We have added a new section for the "Plasmid backbones for advanced users" of the Parts' Registry: Genome Integration plasmids, where you could find all the plasmids of the miniTn7 BioBrick tool kit. Enjoy!

Data for our characterized new parts

1. pUC18Sfi-miniTn7BB-Gm plasmid (BBa_K510000): This plasmid harbors the basic miniTn7 construct and may be used for integration of BioBrick parts and devices in single copy into the genomes of non-enteric bacteria, in which it is non-replicative, at the conserved attTn7 site.

2. pUC18R6KT-miniTn7BB-Gm plasmid (BBa_K510012): This plasmid harbors the basic miniTn7 construct and may be used for transposition of BioBrick parts and devices into the genome of bacteria not expressing the pir gene (including Enterobacteria), thanks to is R6K replication origin.

3. attTn7 or Tn7 insertion site (BBa_K510022): Recognition and insertion site for the Tn7 transposon and its synthetic derivatives. Developed to promote target-specific Tn7 transposition in hosts not bearing a functional Tn7 insertion site.

Moreover, we have constructed these new parts:

1. pUC18SfiI-miniTn7BB-Km (BBa_K510002): This construct is the result of replacing the gentamycin resistance cassette of pUC18Sfi-miniTn7-Gm by a kanamycin resistance cassette amplified from pSB4K5.

2. pUC18Sfi-miniTn7BB-Gm-RBS+RFP (BBa_K510037): This plasmid can be used for promoter characterization purposes inserting the promoter using the prefix restriction sites, measuring the RFP fluorescence. Also the characterization can be performed in single copy by integrating the device in the genome of the working organism using the transposase machinery of the Tn7 transposon.

3. pUC18Sfi-miniTn7BB-Gm-lacZ+GFP(BBa_K510040): This plasmid can be used for promoter characterization purposes inserting the promoter using the prefix restriction sites, measuring LacZ-alpha and/or GFP-AAV expresion. Also the characterization can be performed in single copy by integrating the device in the genome of the working organism.

4. pUC18Sfi-miniTn7BB-Gm-RFP (BBa_K510041): 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. Also it is a useful plasmid for clonning BioBricks into the miniTn7 module by removing the RFP cassette. Negative clones of a transformation would be in red colour.

5. pUC18Sfi-miniTn7BB-Gm-bistable_switch (BBa_K510043): This plasmid allows the integration of a transcriptional flip-flop into bacterial chromosomes using the pUC18Sfi-miniTn7BB-Gm (BBa_K51000) characteristics. Single copy may improve the function of regulatory circuits, as bistable systems.

6. pUC18R6KT-miniTn7BB-Gm-bistable_switch (BBa_K510044): This plasmid allows the integration of a transcriptional flip-flop into bacterial chromosomes using the pUC18R6KT-miniTn7BB-Gm (BBa_K510012)characteristics. Single copy may improve the function of regulatory circuits, as bistable systems.

7. pUC18Sfi-miniTn7BB-Gm-improved_flipflop_(module I) (BBa_K510045): This plasmid allows the integration of the improved flip-flop (module I) (part: BBa_K51019) into bacterial chromosomes using the pUC18Sfi-miniTn7BB-Gm (BBa_K51000) characteristics. Single copy may improve the function of regulatory circuits, as bistable systems.

8. pUC18R6KT-miniTn7BB-Gm-improved_flipflop_(module I) (BBa_K510046): This plasmid allows the integration of the improved flip-flop (module I) (part: BBa_K51019) into bacterial chromosomes using the pUC18R6KT-miniTn7BB-Gm (BBa_K51012) characteristics. Single copy may improve the function of regulatory circuits, as bistable systems.

We have also submitted these new parts:

1. pUC18SfiI-miniTn7BB-Cm (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.

2. pUC18R6KT-miniTn7BB-Cm (BBa_K510013). This construct is the result of replacing the gentamycin resistance cassette in pUC18R6KT-miniTn7BB-Gm by a chloramphenicol resistance cassette obtained from pBS1C3 by PCR amplification with these primers

3. pUC18R6KT-miniTn7BB-Km (BBa_K510014). This construct is the result of replacing the gentamycin resistance cassette in pUC18R6KT-miniTn7BB-Gm by a kanamycin resistance cassette obtained from pBS4K5.

4. pUC18R6KT-miniTn7BB-Gm-RBS+RFP (BBa_K510015). MiniTn7-based reporter for promoter characterization. Promoter insertion at prefix restriction sites results in a transcriptional fusion to RFP. Characterization can be performed in single copy by integrating the miniTn7 element in the genome of the appropriate organism.

5. pUC18R6KT-miniTn7BB-Gm-pBad/araC (BBa_K510016). MiniTn7-based expression vector. Insertion of genes at suffix restriction sites allows arabinose-inducible transcription. Characterization can be performed in single copy by integrating the miniTn7 element in the genome of the appropriate organism.

6. pUC18R6KT-miniTn7BB-Gm-tetR (BBa_K510017). MiniTn7-based expression vector. Insertion of genes at suffix restriction sites allows oxytetracycline-inducible transcription. Characterization can be performed in single copy by integrating the miniTn7 element in the genome of the appropriate organism.

7. pUC18R6KT-miniTn7BB-Gm-lacZ+GFP (BBa_K510018). MiniTn7-based reporter for promoter characterization. Promoter insertion at prefix restriction sites results in a transcriptional fusion to lacZ alpha fragment and GFP. Characterization can be performed in single copy by integrating the miniTn7 element in the genome of the appropriate organism.

8. pUC18R6KT-miniTn7BB-Gm-RFP (BBa_K510020). MiniTn7-based vector for fluorescent labeling of bacterial strains. Single copy labeling with constitutively-produced RFP is achieved by integrating the miniTn7 element in the genome of the appropriate organism.

9. pUC18R6KT-miniTn7BB-Gm-Lux (BBa_K510021): MiniTn7-based vector for biomoluminescent labeling of bacterial strains. Single copy labeling with LuxAB is achieved by integrating the miniTn7 element in the genome of the appropriate organism.

10. pUC18Sfi-miniTn7BB-Gm-tetR (BBa_K510039): This plasmid can be used as inducible expression system in single copy by integrating the device in the genome of the working organism using the pUC18Sfi-miniTn7BB-Gm characteristics.

11. pUC18Sfi-miniTn7BB-Gm-Lux (BBa_K510042): 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.

12. pUC18Sfi-miniTn7BB-Gm-attTn7 (BBa_K510047): This plasmid allows the integration of BioBricks into microbial chromosomes, using prefix or suffix cloning sites, without removing the attachment site of the Tn7 transposon (attTn7). This fact makes posible to integrate another BioBrick into the chromosome of a strain in which miniTn7BB-Gm-attTn7 transposition has occurred previously. Based on pUC18Sfi-minitn7BB-Gm and attTn7.

13. pUC18R6KT-miniTn7BB-Gm-attTn7 (BBa_K510048): This plasmid allows the integration of BioBricks into microbial chromosomes, using prefix or suffix cloning sites, without removing the attachment site of the Tn7 transposon (attTn7). This fact makes posible to integrate another BioBrick into the chromosome of a strain in which miniTn7BB-Gm-attTn7 transposition has occurred previously. Based on pUC18R6KT-miniTn7BB-Gm and attTn7.