Then well yes, i think you set the positive control wrong. Pdf engineering a minimal cloning vector from a puc18 plasmid. One such benefit is the ability to produce large quantities of biological materials that. A plasmid is a circular dsdna molecule a few hundred or thousand base pairs in circumference.
This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Purified by chromatography using proprietary patented technology. These cloning vectors contain a multiple cloning site at the lacz region that enables recombinant plamids to be verified via bluewhite colony screening using agar plates containing iptg and xgal. The bacterium of choice is a modified escherichia coli strain which allows it uptake of the plasmid puc18. Staphylococcus aureus kanamycin nucleotidyltransferase genomic dna. Use our dna sequences and maps tool to view the sequence files used to produce. This is a free resource for the scientific community that is compiled by addgene.
Likewise, the atrich genome of lactobacillus helveticus 65% at was cloned. The molecule is a small doublestranded circle, 2686 base pairs in length, and has a high copy number. The pmb1 replicon rep responsible for the replication of plasmid source plasmid. The latter one, pucp18, has a broad host range origin that was cloned into the puc18 plasmid, so it will replicate in many gram negative bacteria. You can add anything we like, change the details and make our. The coordinates refer to the position of the first nucleotide in each. We offer a huge selection of easytohandle tools for efficient gene expression in different microorganisms like bacillus megaterium, bacillus. The cut sites for some restriction enzyme are indicated on the plasmid. Xx rn 1 rp 12686 rc mmp18 from mmp8 rc mmp19 from mmp9 rc puc18 from puc8 rc puc19 from puc9 ra yanischperron c. The nucleotide sequence files available below are those used to produce the plasmid vector, viral and bacteriophage maps contained in new england biolabs catalog as well as the tables containing the locations of sites. The map, notes, and annotations on this page and in the sequence map file are ed material. Plasmid transformation using puc18 on e coli using.
Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circular map. A cloning vector is a small piece of dna that can be stably maintained in an organism, and into which a foreign dna fragment can be inserted for cloning purposes. The difference is that pgem3z has two additional, short pieces of dna, each of which acts as the recognition site for attachment of an rna polymerase enzyme. The artificial plasmid puc18 has been genetically engineered to include 1 a gene for antibiotic resistance to ampicillin amp r, and 2 a gene and its promoter for the enzyme betagalactosidase lacz. Restriction site coordinates refer to the position of the 5. United states city plans vector street maps and full us maps in adobe illustrator, pdf and other vector formats. Free vector maps in ai, eps, pdf, svg, jpg and png formats. Thermo scientific puc19 vector is a small, high copy number, e.
This material is available to academics and nonprofits only. To see this sequence with restriction sites, features. The puc18 puc19 sequence is stored as a pdf file on the genaxxon webpage. The vector length is 2686 bp and is isolated from e. It would be hard to get ligated a circular plasmid puc19 into the topo vector, so i would not be surprised that you had no colonies.
You should also get these plasmid dna restriction map, puc18 vector map and plasmid vector map, its magical vector map. Us city maps vector urban plans in the adobe illustrator. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Plasmid is a more descriptive as it is a type of vector. For publishing, design, printing, media, publications, arts, projects, presentations, for architects, designers and builders, business, logistics. I did go through the fermentas website, and found the restriction map of puc18 i have attached it for clarity but couldnt decipher it. Article engineering a minimal cloning vector from a puc18. The archive is devided into 7 main categories which consist of subcategories countries of the continent. Mobitec is leading in the development of unique and cutting edge vector systems for recombinant protein expression. Naturallyoccurring plasmids are viruses of bacteria.
The puc18 and puc19 plasmids enable successful cloning of large dna fragments larger than those cloned with a m mp18 rf phage vector. You will see something best in puc19 vector sequence primer, plasmid restriction map and puc19 plasmid map, it can give ideas to make our own graphic work. In the first article in this series, we talked about how origins of replication ori control plasmid replication and copy number. It contains identical multiple cloning site mcs as puc18 vector except that it is arranged in opposite orientation. Restriction map of puc18 oct232005 i just was curious about the size of the fragments resulting from the digestion of the vector puc18 with hindiii first and then ecori. Overview of primers used in the iterative shrinking of puc18 into picoz. Today i would like to introduce you to puc18, a plasmid most noted for its high copy number. Ligation reactions using this buffer may be incubated for 1 hour at room temperature. The in silico cloning, graphical vector map generation and sequence analyses was done in ugene. To learn about this, we focused on the pbr22 ori and the role of rop protein in controlling copy number within pbr22 and other members of the cole1 family. The cloning vector may be dna taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. What is the difference between puc18 and puc19 answers.