18Sfi-miniTn7-Gm by a kanamycin resistance cassette amplified from pSB4K5.

* '''pUC18R6KT-miniTn7BB-Cm''' ([https://parts.igem.org/Part:BBa_K510013 BBa_K510013]): replace of the gentamycin resistance cassette in pUC18R6KT-miniTn7BB-Gm by a chloramphenicol resistance cassette obtained from pBS1C3 by PCR amplification with these primers

*''' pUC18R6KT-miniTn7BB-Gm-RBS+RFP''' ([https://parts.igem.org/Part:BBa_K510015 BBa_K510015]): This plasmid can be used for promoter characterization purposes inserting the promoter using the prefix restriction sites. Also the characterization can be performed in single copy by integrating the device in the genome of the working organism.

* '''pUC18R6KT-miniTn7BB-Gm-pBad/araC''' ([https://parts.igem.org/Part:BBa_K510016 BBa_K510016]): This plasmid can be used as inducible expression vector in single copy by integrating the device in the genome of the working organism.

* '''pUC18R6KT-miniTn7BB-Gm-tetR''' ([https://parts.igem.org/Part:BBa_K510017 BBa_K510017]): This plasmid can be used as inducible expression system in single copy by integrating the device in the genome of the working organism.

*''' pUC18R6KT-miniTn7BB-Gm-lacZ+GFP''' ([https://parts.igem.org/Part:BBa_K510018 BBa_K510018]): This plasmid can be used for promoter characterization purposes inserting the promoter using the prefix restriction sites. Also the characterization can be performed in single copy by integrating the device in the genome of the working organism.

* '''pUC18R6KT-miniTn7BB-Gm-RFP''' ([https://parts.igem.org/Part:BBa_K510020 BBa_K510020]): This plasmid can be used for visualization purposes or to brand a strain because of its insertion in the genome of the working organism and split the antibiotic resistance cassette with the temporary expression of Flp recombinase.

* '''pUC18R6KT-miniTn7BB-Gm-Lux''' ([https://parts.igem.org/Part:BBa_K510021 BBa_K510021]): This plasmid can be used for visualization purposes or to brand a strain because of its insertion in the genome of the working organism.