Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: David Caballero
Date created: 2011-09-17 11:00:00
Date modified: 2015-05-08 01:12:30
pUC18R6KT-miniTn7BB-Km
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K510014_sequence (Version 1) |
Description
pUC18R6KT-miniTn7BB-Km is a vehicle vector for the minitransposon miniTn7BB-Km. This transposon is a part of the miniTn7 BioBrick toolkit, a set of plasmids harboring Tn7 derivatives that can be used for integration of BioBricks in single copy at a conserved target (the attTn7 site), in multiple bacterial genomes.The structure of the mini-Tn7BB-Gm transposon is shown in the figure below. Tn7R and Tn7L are the right and left ends of Tn7 transposon, respectively, required for recognition of the transposase machinery and transposition. FRT is the Flp recombinase target site for excision of the resistance once the transposon is inserted in the genome of a living organism. The kanamycin resistance cassette (Km) is flanked by restriction sites in order to facilitate the generation of variants with different antibiotic resistance markers. The BioBrick cloning site (BCS) includes the prefix and suffix restrictions sites used in assembly standard 10, and it is the place where BioBricks are inserted for genome integration.
This plasmid is replicative only in E. coli strains expressing the pir gene thanks to its R6K replication origin. Also it has an origin for transference (oriT) for conjugation purposes. This plasmid can be used to maintain the miniTn7BB-Km transposon in E. coli pir+ strains and may be used for transposition into the genomes of any bacteria without the pir gen, in which it is non-replicative. As a BioBrick cloning vector, pUC18R6KT-miniTn7BB-Km is fully compatible with Assembly Standard 10. The complete transposon is flanked by SfiI restriction sites to facilitate subcloning in other vectors.
Notes
***Source
The pUC18R6KT-mini-Tn7-Km is a derivative of pUC18R6KT-mini-Tn7-Gm (BBa_K510012) by replacing the gentamycin resistance cassette by a kanamycin resistance cassette amplified from pSB4K5 with these primers: tatGCATGCCCATGGattggggctcactcaaagg and tatGCATGCCCATGGtcgacaatgtaactactag.| Sequence Annotation | Location | Component / Role(s) |
| Suffix Tn7-L end SfiI restriction site double terminator (BBa_B0014) Tn7-R end VR (BBa_G00101) primer binding site Ampicillin resistance (bla) R6K replication origin oriT SfiI restriction site E. coli his operon terminator SphI restriction site Flipase recombination sequence (FRT) SfiI restriction site Kanamycin resistance Termiantor NcoI restriction site Flipase recombination sequence (FRT) VF2 (G00100) primer binding site Prefix Terminator | 2,21 182,347 3163,3175 3385,3479 3176,3374 157,176 1714,2574 929,1310 646,905 348,369 22,93 3542,3547 4628,4675 4616,4627 3628,4446 3580,3620 3548,3553 3490,3537 4681,4700 4798,4820 4759,4791 | engineered_region feature/BioBrick feature/misc sequence_feature feature/dna sequence_feature feature/BioBrick engineered_region feature/misc sequence_feature primer_binding_site feature/primer_binding feature/cds CDS feature/dna sequence_feature feature/dna sequence_feature feature/dna sequence_feature stem_loop feature/stem_loop sequence_feature feature/dna feature/dna sequence_feature sequence_feature feature/dna feature/cds CDS feature/stem_loop stem_loop feature/dna sequence_feature sequence_feature feature/dna primer_binding_site feature/primer_binding engineered_region feature/BioBrick stem_loop feature/stem_loop |