Puc19 vector map pdf

To see this sequence with restriction sites, features. Vector map of puc19 puc19 is one of a series of plasmid cloning vectors created by joachim messing and coworkers. The designation puc is derived from the classical p prefix denoting plasmid and the abbreviation for the university of california, where early work on the plasmid series had been conducted. It is a circular double stranded dna and has 2686 base pairs. Left plate serves as the control, with vector backbone only, right plate contains pcr insert. All restriction sites specified in the vector map were checked by sequencing.

Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. 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. Thermo scientific puc19 vector is a small, high copy number, e. The multiple cloning site mcs is within the bgalactosidase gene. Product information keywords puc19 dna, molecular cloning products, thermo scientific. If you wish to digest the vector with these enzymes, you will need to transform the vector into a dam host and make fresh dna. A, a map of the plasmid pgfpuv, which was purchased from clontech. Purified by chromatography using proprietary patented technology. A desktop resource, is to curate a onestop reference guide for plasmids. It contains identical multiple cloning site mcs as puc18 vector except that it is arranged in opposite orientation. Additional enzyme cut sites are indicated on this map. Hiv1 vpr gene segment was amplified from pbsxcypyvprxcthy with polymerase chain reaction pcr, and then cloned into pgemt vector and the cloning vector pgemt vpr was obtained.

It contains the pmb1 origin of replication from pbr322, but it lacks the rop gene and carries a point mutation in the rnaii transcript g 2975 in pbr322 to a 8 in puc19. Restriction map and multiple cloning site mcs of pegfpc1. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the vector map. This means it is a circular piece of dna 2686 base pairs in length. Dna fragments into puc19 vector to study the ligation. This vector can be used in combination with pacycduet1 cat. L097 feature lacz origin bla apr coordinates 469146 1455867 24861626 enzymes with unique.

Each order of pet dna also includes an induction control strain supplied as a glycerol stock. Vector id construct host vector stock selection notes. Expression of target dna is enabled by the presence of a. The xbai site in the puc19 mcs has been replaced with a spei site. The vector encodes two multiple cloning sites, each of which is preceded by a t7 promoter, lac operator and ribosome binding sites.

The xba i and bcl i sites are methylated in the dna provided by bd biosciences clontech. The xbai site in the puc19 mcs has been replaced with a spei site these sites produce compatible cohesive ends when cleaved. The vector also carries the rsf1030derived rsf replicon, laci gene and kanamycin resistance gene knr. It can be downloaded from the detaisl view of the puc18puc19 product description. If you then cut that piece into two equallength pieces, you would have two pieces of dna each 43 base pairs long. The puc18 and puc19 plasmids enable successful cloning of large dna fragments larger than those cloned with a m mp18 rf phage vector.

The map, notes, and annotations on this page and in the sequencemap file are ed material. The order of the restriction sites in this vector are almost the same as in puc19 however the sequence is not. Locus puc19 2686 bp dna circular syn 27sep2006 definition classical cloning vector accession puc19 keywords. V33202 puc19 vector dna 25 g riba25 rnase a, 90 umg kunitz 25 mg a141425gmag ampicillin, sodium salt 25 g i121gag iptg isopropylbetadthiogalactoside 1 g x10155gag xgal 5 g 04004g mobitec agarose le 500 g contact and support mobitec gmbh lotzestrasse 22a d37083 goettingen germany.

This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Location of sites of all neb restriction enzymes can be found on the neb web site choose technical reference dna sequences and maps. Novagen ordering 8005267319 technical support 8002070144 prsfduet1 vector tb391 0903 prsfduet1 sequence landmarks t7 promoter1 35823598 t7 transcription start1 1 histag coding sequence 83100 multiple cloning sites1 nco ia. Reference 1 bases 1 to 2686 authors self journal unpublished.

The pgem t and pgem t easy vector systems have been optimized using a 1. Psfcmvpuc19 cmv puc19 mcs plasmid plasmid vector for. This allows pcr inserts to ligate efficiently with the vector. The molecule is a small doublestranded circle, 2686 base pairs in length, and has a high copy number. Cloning vector puc18 mobitec molecular biotechnology. Terms and conditions 20042018 harvard medical school plasmid was created and is maintained by the dfhcc dna resource core at harvard medical schooldfhcc dna resource. The position of the multi cloning site mcs, which is inserted in frameinto the laczgene, enables a bluewhite selection of insert containing plasmid dna by complemen tation. For example, telomeric repeats and other atrich fragments from pneumocystis carinii 60% to 65% at were stable in a low copy, transcriptionfree vector but were unstable in puc19. The pbart construct was confirmed by restriction mapping and sequencing. The molecule is a small doublestranded circle, 2686 base.

This is a free resource for the scientific community that is compiled by addgene this page is informational only this vector is not available from addgene please contact the manufacturer for further details. The puc19 plasmid 2,686 bp confers ampicillin resistance and complement defects in. The puc18puc19 sequence is stored as a pdffile on the genaxxon webpage it can be downloaded from the. Additional restriction sites that can be used for subcloning are shown in red. The final chimeric protein carries gfp at the nterminal end, a his 6 affinity tag flanked by flexible glycine fig. Terms and conditions 20042018 harvard medical school plasmid was created and is maintained by the dfhcc dna resource core at harvard medical schooldfhcc dna resource core at. Discrepancies between sequencing results obtained by addgene and the original sequence provided by the depositor may be present. Gene amplification by pcr and subcloning into a gfpa fusion. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circular map. All plasmid maps were created by the shareware program macplasmap v1. Description puc19 is a standard highcopy cloning vector for li recombinants.

It has been constructed using the ampicillin resistance gene and the pmb1 origin of replication from pbr322. To see this sequence with restriction sites, features, and translations, please. Construction of an enlarged puc19 vector with a rop gene designed. Product information puc19 dna features applications storage. 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. If initial experiments with your pcr product are suboptimal, ratio optimization may be necessary. Description puc19 is a small, high copy cloning vector for replication in e.

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. 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. This is a free resource for the scientific community that is compiled by addgene. Find msds or sds, a coa, data sheets and more information. The linearized vector supplied in this kit has single, overhanging 3. Creating a map of puc19 puc19 is a plasmid that has 2686 base pairs. The vector contains two multiple cloning sites mcs, each of which is preceded by a t7lac promoter and ribosome binding site rbs. The puc19 linearized vector is provided as a part of infusion cloning kits, and is intended to serve as a control for infusion cloning. The mfwd and mrev primer binding sties for pcr amplification and sequencing of inserts are indicated.

Product information puc19 dna features applications. Highlights purified by chromatography using proprietary patented technology more than. Creating a map of puc19 puc19 is a plasmid that ha. Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources.

Then well yes, i think you set the positive control wrong. We take several day to collect these fresh vector map pictures from best creator. Overall cloning scheme to produce a gfphis 6serpin construct. A cloning vector is a genome that can accept the target dna and increase the number of copies through its own autonomous replication. This resource is designed to educate all levels of scientists and plasmid lovers and serves as an introduction to plasmids, allowing you to spend. The vector length is 2686 bp and is isolated from e. The molecule is a small doublestranded circle, 2686 base pairs in length. The map, notes, and annotations on this page and in the sequence map file are ed material. Sequence provided by depositing laboratory may be theoreticalpredicted or based on sangerngs sequencing results. Ifit was cut so that it was a linear piece of dna then it would still be 2686 base pains long. The puc18puc19 sequence is stored as a pdf file on the genaxxon webpage.